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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Форма поиска

Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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Применить Всего найдено 8597. Отображено 102.
01-12-1997 дата публикации

Номер: KR19977007277A
Автор:
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26-04-1989 дата публикации

Номер: KR19897000676A
Автор:
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10-04-2015 дата публикации

СПОСОБ КЛОНИРОВАНИЯ ГИБРИДОМ

Номер: RU0000151761U1

Способ клонирования гибридом, заключающийся в визуально контролируемом заборе клеток и перенесении их в питательную среду для дальнейшего культивирования, отличающийся тем, что после слияния и выбора объектов клонирования под визуальным контролем прицельно отбирают гибридомные клетки четко отграниченного клона пипеточным микродозатором с насадкой, имеющей тонкую длинную носовую часть, после чего клетки переносят в свежую питательную среду для дальнейшего культивирования. Ц 151761 ко РОССИЙСКАЯ ФЕДЕРАЦИЯ ВО“ 151 761” 91 ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ИЗВЕЩЕНИЯ К ПАТЕНТУ НА ПОЛЕЗНУЮ МОДЕЛЬ ММ9К Досрочное прекращение действия патента из-за неуплаты в установленный срок пошлины за поддержание патента в силе Дата прекращения действия патента: 01.10.2018 Дата внесения записи в Государственный реестр: 04.07.2019 Дата публикации и номер бюллетеня: 04.07.2019 Бюл. №19 Стр.: 1 па эра ЕП

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10-01-2017 дата публикации

УСТРОЙСТВО ДЛЯ ГИДРОЛИЗА ВОДНЫХ РАСТВОРОВ БЕЛКОВ

Номер: RU0000167833U1

Использование: биотехнология, в пищевой промышленности, медицине и сельском хозяйстве. Сущность решения: в обогреваемый водой ферментер 1 через штуцеры 3 и 4 подают соответственно суспензию иммобилизированного фермента и водный раствор белка. Отделение полученного гидролизата белка от общей реакционной массы происходит на ультрафильтрационной мембране 8, расположенной в днище ферментера, через штуцер 6. Подача сжатого воздуха в ферментер 1 осуществляется снизу через вращающуюся камеру барботера 12, плотно прилегающую верхним открытым краем к дренажной пластине 9. В крышке ферментера 1 установлен штуцер 7 с вентилем для регулирования давления в ферментере 1. 1 ил. РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 167 833 U1 (51) МПК C12M 1/40 (2006.01) C12N 11/02 (2006.01) C12P 21/06 (2006.01) C07K 1/12 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ПОЛЕЗНОЙ МОДЕЛИ К ПАТЕНТУ (21)(22) Заявка: 2016126447, 01.07.2016 (24) Дата начала отсчета срока действия патента: 01.07.2016 Дата регистрации: (73) Патентообладатель(и): Федеральное государственное бюджетное образовательное учреждение высшего образования "Московский государственный университет леса" (RU) Приоритет(ы): (22) Дата подачи заявки: 01.07.2016 (45) Опубликовано: 10.01.2017 Бюл. № 1 20150159189 A1, 11.06.2015. R U 1 6 7 8 3 3 (54) УСТРОЙСТВО ДЛЯ ГИДРОЛИЗА ВОДНЫХ РАСТВОРОВ БЕЛКОВ (57) Реферат: Использование: биотехнология, в пищевой расположенной в днище ферментера, через промышленности, медицине и сельском хозяйстве. штуцер 6. Подача сжатого воздуха в ферментер Сущность решения: в обогреваемый водой 1 осуществляется снизу через вращающуюся ферментер 1 через штуцеры 3 и 4 подают камеру барботера 12, плотно прилегающую соответственно суспензию иммобилизированного верхним открытым краем к дренажной пластине фермента и водный раствор белка. Отделение 9. В крышке ферментера 1 установлен штуцер 7 полученного гидролизата белка от общей с вентилем для регулирования давления в реакционной массы ...

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01-02-2018 дата публикации

Устройство для гидролиза водных растворов биополимеров

Номер: RU0000176889U1

Использование: биотехнология, медицина, пищевая промышленность, сельское хозяйство. Сущность решения: в стеклянный или металлический вращающийся реактор через вводной штуцер в верхний слой роторного реактора подают под давлением водный раствор белков или полисахаридов или липидов и водную суспензию фермента или кислоты. Степень гидролиза биополимера контролируют с помощью отбора проб. Для отделения полученного гидролизата от реакционной массы используют штуцер для удаления продукта из пристеночной части пленочного слоя. 1 ил. И 1 176889 ко РОССИЙСКАЯ ФЕДЕРАЦИЯ 7 ВУ"” 176 889” 91 ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ИЗВЕЩЕНИЯ К ПАТЕНТУ НА ПОЛЕЗНУЮ МОДЕЛЬ МЕЭК Восстановление действия патента Дата, с которой действие патента восстановлено: 16.01.2020 Дата внесения записи в Государственный реестр: 16.01.2020 Дата публикации и номер бюллетеня: 16.01.2020 Бюл. №2 Стр.: 1 па 638911 ЕП

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19-08-2020 дата публикации

УСТРОЙСТВО ДЛЯ СИНТЕЗА БЕЛКОВ В БЕСКЛЕТОЧНОЙ СИСТЕМЕ

Номер: RU0000199182U1

Полезная модель относится к области молекулярной биологии и биотехнологии. Устройство для синтеза белков в бесклеточной системе содержит реакционную ячейку с первым и вторым пористыми барьерами, образующими объем с реакционной смесью. Насосы обеспечивают подачу раствора питающей смеси в реакционный объем и отвод раствора продуктов синтеза. Устройство дополнительно содержит внешнюю по отношению к реакционной ячейке жидкостную коммуникацию, которая соединяет вход и выход реакционного объема с помощью последовательно соединенных первого и второго крана и насоса. Данная жидкостная коммуникация позволяет обеспечить частичную или полную смену реакционной смеси в ячейке и продлить эффективный синтез целевых белков. 6 з.п. ф-лы, 1 ил. РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (51) МПК C12M 1/12 C12M 1/14 C12P 21/00 C07K 1/02 (11) (13) 199 182 U1 (2006.01) (2006.01) (2006.01) (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ПОЛЕЗНОЙ МОДЕЛИ К ПАТЕНТУ (52) СПК C12M 1/12 (2020.02); C12M 1/14 (2020.02); C12P 21/00 (2020.02); C07K 1/02 (2020.02) (21)(22) Заявка: 2019140794, 10.12.2019 (24) Дата начала отсчета срока действия патента: 19.08.2020 Приоритет(ы): (22) Дата подачи заявки: 10.12.2019 (45) Опубликовано: 19.08.2020 Бюл. № 23 1 9 9 1 8 2 R U (54) УСТРОЙСТВО ДЛЯ СИНТЕЗА БЕЛКОВ В БЕСКЛЕТОЧНОЙ СИСТЕМЕ (57) Реферат: Полезная модель относится к области внешнюю по отношению к реакционной ячейке молекулярной биологии и биотехнологии. жидкостную коммуникацию, которая соединяет Устройство для синтеза белков в бесклеточной вход и выход реакционного объема с помощью системе содержит реакционную ячейку с первым последовательно соединенных первого и второго и вторым пористыми барьерами, образующими крана и насоса. Данная жидкостная объем с реакционной смесью. Насосы коммуникация позволяет обеспечить частичную обеспечивают подачу раствора питающей смеси или полную смену реакционной смеси в ячейке и в реакционный объем и отвод раствора продуктов продлить эффективный синтез ...

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09-02-2012 дата публикации

Microorganism and method for producing canthaxanthin

Номер: US20120034649A1
Автор: Teruhiko Ide, Toru Tanaka
Принадлежит: Tosoh Corp

A carotenoid producing bacterium belonging to the genus Paracoccus that selectively produces canthaxanthin so that the amount thereof is not less than 90 percent by weight of the total amount of produced carotenoids including β-carotene, β-cryptoxanthin, echinenone, canthaxanthin, 3-hydroxyechinenone, 3′-hydroxyechinenone, zeaxanthin, phoenicoxanthin, adonixanthin, and astaxanthin. A method for producing canthaxanthin by culturing the above bacterium, and then collecting carotenoids from bacterial cells or a culture solution after the culturing.

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23-02-2012 дата публикации

Enzyme-Based Fed-Batch Technique In Liquid Cultures

Номер: US20120045836A1
Принадлежит: BIOSILTA OY

The present invention is generally in the field of continuous and high-cell-density cultivation in laboratory- or large-scale liquid shaken cultures. More particularly it relates to a method of enzyme-based fed-batch (EnBase) for liquid microbial prokaryotic or eukaryotic cell cultivation having the possibility to manipulate the growth rate of the cultured organisms by a controlled enzymatic release of the growth-limiting substrate-monomer from substrate-polymers or substrate-oligomers.

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24-05-2012 дата публикации

Process for production of useful substance, and surfactant for use in the process

Номер: US20120129220A1
Принадлежит: Sanyo Chemical Industries Ltd

The present invention is aimed to provide a method for industrially producing a useful substance (protein etc.) utilizing a bacterium such as Escherichia coli in the presence of a surfactant and a surfactant to be used in this method for producing a useful substance. The present invention is a method for producing a useful substance into a culture solution by secretion by a bacterium in the culture solution, wherein the culture solution contains a surfactant (A), and a dried cell density based on the volume of the culture solution is 1.5 to 500 g/L. More preferably, the present invention is a method for producing a useful substance wherein the surfactant (A) is at least one agent selected from the group consisting of an amphoteric surfactant, an anionic surfactant, and a nonionic surfactant having an HLB value of 0 to 13.

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31-05-2012 дата публикации

Method for Producing Nucleases of a Gram Negative Bacterium While Using a Gram Positive Expression Host

Номер: US20120135498A1
Принадлежит: C Lecta GmbH

A method for producing a nuclease of a gram negative bacterium or a nuclease preparation containing a nuclease of a gram negative bacterium including expression of the nuclease in a gram positive bacterium and subsequent secretion of the nuclease, as well as a nuclease or a nuclease preparation that can be obtained by this method.

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17-01-2013 дата публикации

Methods and compositions for enhanced expression and secretion of proteins

Номер: US20130017574A1
Автор: Xiaowu Liu, Zhuying Wang

Optimized signal peptide coding sequences for enhanced expression and secretion of protein from a cell and related compositions and methods are described. The optimized signal peptide coding sequence encodes an mRNA that contains at least one hairpin structure immediately downstream of the initiation codon. Methods for obtaining the optimized signal peptide coding sequences and methods for enhanced expression and secretion of proteins using the optimized signal peptide coding sequences are also described.

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14-03-2013 дата публикации

Synthesis of Site Specifically-Linked Ubiquitin

Номер: US20130065272A1
Принадлежит: Medical Research Council

The invention relates to a method of modifying a specific lysine residue in a polypeptide comprising at least two lysine residues, said method comprising (a) providing a polypeptide comprising a target lysine residue protected by a first protecting group, and at least one further lysine residue; (b) treating the polypeptide to protect said further lysine residue(s), wherein the protecting group for said further lysine residues is different to the protecting group for the target lysine residue; (c) selectively deprotecting the target lysine residue; and (d) modifying the deprotected lysine residue of (c). 1. A method of modifying a specific lysine residue in a polypeptide comprising at least two lysine residues , said method comprising:a. providing a polypeptide comprising a target lysine residue protected by a first protecting group, and af least one further lysine residue;b. treating the polypeptide to protect said further lysine residue(s), wherein the protecting group for said further lysine residues is different to the protecting group for the target lysine residue;c. selectively deprotecting the target lysine residue; andd. modifying the deprotected lysine residue of (c).2. A method according to wherein producing the polypeptide comprisesa. providing a nucleic acid encoding the polypeptide which nucleic acid comprises an orthogonal codon encoding the target lysine;b. translating said nucleic acid in the presence of an orthogonal tRNA synthetase/tRNA pair capable of recognising said orthogonal codon and incorporating said target lysine residue protected by a first protecting group into the polypeptide chain.3. A method according to wherein said orthogonal codon comprises TAG claim 2 , said tRNA comprises MDtRNACUA and said tRNA synthetase comprises MbPylRS.4. A method according to claim 1 , wherein the target lysine residue protected by a first protecting group is Nε-(1-butyloxycarbonyl)-L-lysine.5. A method according to claim 1 , wherein the protecting group ...

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18-04-2013 дата публикации

Method of Supplementing Culture Media to Prevent Undesirable Amino Acid Substitutions

Номер: US20130096283A1
Принадлежит: BIOGEN IDEC MA INC.

The present invention relates to methods of reducing substitution of amino acids during the production of polypeptides of interest in mammalian cells. By varying the concentration of amino acids in the culture medium, heterogeneity of the polypeptide's amino acid sequence is decreased, leading to higher yields of functional protein. 1. A method for reducing substitution of a first amino acid by a second amino acid during translation of a polypeptide of interest in an eukaryotic cell , comprising:(a) culturing the cell in growth media that is supplemented with the first amino acid, or a metabolic precursor thereof, in an amount sufficient to reduce amino acid substitution; and/or(b) culturing the cell in growth media in which the amount of the second amino acid is reduced.2. The method of claim 1 , wherein the first amino acid is an essential amino acid selected from the group consisting of: arginine claim 1 , histidine claim 1 , isoleucine claim 1 , leucine claim 1 , lysine claim 1 , methionine claim 1 , phenylalanine claim 1 , threonine claim 1 , tryptophan and valine.3. The method of claim 1 , wherein the first amino acid is a non-essential amino acid present in limiting concentrations.4. (canceled)5. The method of claim 1 , wherein the first amino acid is glutamine or asparagine.67.-. (canceled)8. The method of claim 1 , wherein the second amino acid is serine.9. (canceled)10. The method of claim 1 , wherein the first amino acid is provided during the growth or production phase.1112.-. (canceled)13. The method of claim 1 , wherein the first amino acid is added prior to depletion.14. The method of claim 1 , wherein the first amino acid is added at a concentration of greater than about 0.1 mM.1519.-. (canceled)20. The method of claim 1 , wherein the mammalian cell is selected from the group consisting of: chinese hamster ovary (CHO) claim 1 , monkey kidney CV1 claim 1 , monkey kidney COS claim 1 , human lens epithelium claim 1 , human embryonic kidney claim 1 , ...

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16-05-2013 дата публикации

MODIFIED SAK GENE FOR THE PRODUCTION OF RECOMBINANT PROTEINS

Номер: US20130122542A1
Принадлежит: Lupin Limited

The present invention relates to modified SAK gene having amino acid SEQ ID 2. The present invention further relates to process for cloning and expressing modified SAK gene fusion protein which imparts improved stability to the heterologous protein of interest. Further the invention relates to process of purification of recombinant heterologous proteins from bacterial inclusion bodies using modified SAK. 1. A fusion DNA comprising a first DNA encoding a modified SAK protein comprising nucleic acid sequence of SEQ ID NO. 1 and a second DNA fused in frame encoding the heterologous protein of interest.2. The fusion DNA as claimed in claim 1 , comprising amino acid sequence of SEQ ID NO. 2.3. The fusion DNA as claimed in claim 1 , wherein the heterologous protein of interest is selected from the group comprising parathyroid hormone (1-34) claim 1 , parathyroid hormone (1-84) claim 1 , reteplase claim 1 , interferon claim 1 , IL-2 claim 1 , IL-3 claim 1 , IL-4 claim 1 , IL-5 claim 1 , IL-6 claim 1 , IL-11 claim 1 , GCSF claim 1 , epidermal growth factor and platelet derived growth factor.4. The fusion DNA as claimed in claim 1 , wherein the heterologous protein of interest is parathyroid hormone (1-34) having SEQ ID NO. 3.5. The fusion DNA as claimed in claim 1 , wherein the heterologous protein of interest is parathyroid hormone (1-34) having SEQ ID NO. 4.6. The fusion DNA as claimed in claim 1 , wherein the heterologous protein of interest is IL-11 having SEQ ID NO. 5.7. The fusion DNA as claimed in claim 1 , wherein the heterologous protein of interest is IL-11 having SEQ ID NO. 6.8. The fusion DNA as claimed in claim 1 , wherein the modified SAK protein carries an enterokinase site inside the SAK gene.9E. coli. A process for the preparation of heterologous protein of interest in comprising the steps of:a) preparing a fusion DNA comprising a first DNA encoding modified SAK protein and a second DNA fused in the frame encoding the heterologous protein of interest,b) ...

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23-05-2013 дата публикации

METHODS AND STRAINS FOR THE PRODUCTION OF SARCINAXANTHIN AND DERIVATIVES THEREOF

Номер: US20130130312A1
Принадлежит: Promar AS

The present invention relates to a new strain of , named Otnes7, which is superior to known strains in its ability to synthesise the carotenoid sarcinaxanthin and a method of producing sarcinaxanthin or a derivative thereof, said method comprising introducing into and expressing in a host cell one or more nucleic acid molecules encoding an activity in the sarcinaxanthin biosynthetic pathway. 1. A method of producing sarcinaxanthin or a derivative thereof , said method comprising introducing into and expressing in a host cell one or more nucleic acid molecules encoding an activity in the sarcinaxanthin biosynthetic pathway , wherein said one or more nucleic acid molecules comprise:(i) a nucleotide sequence as set forth in SEQ ID NO: 37 or a part thereof;(ii) a nucleotide sequence with at least 90% sequence identity to SEQ ID NO: 37, or a part thereof; or(iii) a nucleotide sequence complementary to (i) or (ii).2. The method of claim 1 , wherein said one or more nucleic acid molecules comprise:(i) a nucleotide sequence as set forth in SEQ ID NO: 26 or a part thereof;(ii) a nucleotide sequence with at least 90% sequence identity to SEQ ID NO: 26, or a part thereof; or(iii) a nucleotide sequence complementary to (i) or (ii).3. The method of claim 1 , wherein said one or more nucleic acid molecules encode the sarcinaxanthin biosynthetic pathway.4. The method of claim 1 , further comprising the step of isolating the sarcinaxanthin or derivative thereof from the host cell.6. The method of claim 5 , wherein said host cell is a lycopene-producing host cell claim 5 , preferably wherein said lycopene-producing host cell is capable of producing lycopene at levels of at least 0.5 mg/g CDW claim 5 , further preferably claim 5 , wherein the lycopene producing host cell comprises the plasmid pAC-LYC.7. The method of claim 6 , wherein said one or more proteins of (a) are capable of catalysing the conversion of lycopene to flavuxanthin.8. The method of claim 7 , wherein said one or ...

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30-05-2013 дата публикации

METHOD OF PRODUCING ISOPRENOID COMPOUNDS IN YEAST

Номер: US20130137138A1
Автор: Hansen Jorgen
Принадлежит:

Yeast strains capable of increased prenyl phosphate production are provided, enabling increased terpenoid molecule production. Heterologous yeast strains with high prenyl phosphate availability are prepared using one or both of two different strategies for increasing the availability of prenyl phosphates for terpenoid production. First, by co-expressing multiple mevalonate pathway gene analogs, a novel heterologous combination of genes results, some of which increases the inherent availability of prenyl phosphates in yeast. Second, by expressing the non-endogenous enzyme ATP citrate lysase (ACL), a buildup of high cytosolic concentration of acetyl-CoA is produced in the cytosol of 1. A method of producing isoprenoid compounds in a yeast cell , the method comprising cultivating a yeast cell in a suitable medium , the yeast cell comprisinga) one or more heterologous nucleotide sequences encoding MEV-1 (SEQ ID. NO. 36), MEV-6 (SEQ ID. NO. 41), MEV-15 (SEQ ID. NO. 50), MEV-18 (SEQ ID. NO. 53), MEV-21 (SEQ ID. NO. 56), or MEV-23 (SEQ ID. NO. 58); and/orb) reduced inherent ACO1 expression relative to an unaltered yeast cell.24-. (canceled)5. A method for making a yeast host cell with increased synthesis of isoprenoid compounds relative to an unaltered yeast cell , the method comprisinga) introducing one or more heterologous nucleotide sequences encoding MEV-1 (SEQ ID. NO. 36), MEV-6 (SEQ ID. NO. 41), MEV-15 (SEQ ID. NO. 50), MEV-18 (SEQ ID. NO. 53), MEV-21 (SEQ ID. NO. 56), or MEV-23 (SEQ ID. NO. 58) and/orb) reducing inherent ACO1 expression relative to an unaltered yeast cell.6. A yeast host cell comprising a) one or more heterologous nucleotide sequences encoding MEV-1 (SEQ ID. NO. 36) , MEV-6 (SEQ ID. NO. 41) , MEV-15 (SEQ ID. NO. 50) , MEV-18 (SEQ ID. NO. 53) , MEV-21 (SEQ ID. NO. 56) , or MEV-23 (SEQ ID. NO. 58); and/orb) reduced inherent ACO1 expression relative to an unaltered yeast cell.7. The method of claim 1 , wherein the yeast cell further comprises one or ...

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06-06-2013 дата публикации

METHOD FOR PRODUCING PEPTIDE

Номер: US20130143262A1
Принадлежит: AJINOMOTO CO., INC.

Peptides may be produced by allowing (A) a first amino acid or peptide, which is converted into its ionic liquid form, (B) a second amino acid or peptide, and (C) a peptide hydrolase to simultaneously exist in a single reaction system, wherein the first amino acid or peptide, which is converted into its ionic liquid form, is used as both a reaction solvent and a reaction starting material; and forming a peptide bond between the first amino acid or peptide and the second amino acid or peptide. By such a process, it is possible to synthesize a peptide at a high concentration and at a high yield, and the method is excellent for producing peptides on an industrial scale. 1. A method for producing a peptide , which comprises:(a) allowing (A) a first amino acid or peptide, which is converted into its ionic liquid form, (B) a second amino acid or peptide, and (C) a peptide hydrolase to simultaneously exist in a single reaction system, wherein the first amino acid or peptide, which is converted into its ionic liquid form, is used as both a reaction solvent and a reaction starting material; and(b) forming a peptide bond between said first amino acid or peptide and said second amino acid or peptide.2. A method according to claim 1 , wherein said second amino acid or peptide (B) is converted into its ionic liquid form.3. A method according to claim 2 , wherein water is present in said reaction system.4. A method according to claim 1 , wherein said first amino acid or peptide of (A) is protected at its amino group or carboxyl group.5. A method according to claim 2 , wherein said first amino acid or peptide of (A) is protected at its amino group or carboxyl group.6. A method according to claim 3 , wherein said first amino acid or peptide of (A) is protected at its amino group or carboxyl group.7. A method according to claim 1 , wherein (A) said first amino acid or peptide claim 1 , which is converted into its ionic liquid form claim 1 , is a carboxylate.8. A method according to ...

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06-06-2013 дата публикации

PRODUCTION AND PURIFICATION OF IL-29

Номер: US20130143270A1
Принадлежит: BRISTOL-MYERS SQUIBB COMPANY

The expression vectors and methods using an expression system for the large scale production of IL-29 are described. The vectors utilize the IL-29 coding sequence with specific changes in nucleotides in order to optimize codons and mRNA secondary structure for translation in . Also included are methods of producing, purifying and pegylating an IL-29 polypeptide. 1. A method of producing IL-29 polypeptide comprising:{'i': 'E. coli', 'claim-text': (i) a translational enhancer comprising SEQ ID NO:13; and', '(ii) a nucleic acid molecule encoding an IL-29 polypeptide comprising an amino acid sequence having at least 95% identity to SEQ ID NO:6;, '(a) providing an host cell comprising an expression vector comprising the following operably linked elements{'i': 'E. coli', '(b) culturing the host cell in a growth medium in a fermentation vessel;'}{'i': 'E. coli', '(c) harvesting the host cell; and'}{'i': 'E. coli', '(d) isolating the IL-29 polypeptide from the host cell.'}2E. coli. The method of claim 1 , wherein the host cell is ompT deficient.3E. coli. The method of claim 2 , wherein the host cell is fhuA deficient.4. The method of claim 1 , wherein the nucleic acid molecule encodes an IL-29 polypeptide comprising an amino acid sequence that is at least 98% identical to SEQ ID NO:6.5. The method of claim 1 , wherein the nucleic acid molecule encodes an IL-29 polypeptide comprising SEQ ID NO:6.6. The method of claim 1 , wherein the growth medium in the fermentation vessel has a pH ranging from 6.2 to 7.2.7. The method of claim 1 , further comprising feeding a carbohydrate feed solution into the fermentation vessel.8. The method of claim 7 , wherein the carbohydrate feed solution comprises a glycerol or glucose at a concentration ranging from 10 to 30 g/L growth medium.9. The method of claim 7 , wherein the carbohydrate feed solution is fed at a feed rate of 5-15 grams of glycerol or glucose per liter per hour.10. The method of claim 7 , wherein the carbohydrate feed ...

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06-06-2013 дата публикации

METHOD FOR THE PRODUCTION OF A COMPOUND OF INTEREST

Номер: US20130144034A1
Принадлежит: DSM IP ASSETS B.V.

The present invention relates to a method for the production of a compound of interest by microbial fermentation, wherein the microbial host cell used has been modified in its genome such that it results in a deficiency in the production of at least one non-ribosomal peptide synthase. The present invention further relates to a microbial host cell that has been modified in its genome such that it results in a deficiency in the production of at least one non-ribosomal peptide synthase. The invention further relates to a compound of interest. 2. The method according to claim 1 , wherein said microbial host cell has been modified in said genome on at least one position of at least one nucleic acid sequence encoding a non-ribosomal peptide synthase having at least 30% identity with a polypeptide selected from the group consisting of the polypeptide according to SEQ ID NO: 38 claim 1 , the polypeptide according to SEQ ID NO: 34 claim 1 , the polypeptide according to SEQ ID NO: 10 claim 1 , the polypeptide according to SEQ ID NO: 14 claim 1 , the polypeptide according to SEQ ID NO: 18 claim 1 , the polypeptide according to SEQ ID NO: 22 claim 1 , and the polypeptide according to SEQ ID NO: 4 and/or wherein said microbial host cell has been modified in its genome such that it results in a reduction of the amount of at least one mRNA having at least 60% identity with a mRNA selected from the group of the mRNA according to SEQ ID NO: 37 claim 1 , the mRNA according to SEQ ID NO: 33 claim 1 , the mRNA according to SEQ ID NO: 9 claim 1 , the mRNA according to SEQ ID NO: 13 claim 1 , the mRNA according to SEQ ID NO: 17 claim 1 , the mRNA according to SEQ ID NO: 21 claim 1 , and the mRNA according to SEQ ID NO: 3.3. The method according to wherein the polypeptide is one of the polypeptide according to SEQ ID NO: 38 or the polypeptide according to SEQ ID NO: 34 and/or the mRNA is one of the mRNA according to SEQ ID NO: 37 or the mRNA according to SEQ ID NO: 33.4. The method ...

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13-06-2013 дата публикации

METHOD FOR ENHANCING LUMINESCENCE INTENSITY OF CLYTIN-II

Номер: US20130149743A1
Принадлежит: JNC CORPORATION

A method for enhancing a luminescence activity of clytin-II is provided. A codon-optimized nucleic acid is used for coding the apo-clytin-II protein, and the luminescent activity of the clytin-II is remarkably enhanced when comparing with the conventional use of the wild-type clytin-II. 1. A method for preparing apo-clytin-II protein , the method comprising:(i) preparing a recombinant expression vector that allows a codon-optimized nucleic acid for coding an apo-clytin-II protein under a regulatory control of a promoter that functions in a host cell, wherein said codon optimized nucleic acid is SEQ ID NO:3;(ii) introducing the recombinant expression vector into the host cell; and(iii) culturing the host cell, to allow the apo-clytin-II protein to be expressed.2. The method for preparing apo-clytin-II protein of further comprising:(iv) purifying the expressed codon-optimized apo-clytin-II protein from a significant amount of other intracellular proteins.3. The method for preparing apo-clytin-II protein of claim 1 , wherein a purity of the codon-optimized apo-clytin-II protein is preferably 70% or more.4. The method for preparing apo-clytin-II protein of claim 1 , wherein a purity of the codon-optimized apo-clytin-II protein is preferably 85% or more.5. The method for preparing apo-clytin-II protein of claim 1 , wherein a purity of the codon-optimized apo-clytin-II protein is preferably 95% or more.6. A method for enhancing a luminescence intensity of clytin-II claim 1 , the method comprising:combining a codon-optimized nucleic acid for coding an apo-clytin-II protein with a regulatory nucleic acid for expressing the codon-optimized nucleic acid to form a nucleic acid, wherein said codon optimized nucleic acid is SEQ ID NO:3; andintroducing the resulting nucleic acid into a host cell to code and generate the apo-clytin-II protein in which an expression level of the apo-clytin-II protein is increased so that the luminescence intensity of the clytin-II is enhanced.7. ...

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20-06-2013 дата публикации

Extracellular Matrix-Derived Gels and Related Methods

Номер: US20130156862A1

Provided are methods for preparing gelled, solubilized extracellular matrix (ECM) compositions useful as cell growth scaffolds. Also provided are compositions prepared according to the methods as well as uses for the compositions. In one embodiment a device, such as a prosthesis, is provided which comprises an inorganic matrix into which the gelled, solubilized ECM is dispersed to facilitate in-growth of cells into the ECM and thus adaptation and/or attachment of the device to a patient. 196-. (canceled)97. A method of preparing an extracellular matrix-derived gel comprising: (a) solubilizing decellularized extracellular matrix (ECM) derived from cardiac tissue by digestion with an acid protease in an acidic solution to produce digested cardiac ECM; and (b) raising the pH of the digested cardiac ECM to a pH between 7.2 and 7.8 to produce a neutralized digest solution , and (c) gelling the neutralized digest solution at a temperature greater than 25° C.98. The method of claim 97 , wherein the ECM is not dialyzed or subjected to a cross-linking process prior to the gelling step (c).99. The method of claim 97 , wherein the ECM is intact ECM.100. The method of claim 97 , wherein the neutralized digest solution is maintained at or below 25° C. before the gelling step (c).101. The method of claim 97 , wherein the protease is pepsin claim 97 , trypsin or a combination thereof.102. The method of claim 97 , wherein in the step of raising the pH of the digested cardiac ECM (b) claim 97 , a base or an isotonic buffer is added to raise the pH of the digested cardiac ECM.103. The method of claim 102 , wherein the base or isotonic buffer is NaOH or phosphate buffered saline.104. The method of claim 97 , wherein the pH is raised to 7.4 in the step of raising the pH of the digested cardiac ECM (b).105. The method of claim 97 , wherein the digest solution is gelled at 30° C. or higher.106. The method of claim 97 , wherein the digest solution is gelled at 37° C.107. The method of ...

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11-07-2013 дата публикации

Endophytic Fungi from Pteromischum SP. Plant, Compounds and Methods of Use

Номер: US20130177596A1
Принадлежит:

The present disclosure relates to endophytic fungi from higher plants such as a sp. plant, and to extracts and compounds from such fungi that have desirable biological activities, such as antifungal and immunosuppressive activities. The present disclosure further relates to compositions comprising such extracts and compounds, as well as methods of making and using the compositions. 1. (canceled)2Colletotrichum. The method according to claim 11 , wherein the endophytic fungi is a sp.3. The method according to claim 11 , wherein the fungus has biological activity against a fungal plant pathogen.4Botrytis cinerea, Sclerotinia sclerotiorumRhizoctonia solani.. The method according to claim 3 , wherein the fungal plant pathogen is at least one of claim 3 , or58-. (canceled)9. The method according to claim 11 , wherein the compound has a molecular mass of 1081.7 claim 11 , 1095.7 claim 11 , 1111.7 or 1127.7.10. The method according to claim 11 , wherein the compound includes a tetrapeptide sequence Val-Ile-Ser-Ile (SEQ ID NO: 1).11. A method of suppressing an immune response claim 11 , comprising:{'i': 'Pteromischum', 'administering to a subject in need of immunosuppression a therapeutically effective amount of a composition comprising an endophytic fungus isolated from a sp. plant, wherein the isolated endophytic fungus has antifungal or immunosuppressive activity, a crude extract of the endophytic fungus or a compound obtained from the endophytic fungus, thereby suppressing the immune response.'}12. The method of claim 11 , further comprising first selecting a subject in need of immunosuppression.13Pteromischum. A method to protect a plant against a fungal pathogen comprising treating the plant with an effective amount of a composition comprising an endophytic fungus isolated from a sp. plant claim 11 , wherein the isolated endophytic fungus has antifungal or immunosuppressive activity claim 11 , a crude extract of the endophytic fungus or a compound obtained from the ...

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11-07-2013 дата публикации

C-TERMINAL MODIFICATION OF POLYPEPTIDES

Номер: US20130177940A1
Принадлежит:

The invention relates to a mutated trypsin comprising an amino acid substitution both at position K60 and D189, and at least one more amino acid substitution by histidine at position N143 or position E151. Such trypsin mutant has a preferred cleavage site comprising the amino acids Xaa-Xaa-His, wherein Xaais L, Y or F and Xaais R or K. The invention also relates to a man-made polypeptide comprising a target peptide and the above cleavage site as well as to a method of producing C-terminally modified target peptides by using this mutated trypsin. 1. A mutated trypsin comprising an amino acid substitution at position K60 and at position D189 , and an amino acid substitution by histidine at position N143 or position E151.2. The mutated trypsin of wherein K60 is substituted by E or D.3. The mutated trypsin of wherein D189 is substituted by K claim 1 , H or R.4. (canceled)5. A method of producing a C-terminally transacylated target peptide comprising the steps of:{'sub': 1', '2', '1', '2', '1, 'providing a polypeptide comprising a target peptide and a restriction site peptide comprising the cleavage site Xaa-Xaa-His, wherein Xaais L, Y or F, and Xaais R or K, wherein said restriction site peptide overlaps with the target peptide by the amino acid Xaaat the C-terminal end of said target peptide,'}{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'sub': '1', 'bringing said peptide into contact with a trypsin mutant according to under conditions allowing for endoproteolytic cleavage after Xaa, thereby forming an endoprotease target peptide peptide-acyl-intermediate,'}adding an appropriate nucleophile, and,upon nucleophilic attack and binding of said nucleophile to the C-terminus of the target peptide, releasing the mutated trypsin from the endoprotease target peptide-acyl-intermediate.6. The method of wherein said nucleophile is selected from the group consisting of a primary amine group claim 5 , an imine group claim 5 , a secondary amine group claim 5 , a thiol group and ...

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25-07-2013 дата публикации

MODIFIED TRANSKETOLASE AND USE THEREOF

Номер: US20130189731A1
Принадлежит: DSM IP ASSETS B.V.

The present invention relates to a improved process for the biotechnological production of compounds for which ribose-5-phosphate, ribulose-5-phosphate or xylulose-5-phosphate is biosynthetic precursor like riboflavin (vitamin B), FAD, FMN, pyridoxal phosphate (vitamin B), guanosine, GMP, adenosine, AMP. The invention further pertains to the generation of the organism producing those compounds. It furthermore relates to the generation of mutated transketolases that allow normal growth on glucose but reduced growth on gluconate when introduced into the production strains and to polynucleotides encoding them. 1. A modified transketolase , wherein the amino acid sequence of the modified transketolase contains at least one mutation , so that the specific activity of the modified enzyme is modulated in comparison to the corresponding non-modified wild-type enzyme.2Bacillus subtilis. A transketolase according to wherein the amino acid sequence of the modified transketolase contains a mutation at an amino acid position that corresponds to position 357 of the transketolase as shown in SEQ ID No. 2.3. A transketolase according to wherein the wild-type amino acid arginine is replaced by histidine claim 2 , alanine claim 2 , lysine claim 2 , serine claim 2 , threonine claim 2 , leucine claim 2 , valine claim 2 , isoleucine claim 2 , methionine claim 2 , glycine claim 2 , glutamine claim 2 , or asparagine.4. A transketolase according to wherein the wild-type amino acid arginine is replaced by histidine claim 2 , alanine claim 2 , lysine claim 2 , serine claim 2 , threonine claim 2 , asparagine claim 2 , or glycine.5Bacillus, Escherichia coli, Saccharomyces cerevisiae, Ashbya gossypii, Eremothecium ashbyiCorynebacterium glutamicum.. A transketolase according to which is a modification of the wild-type transketolase from the genus or6Bacillus.. A transketolase according to wherein the wild-type transketolase is from the genus7Bacillus subtilis.. A transketolase according to ...

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01-08-2013 дата публикации

Glycosylation of Molecules

Номер: US20130195835A1
Принадлежит:

Described herein are methods and genetically engineered cells useful for producing an altered N-glycosylation form of a target molecule. Also described are methods and molecules with altered N-glycosylation useful for treating a variety of disorders such as metabolic disorders. 1. A method of producing an altered N-glycosylation form of a target protein , the method comprising:{'i': Yarrowia lipolytica', 'Arxula adeninivorans, 'providing a or an cell genetically engineered to express a protein, or a biologically active variant thereof, capable of effecting mannosyl phosphorylation of N-glycans; and'}introducing into the cell a nucleic acid encoding a target protein, wherein the cell produces the target protein in an altered N-glycosylation form.2. The method of claim 1 , further comprising isolating the altered N-glycosylation form of the target protein.3. The method of claim 1 , wherein the target protein is an exogenous protein.4. The method of claim 1 , wherein the target protein is an endogenous protein.5. The method of claim 1 , wherein the target protein is a mammalian protein.6. The method of claim 1 , wherein the target protein is a pathogen protein claim 1 , a lysosomal protein claim 1 , a growth factor claim 1 , a cytokine claim 1 , a chemokine claim 1 , or a fusion protein.7. The method of claim 1 , wherein the target protein is a protein associated with a lysosomal storage disorder (LSD).8. The method of claim 7 , wherein the lysosomal storage disorder is Gaucher disease claim 7 , Tay-Sachs disease claim 7 , Pompe disease claim 7 , Niemann-Pick disease claim 7 , or Fabry disease.9. The method of claims 7 , wherein the target protein is glucocerebrosidase claims 7 , alpha galactosidase claims 7 , or galactocerebrosidase.10. The method of claim 7 , wherein the target protein is selected from the group consisting of alpha-L-iduronidase claim 7 , beta-D-galactosidase claim 7 , beta-glucosidase claim 7 , beta-hexosaminidase claim 7 , beta-D-mannosidase claim ...

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01-08-2013 дата публикации

Device and method for cell free analytical and preparative protein synthesis

Номер: US20130196372A1
Принадлежит: Ionovation GmbH

A device is disclosed which contains one or more pores 10. The pores 10 in turn contain one or more translocase proteins which from a translocation system 20. At the pores 10, two zones, the cis zone 50 and the trans zone 60, are separated from one another on a support body 90 by means of translocation systems 20 in such a way that only those proteins that are recognized by the translocation systems 20 due to specific molecular signals can exclusively pass over from the cis zone 50 into the trans zone 60.

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15-08-2013 дата публикации

WHOLE EGG PROTEIN PEPTIDES, PREPARATION METHOD AND USE THEREOF

Номер: US20130209497A1
Автор: Zou Yuandong

Provided are whole egg protein peptides and the preparation method thereof, wherein the whole egg protein peptides are obtained by adopting compound proteases composed of pawpaw protease, fig protease and pineapple protease to enzymatically hydrolyze the whole egg protein powder. The whole egg protein peptides can be used for manufacture of products for enhancing immunity. 1. A method of preparing whole egg protein peptides , comprising:enzymatically degrading whole egg protein powders using a compound plant protease, so as to obtain the whole egg protein peptides;wherein the compound plant protease consists of pawpaw protease, fig protease and pineapple protease.2. The method according to wherein the compound plant proteases have an enzyme activity ratio of pawpaw protease claim 1 , fig protease and pineapple protease of (1-1.2 million)U:(0.3-0.4 million)U:(5.6-7 million)U.3. The method according to wherein the whole egg protein powders are prepared from at least one of following bird eggs as a raw material: chicken claim 1 , duck claim 1 , goose claim 1 , quail claim 1 , sparrow claim 1 , pigeon claim 1 , turtledove and ostrich eggs.4. The method according to wherein claim 1 , in said enzymatically degrading whole egg protein powders claim 1 , the whole egg protein powders are present with water claim 1 , and a ratio of whole egg protein powders to water by mass part is 1:8-10; amounts of enzymes required for enzymatically degrading per gram of the whole egg protein powders are: 50-60 thousand U of pawpaw protease claim 1 , 15-20 thousand U of fig protease claim 1 , and 280-350 thousand U of pineapple protease; and the enzymatic degradation is performed at a temperature of 48-50° C. claim 1 , for a period of 3-4 hours claim 1 , with a pH value of 7.5-8.5.5. The method according to further comprising:sterilizing a hydrolysate obtained from said enzymatic degradation of the whole egg protein powders, and inactivating enzymes in the hydrolysate; andcooling and ...

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22-08-2013 дата публикации

Novel artificial translation/synthesis system

Номер: US20130217599A1
Принадлежит: University of Tokyo NUC

A new artificial translation-synthesis system of adding tRNAs binding special amino acids to the in vitro translation system and synthesizing peptides with special amino acids incorporated thereto according to a dual genetic code table and an artificial codon box division.

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22-08-2013 дата публикации

Method to Improve Glycosylation Profile and to Induce Maximal Cytotoxicity for Antibody

Номер: US20130217863A1

The present invention relates to the production of recombinant glycoproteins or antibodies that have an improved glycosylation profile and effector functions such as ADCC and/or CDC. The present invention is in particular related to the production of glycoproteins or antibodies having a valuable glycosylation profile, especially a low fucose level and/or a high oligomannose level and/or presence of sialic acid to the glycans. 1. A method to produce in wild type rodent cells , more preferably wild type CHO cells , an antibody having ADCC and CDC function and containing an Fc region having a low fucose level and/or a high oligomannose level and/or high level of sialylated glycoforms , comprising engineering or using a nucleic acid sequence coding for a variant Fc region wherein this variant region comprises amino acid substitutions at the amino acid positions 243 , 292 , 300 , 305 , 326 , and 396 or at the positions 243 , 292 , 300 , 305 , 326 , 333 and 396 of the human IgG Fc region.2. The method of claim 1 , wherein the proportion of non-fucosylated Fc or antibodies represent at least 20% of the Fc or antibodies and/or the proportion Fc or antibodies featured by a higher level of oligomannoses represent at least 20% claim 1 , of the Fc or antibodies and/or the proportion of Fc or antibodies featured by a higher level of sialylated glycoforms represent at least 1.5%.3. A method for the production of an antibody having ADCC and CDC functions claim 1 , and a glycosylation profile characterized by a low fucose level and a high oligomannose level claim 1 , the method comprising the production of an antibody having one or two claim 1 , preferably two claim 1 , human IgG Fc region having amino acid substitutions at positions 243 claim 1 , 292 claim 1 , 300 claim 1 , 305 claim 1 , 326 and 396 or at positions 243 claim 1 , 292 claim 1 , 300 claim 1 , 305 claim 1 , 326 claim 1 , 333 and 396 and wherein the antibody is produced in a wild type rodent cell.4. The method of claim ...

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29-08-2013 дата публикации

METHOD OF PRODUCING FR901228

Номер: US20130225507A1
Принадлежит: ATSELLAS PHARMA INC.

Depsipeptides and congeners thereof are disclosed having structure (I), wherein m, n, p, q, X, R1, R2 and R3 are as defined herein. These compounds, including FR901228, have activity as, for example, immunosuppressants, as well as for the prevention or treatment of patients suffering or at risk of suffering from inflammatory, autoimmune or immune system-related diseases including graft-versus-host disease and enhancement of graft/tissue survival following transplant. Also provided are methods for inhibiting lymphocyte activation, proliferation, and/or suppression of IL-2 secretion. 2. The pharmaceutical composition of claim 1 , wherein the type B crystalline form of FR901228 exhibits a powder X-ray diffraction pattern substantially similar to .3. The pharmaceutical composition of claim 1 , wherein the type B crystalline form of FR901228 exhibits one or more of the following peaks in a powder X-ray diffraction pattern: about 7.1 claim 1 , about 11.7 claim 1 , about 11.9 claim 1 , and about 15.1 degrees 2-theta. This application is a continuation of U.S. patent application Ser. No. 13/473,635, filed May 17, 2012, which, in turn, is a continuation of U.S. patent application Ser. No. 12/495,239, filed Jun. 30, 2009, which, in turn, is a continuation of U.S. patent application Ser. No. 12/049,746, filed Mar. 17, 2008, which, in turn, is a continuation of U.S. patent application Ser. No. 10/362,359, which is the U.S. National Stage of International Application No. PCT/JP01/07191, filed Aug. 22, 2001, the disclosures of which are incorporated herein by reference in their entireties. This application claims priority to Japanese Patent Application No. JP 2000-265414, filed Sep. 1, 2000, the disclosure of which is incorporated herein by reference in its entirety.The present invention relates to a method of producing FR901228 which is useful as an antibacterial agent and an antitumor agent. More particularly, it relates to a method of producing FR901228 which comprises ...

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05-09-2013 дата публикации

TRUNCATED L1 PROTEIN OF HUMAN PAPILLOMAVIRUS TYPE 52

Номер: US20130230548A1
Принадлежит: XIAMEN UNIVERSITY

Provided is a truncated L1 protein of Human Papillomavirus (HPV) Type 52 which, compared to a wild type HPV52 L1 protein, is truncated by 27-42 amino acids at the N-terminal. Also provided are a coding sequence of the truncated HPV52 L1 protein, a virus-like particle (VLP) comprising the protein, and a method of preparing the protein and the VLP using an expression system. The truncated HPV52 L1 protein and an assembled VLP can be used to prevent an HPV52 infection and a disease caused by HPV52 infection, such as cervical cancer. 113.-. (canceled)14. A truncated HPV52 L1 protein or variants thereof , wherein the protein is 27-42 amino acids in length , truncated at its N-terminal , when compared with wild type HPV52 L1 protein.15. The truncated HPV52 L1 protein as claimed in claim 14 , wherein the protein has a number of amino acids selected from the group consisting of claim 14 , 27 claim 14 , 28 claim 14 , 29 claim 14 , 30 claim 14 , 31 claim 14 , 32 claim 14 , 33 claim 14 , 34 claim 14 , 35 claim 14 , 36 claim 14 , 37 claim 14 , 38 claim 14 , 39 claim 14 , 40 claim 14 , 41 claim 14 , and 42 amino acids claim 14 , truncated at its N-terminal claim 14 , when compared with wild type HPV52 L1 protein.16. The truncated HPV52 L1 protein as claimed in claim 15 , wherein the protein has a number of amino acids selected from the group consisting claim 15 , 27 claim 15 , 35 claim 15 , 38 claim 15 , 40 claim 15 , and 42 amino acids claim 15 , truncated at its N-terminal claim 15 , as compared with wild type HPV52 L1 protein.17. The truncated HPV52 L1 protein as claimed in claim 14 , wherein the protein has an amino acid sequence selected from the group consisting of SEQ ID NO: 1 claim 14 , SEQ ID NO: 2 claim 14 , SEQ ID NO: 3 claim 14 , SEQ ID NO: 4 claim 14 , SEQ ID NO: 5 claim 14 , SEQ ID NO: 6 claim 14 , SEQ ID NO: 7 claim 14 , SEQ ID NO: 8 claim 14 , SEQ ID NO: 9 claim 14 , SEQ ID NO: 10 claim 14 , SEQ ID NO: 11 claim 14 , SEQ ID NO: 12 claim 14 , and SEQ ID NO: 13.18. ...

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12-09-2013 дата публикации

ANIONIC EXCHANGE-HYDROPHOBIC MIXED MODE

Номер: US20130237692A1
Автор: Frost Russell, Liao Jiali
Принадлежит: Bio-Rad Laboratories, Inc.

Anion exchange-hydrophobic mixed mode ligands and methods of their use are provided. 2. The chromatographic solid support of claim 1 , wherein Rand Rare hydrogen.3. The chromatographic solid support of claim 1 , wherein Ris an alkyl comprising 1-4 carbons.4. The chromatographic solid support of claim 1 , wherein Ris an alkyl comprising 2 carbons.5. The chromatographic solid support of claim 1 , wherein the aryl is a phenyl.6. The chromatographic solid support of claim 1 , wherein the aryl is a substituted phenyl.7. The chromatographic solid support of claim 6 , wherein the substituted phenyl is an alkyl-substituted phenyl.8. The chromatographic solid support of claim 1 , wherein Rand/or Rare an alkyl comprising 1-6 carbons and Ar is an alkyl-substituted phenyl.9. The chromatographic solid support of claim 1 , wherein the ligand is N-phenylethylenediamine or a salt thereof.10. The chromatographic solid support of claim 1 , wherein the ligand is 2-phenoxyethylamine or a salt thereof.11. The chromatographic solid support of claim 1 , wherein X is absent.12. The chromatographic solid support of claim 1 , wherein X is a spacer.13. A method of purifying a biomolecule from a sample claim 1 , comprising contacting the sample to the chromatographic solid support linked to the ligand of ; andcollecting a purified biomolecule.14. The method of claim 13 , wherein the biomolecule is a protein.15. The biomolecules of claim 14 , wherein the protein is an antibody.16. The method of claim 13 , wherein the sample comprises monomeric antibodies and aggregates claim 13 , and monomeric antibodies are separated from aggregates claim 13 , thereby resulting in purification of monomeric antibodies.17. The method of claim 13 , wherein the purified biomolecule is a monomeric antibody.18. The method of claim 17 , wherein monomeric antibodies of the sample are immobilized on the chromatographic solid support and subsequently eluted.19. The method of claim 17 , wherein monomeric antibodies of ...

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26-09-2013 дата публикации

HYDROPHOBIZED PROTEIN HYDROLYSATE

Номер: US20130251658A1
Принадлежит: EVONIK GOLDSCHMIDT GMBH

The invention relates to enzymatically hydrophobicised protein hydrolysates, their production and use, and to cosmetic preparations comprising these. 2. The alkylated peptide mixture according to claim 1 , wherein said at least one protein is selected from the group consisting of isolated plant storage proteins claim 1 , animal proteins claim 1 , and microbial proteins.3. The alkylated peptide mixture according to claim 1 , wherein the hydrolysis in process step A) is catalysed by at least one acid or at least one enzyme.4. The alkylated peptide mixture according to claim 1 , wherein said protein hydrolysate in process step A) has an average molecular weight of 203 g/mol to 100 claim 1 ,000 g/mol.5. The alkylated peptide mixture according claim 1 , wherein said protein hydrolysate in process step A) is selected from Meripro 810 claim 1 , Meripro 711 claim 1 , Naturalys W claim 1 , Cropeptide W claim 1 , Hydrotriticum 2000 claim 1 , Tritisol claim 1 , Tritisol XM claim 1 , Hydrosoy 2000 claim 1 , Gluadin W20 claim 1 , Gluadin WLM claim 1 , AMCO HCA411 claim 1 , and HLA-198.6Bacillus subtilisStreptoverticillium mombaraensis.. The alkylated peptide mixture according to claim 1 , wherein said transglutaminase in process step B) is isolated from or7. The alkylated peptide mixture according to claim 1 , wherein said radicals Rand Rare independently selected from the group consisting of alkyl and alkenyl radicals.8. The alkylated peptide mixture according to claim 1 , wherein claim 1 , in process step A) claim 1 , the hydrolysis is catalysed by at least one enzyme claim 1 , and process step A) and process step B) are conducted simultaneously.10. A cosmetic claim 1 , dermatological claim 1 , pharmaceutical crop protection claim 1 , care claim 1 , cleaning claim 1 , or surfactant formulation comprising at least one alkylated peptide mixture according to .11. An emulsifier claim 1 , dispersion auxiliary claim 1 , conditioner for skin and hair claim 1 , foam former claim 1 , ...

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26-09-2013 дата публикации

METHOD FOR PREPARING ACTIVE PEPTIDES FROM CORN GERM PROTEINS

Номер: US20130252877A1

The present invention discloses a method for producing antihypertensive active peptides with corn germ protein as the material. The method comprises an alkali-heat treatment and continuous enzymolysis of the corn germ protein. The components with molecular weight less than 1000 Da in the active peptides obtained according to the present method account for more than 92%, and alanine-tyrosine (Ala-Tyr, AY) as the characteristic peptide fragments in the antihypertensive peptides accounts for more than 0.6%, so that the active peptides have a good ACE inhibitory activity in vitro as well as stability against temperature, pH and major gastrointestinal digestive enzymes, and have a significant effect of lowering blood pressure on spontaneous hypertension rats in vivo. The active peptides can be applied as a new functional nutrient to development and production of food, health food and pharmaceutical. 1. A method for preparing active peptides from corn germ protein powders , including the following steps:adding the corn germ protein powders to a reaction tank, mixing the corn germ protein powders with water to form a first feed liquid, and the feed liquid being adjusted to be alkalescent, heated to 50˜90° C., and stirred at this temperature for 20˜60 min;centrifugalizing the alkalescent first feed liquid in the reaction tank and collecting slag from the alkalescent first feed liquid;mixing the slag with water to form a second feed liquid, the second feed liquid being heated to 50˜90° C. and centrifugalized, and collecting slag from the second feed liquid; the same processing is repeated at least twice to obtain purified slag;the purified slag being mixed with water at a water-slag ratio of 100:40˜60, stirred, and subjected to a first enzymolysis and a second enzymolysis by alkali protease and compound protease in sequence to obtain an enzymatic hydrolysate, wherein the compound protease is comprised of papain and neutral protease; andheating the enzymatic hydrolysate to ...

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10-10-2013 дата публикации

PLANT PROTEIN HYDROLYSATES

Номер: US20130267683A1
Принадлежит: NESTEC S.A.

A membrane reactor for the manufacture of plant protein hydrolysates, the membrane reactor comprising a substrate vessel adapted to provide a plant protein substrate to an enzyme source, a continuously stirred reactor comprising the enzyme source, and an ultrafiltration module comprising a membrane with a molecular cut-off wherein the membrane is adapted to allow passage of the plant protein hydrolysate while retaining the enzyme. 1. A membrane reactor for the manufacture of plant protein hydrolysates , the membrane reactor comprising:a substrate vessel adapted to provide a plant protein substrate to an enzyme source;a continuously stirred reactor comprising the enzyme source; andan ultrafiltration module comprising a membrane with a molecular cut-off wherein the membrane is adapted to allow passage of the plant protein hydrolysate while retaining the enzyme.2. The membrane reactor as claimed in claim 1 , further comprising:a first circulation loop enabling a mixture of the plant protein substrate and enzyme source to be transferred from the continuously stirred reactor to the ultrafiltration module and at least some of the mixture to be returned to the continuously stirred reactor; anda second circulation loop enabling the mixture received from the first circulation loop to be circulated through or over the membrane and at least some of the mixture to be returned to the first circulation loop.3. The membrane reactor as claimed in claim 2 , wherein the first circulation loop operates at or close to atmospheric pressure and the second circulation loop operates at a pressure of 1 to 8 bar.4. The membrane reactor as claimed in claim 1 , comprising a heating device adapted to maintain a temperature of the content of the continuously stirred reactor between 25° C. and 75° C.5. The membrane reactor as claimed in claim 1 , comprising an electro dialysis system.6. The membrane reactor as claimed in claim 1 , comprising a separation device claim 1 , capable of separating ...

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17-10-2013 дата публикации

PROTEIN CONCENTRATE FROM STARCH CONTAINING GRAINS: COMPOSITION, METHOD OF MAKING, AND USES THEREOF

Номер: US20130274443A1
Принадлежит:

The present invention relates to methods of producing a protein concentrate from a starch containing grain and uses thereof. In an exemplary embodiment, the protein concentrate produced is used to prepare an aquaculture feed. 1. A process for producing a protein concentrate from a starch containing grain or oil seed , the method comprising:(i) grinding the starch containing grain to produce a ground starch containing grain;(ii) slurrying the ground starch containing grain with water to prepare a slurry comprising starch and glucans;(iii) solublizing starch and glucans comprising the slurry with enzymes to provide a solublized slurry;(iv) adding a fermentation organism to the solublized slurry;(v) fermenting the solublized slurry comprising the fermentation organism until fermentation is complete, thereby producing a fermented slurry;(vi) separating the fermented slurry into solid and liquid fractions;(vii) recovering the solid and liquid fractions;(viii) drying the recovered solid fraction at a temperature below that which would denature or damage proteins;thereby producing a protein concentrate.2. The process of claim 1 , wherein solublizing starch and glucans with enzymes produces glucose.3. The process of claim 1 , wherein the process is a no-cook process.4. The process of claim 1 , wherein the process is a cooking process.5. The method of claim 1 , further comprising:(ix) distilling the recovered liquid fraction to recover a fermentation product.6. The method of claim 5 , wherein the fermentation product is ethanol.7. The method of claim 1 , wherein the starch containing grain is a member selected from the group consisting of barley claim 1 , wheat claim 1 , oats claim 1 , corn claim 1 , rye claim 1 , tritcale claim 1 , sorghum claim 1 , soybeans claim 1 , and soymeal claim 1 , flax claim 1 , camelina claim 1 , or a combination thereof.8. The method of claim 7 , wherein the starch containing grain is barley.9. The method of claim 8 , wherein the barley is from a ...

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24-10-2013 дата публикации

METHODS OF ACTIVATING CLOSTRIDIAL TOXINS

Номер: US20130281666A1
Принадлежит:

The specification discloses modified Clostridial toxins comprising an exogenous Clostridial toxin di-chain loop protease cleavage site located within the di-chain loop region; polynucleotide molecules encoding such modified Clostridial toxins; method of producing such modified Clostridial toxins, method of activating such modified Clostridial toxins and methods of activating recombinantly-expressed Clostridial toxins. 1. A modified Clostridial toxin comprising an exogenous BoNT/A di-chain loop region including a BoNT/A di-chain protease cleavage site;wherein the Clostridial toxin is a BoNT/B; and,wherein the BoNT/A di-chain loop region replaces an endogenous Clostridial toxin di-chain loop region.2. A polynucleotide molecule encoding a modified Clostridial toxin according to .3. A method of producing a modified Clostridial toxin comprising the step of expressing in a cell a polynucleotide molecule according to claim 2 , wherein expression from the polynucleotide molecule produces the encoded modified Clostridial toxin.4. A method of producing a modified Clostridial toxin comprising the steps of:{'claim-ref': {'@idref': 'CLM-00002', 'claim 2'}, 'a. oducing into a cell a polynucleotide molecule as defined in ; and'}b. expressing the polynucleotide molecule, wherein expression from the polynucleotide molecule produces the encoded modified Clostridial toxin. This application is a continuation of and claims priority pursuant to 35 U.S.C. §120 to U.S. patent application Ser. No. 12/669,447, filed Jan. 15, 2010, which claims priority pursuant to 35 U.S.C. 371 to application PCT/US08/68504, filed Jun. 27, 2008, which claims priority pursuant to 35 U.S.C. §119(e) to U.S. Provisional Patent Application Ser. No. 60/952,112 filed Jul. 26, 2007, all incorporated entirely by reference.The ability of Clostridial toxins, such as, e.g., Botulinum neurotoxins (BoNTs), BoNT/A, BoNT/B, BoNT/C1, BoNT/D, BoNT/E, BoNT/F and BoNT/G, Tetanus neurotoxin (TeNT), Baratium neurotoxin (BaNT) and ...

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14-11-2013 дата публикации

COMPOSITIONS AND METHODS OF PRODUCING ENTEROKINASE IN YEAST

Номер: US20130302853A1
Принадлежит:

The present specification disclose polynucleotide molecules encoding an enterokinase, yeast expression constructs including a yeast expression vector and a polynucleotide molecules encoding an enterokinase, yeast cells comprising such a yeast expression construct, methods of producing enterokinase using such yeast cells, and method of cleaving or preparing a recombinant polypeptide using an enterokinase produced by such methods. 1. A method of cleaving a recombinant polypeptide , the method comprising the step contacting a polypeptide comprising a cleavage site of SEQ ID NO:1 with an enterokinase , wherein the enterokinase is produced by expressing in a yeast cell comprising a yeast expression construct comprising SEQ ID NO:4 , wherein contacting the recombinant polypeptide with the enterokinase results in specific cleavage of SEQ ID NO:1.2. A method of cleaving a recombinant polypeptide , the method comprising the step contacting a polypeptide comprising a cleavage site of SEQ ID NO:1 with an enterokinase , wherein the enterokinase is produced by expressing in a yeast cell comprising a yeast expression construct comprising SEQ ID NO:6 , wherein contacting the recombinant polypeptide with the enterokinase results in specific cleavage of SEQ ID NO:1. This application is a Continuation of U.S. patent application Ser. No. 13/303,691, filed Nov. 23, 2011, now U.S. Pat. No. 8,497,111, which claims priority pursuant to 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 61/416,622, filed on Nov. 23, 2010, both incorporated by reference.Enterokinase (EK, also known as Enteropeptidase (EP); EC 3.4.21.9) is a heterodimeric glycoprotein produced by cells of the duodenum. Part of the chymotrypsin-clan of serine proteases, it is secreted from intestinal glands (the crypts of Lieberkühn) following the entry of ingested food passing from the stomach and present in the duodenal and jejunal mucosa. Involved in the digestion of dietary proteins, EK catalyzes the cleavage of ...

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14-11-2013 дата публикации

METHOD FOR THE PREPARATION OF SURFACTANT PEPTIDES

Номер: US20130303726A1
Принадлежит:

Surfactant-protein C peptides (SP-C peptides) may be prepared by the heterologous expression of a fusion protein of an SP-C peptide and a maltose binding protein. 1. An expression cassette , comprising:(a) a polynucleotide sequence encoding an SP-C peptide;(b) a polynucleotide sequence encoding an MBP protein; and(c) a polynucleotide sequence encoding a linker peptide,wherein said a polynucleotide sequence encoding a linker peptide encodes a protease cleavage site and is located between said polynucleotide sequence encoding an SP-C peptide and said polynucleotide sequence encoding an MBP protein, andsaid encoding polynucleotide sequence being operatively linked to a promoter sequence suitable for the expression in a prokaryotic cell.2. An expression cassette according to claim 1 , wherein said SP-C peptide is SP-C33(Leu) and has the sequence shown in SEQ ID NO:1.3. An expression cassette according to claim 1 , wherein the MBP protein has the sequence shown in SEQ ID NO:2.4. An expression cassette according to claim 1 , wherein said polynucleotide sequence encoding an SPC peptide has the sequence shown in SEQ ID NO:3.5. An expression cassette according to claim 1 , wherein said polynucleotide sequence encoding an MBP protein has the sequence shown in SEQ ID NO:4.6. An expression cassette according to claim 1 , wherein said linker peptide has the amino acid sequence shown in SEQ ID NO:5.7. An expression cassette according to claim 1 , wherein said polynucleotide sequence encoding a linker peptide has the sequence shown in SEQ ID NO:5.8. An expression cassette according to claim 1 , wherein said polynucleotide sequence encoding an MBP protein is located at the N-terminus.9. An expression cassette according to claim 1 , wherein said polynucleotide sequence encoding an SP-C peptide is located at the C-terminus.10. An expression cassette according to claim 1 , wherein said polynucleotide sequence encoding an MBP protein is located at the N-terminus and said polynucleotide ...

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21-11-2013 дата публикации

HETEROTROPHIC MICROBIAL PRODUCTION OF XANTHOPHYLL PIGMENTS

Номер: US20130309719A1
Автор: Griffiths Hywel David
Принадлежит: Photonz Corporation Limited

The invention relates to a microbial biomass wherein the biomass comprises at least one xanthophyll selected from any one or more of fucoxanthin, diatoxanthin and diadinoxanthin. The invention further relates to a process for producing such a microbial biomass. Further the invention relates to compositions comprising at least one xanthophyll selected from any one or more of fucoxanthin, diatoxanthin and diadinoxanthin and processes for producing such compositions. 1. A microbial biomass , produced from a heterotrophic fermentation , the biomass comprising at least one xanthophyll selected from any one or more of fucoxanthin , diatoxanthin and diadinoxanthin.2. The microbial biomass of wherein the xanthophyll is fucoxanthin.35-. (canceled)6. The microbial biomass of wherein the xanthophyll is present at levels equal to or greater than about 0.1% of dry cell weight of the biomass.79-. (canceled)10. The microbial biomass of wherein the microbial biomass is a marine diatom biomass.11. (canceled)12. A process for producing a microbial biomass claim 1 , wherein the microbial biomass comprises at least one xanthophyll selected from any one or more of fucoxanthin claim 1 , diatoxanthin and diadinoxanthin claim 1 , the process comprising the steps of:cultivating a microorganism in heterotrophic culture to produce the biomass; andrecovering said biomass.13. The process of wherein the xanthophyll is fucoxanthin.1415-. (canceled)16. The process of wherein the xanthophyll is present at levels equal to or greater than about 0.1% of dry cell weight of the biomass.1719-. (canceled)20. The process of wherein the microbial biomass is a marine diatom biomass.21. (canceled)22. The process of wherein at least about 1 mg of the xanthophyll is produced per litre of culture per hour.23. (canceled)24. The process of wherein the step of cultivating the microorganism in heterotrophic culture comprises a culture phase in which cells are grown under conditions in which organic carbon is used as ...

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21-11-2013 дата публикации

USE OF BROWNED GLUCOSE AS A FEED SUBSTRATE

Номер: US20130309722A1
Автор: Banke Niels
Принадлежит: NOVOZYMES A/S

A method for fermenting a microorganism, producing a compound of interest, in a culture medium comprising: adding a browned glucose solution to the culture medium, wherein the browned glucose solution is a glucose solution that has been acid treated and heated to a temperature of at least 90 degrees Celsius, and wherein the glucose solution has a concentration of at least 500 g/l. 1. A method for fermenting a microorganism , producing a compound of interest , in a culture medium comprising:adding a browned glucose solution to the culture medium, wherein the browned glucose solution is a glucose solution that has been acid treated and heated to a temperature of at least 90 degrees Celsius, and wherein the glucose solution has a concentration of at least 500 g/l.2. The method according to claim 1 , wherein the microorganism is a bacterium or a fungus.3Bacillus, Streptomyces, EscherichiaPseudomonas.. The method according to claim 2 , wherein the bacterium is selected from the group consisting of claim 2 , and4Achlya, Aspergillus, Cephalosporium, Cochliobolus, Endothia, Fusarium, Humicola, Mucor, Neurospora, Penicillium, Podospora, PyriculariaTrichoderma.. The method according to claim 2 , wherein the fungus is selected from the group consisting of claim 2 , and5. The method according to claim 1 , wherein the compound of interest is a secondary metabolite.6. The method according to claim 1 , wherein the compound of interest is a protein.7. The method according to claim 6 , wherein the protein is an enzyme.8. The method according to claim 1 , wherein the fermentation is a batch claim 1 , a fed batch claim 1 , a repeated fed batch or a continuous fermentation.9. The method according to claim 1 , wherein the acid treatment results in a glucose solution with a pH less than pH 4.5.10. The method according to claim 1 , wherein the glucose solution is acid treated and thereafter heated to a temperature of at least 90 degrees Celsius.11. The method according to claim 1 , ...

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21-11-2013 дата публикации

Method and apparatus for producing cells and fat soluble materials by cell culture

Номер: US20130309757A1
Автор: Sung-Chun Kim
Принадлежит: Individual

The present invention relates to a method and apparatus for producing cells without injury and fat-soluble materials by from cell culturing in an inexpensive and highly efficient manner. The apparatus according to the present invention comprises a culturing device 10 , a solvent device 20 , a mixing device 30 , a separation device 40 , a fractionation device 50 , a cell accommodation device 60 , and a fat-soluble material solvent accommodation device 70.

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05-12-2013 дата публикации

INTEGRATED CARBON CAPTURE AND ALGAE CULTURE

Номер: US20130319059A1
Принадлежит: WASHINGTON STATE UNIVERSITY

The feasibility of using COfrom a concentrated source to grow microalgae is limited by the high cost of COcapture and transportation, as well as significant COloss during algae culture. Another challenge is the inability of algae in using COduring night while COis continuously produced from the source. To address these challenges, this invention provides a process in which COis captured as bicarbonate and used as feedstock for algae culture. Then the carbonate is regenerated in the algae culture process as absorbent to capture more CO, which is converted to bicarbonate for use as feedstock, etc. This process significantly reduces carbon capture costs since it avoids the energy for carbonate regeneration. Also, transporting a solid or aqueous bicarbonate solution has a much lower cost than transporting compressed CO, and using bicarbonate provides a better alternative for COdelivery to algae culture systems than supplying COgas. 1. An integrated method culturing algae or cyanobacteria , comprising the steps of{'sub': 2', '2, 'i) capturing COfrom a source of CO;'}{'sub': '2', 'ii) converting captured COinto bicarbonate;'}iii) culturing alkaliphilic algae or alkaliphilic cyanobacteria using said bicarbonate as a carbon source to produce algal bioproducts;{'sub': '2', 'iv) using spent medium from said step of culturing as said source of COin said step of capturing; and'}v) repeating steps i) to iv).2. The method of claim 1 , wherein said bicarbonate is in a form selected from the group consisting of solid bicarbonate and a liquid bicarbonate solution.3SynechocystisCyanotheceMicrocoleusEuhalotheceSpirulina. The method of claim 1 , wherein said alkaliphilic cyanobacteria are selected from the group consisting of sp. claim 1 , sp. claim 1 , sp. claim 1 , sp. and sp.4ChlorellaDunaliella.. The method of claim 1 , wherein said alkaliphilic algae are eukaryotic microalgae selected from the group consisting of and5. The method of claim 1 , wherein culture medium used in said ...

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12-12-2013 дата публикации

Cell Lysis Process

Номер: US20130330767A1

A process for cell lysis is provided. The process comprises passing a mixture comprising a suspension of cells and a lysis reagent through a flow-through reactor, wherein the mixture passes through the reactor with pulsed or superimposed oscillating flow. The process is preferably employed for the preparation of biological products, such as pDNA or inclusion bodies. 1. A process for cell lysis which comprises passing a mixture comprising a suspension of cells and a lysis reagent through a flow-through reactor , wherein the mixture passes through the reactor with pulsed or superimposed oscillating flow.2. A process according to claim 1 , wherein the mixture passes through the reactor with superimposed oscillating flow.3. A process according to claim 2 , wherein the oscillating flow is at a frequency of up to 20 Hz.4. A process according to which is operated with fluid strain rates of less than 1×10s.5. A process according to claim 1 , wherein two or more flow-through reactors are employed in series.6. A process according to claim 5 , wherein a quenching reagent is added to the mixture after it has passed through the first flow-through reactor.7. A process for producing a biological product claim 5 , comprising the steps of:a) culturing host cells producing the biological product;{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'b) lysing the cells by a process according to ; and'}c) recovering the biological product.8. A process according to claim 7 , wherein the biological product is pDNA or an inclusion body.9E. coli.. A process according to either of claim 6 , wherein the host cell is10. A process according to or claim 6 , wherein the reactor is configured and operated to achieve an oscillatory Reynolds number of at least 10 claim 6 , and no more than 500.11. A process according to or claim 6 , wherein the reactor is configured and operated to achieve a net flow Reynolds number of from 5 to 50.12. A process according to or claim 6 , wherein the reactor is ...

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26-12-2013 дата публикации

EXTRACTION OF SCENEDESMUS CELL COMPONENTS

Номер: US20130344576A1
Принадлежит: DIREVO INDUSTRIAL BIOTECHNOLOGY GmbH

The technology disclosed herein relates to novel methods and compositions for the production processes of algae cell components from algae of the genus . The extractability of algae cellular components is improved and the process is optimized and accelerated by using the enzyme compositions according to the present invention. 1Scenedesmus. A method for improving the extractability of an algae cellular component from algae belonging to the genus comprising:a) subjecting the algae to an enzyme composition comprising a 1,3(4)-beta glucanase, andb) isolating the cellular composition from the algae.2. The method according to claim 1 , whereby the 1 claim 1 ,3(4)-beta glucanase is an endo-1 claim 1 ,3(4)-beta glucanase.3. The method according to claim 1 , whereby the cellular component is selected from the group consisting of pigments claim 1 , carotenoids claim 1 , starch claim 1 , lipids claim 1 , poly unsaturated fatty acids (PUFA) claim 1 , proteins claim 1 , vitamins claim 1 , and mineral nutrients.4. The method according to claim 1 , whereby the cellular component is a pigment.5. The method according to claim 1 , whereby the enzyme composition further comprises a mannanase claim 1 , preferably a 1 claim 1 ,4-beta-mannanase.6. The method according to claim 5 , whereby the 1 claim 5 ,4-beta-mannanase is an endo-1 claim 5 ,4-beta-mannanase.7. The method according to claim 1 , whereby the enzyme composition further comprises a protease.8. The method according to claim 7 , whereby the enzyme composition comprises an endo-1 claim 7 ,3(4)-beta glucanase claim 7 , an endo-1 claim 7 ,4-beta-mannanase and the protease.9. The method according to claim 1 , whereby the enzyme composition further comprises cellulases and/or a pectinases and/or a laminarinase.10Scenedesmus. A method for producing an algae cell component by using algae belonging to the genus comprising the steps:a) culturing the algae in a liquid medium,b) harvesting the algae,c) disrupting the algae cells by ...

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16-01-2014 дата публикации

METHODS FOR ISOLATING BACTERIA

Номер: US20140017724A1
Принадлежит:

The present invention relates to compositions and methods to identify novel bacteria and metabolites derived therefrom. More specifically, the invention describes a novel method to isolate bacteria producing metabolites of interest from environmental samples. Particularly, the invention discloses a method to select rare antibiotic producing bacteria. The invention can be used from any sample and allows the isolation of bacteria having e.g., pharmaceutical or agrochemical interest. 1Deinococcus. A method for producing a drug , comprising (i) culturing , in a suitable medium , a bacterium which naturally produces said drug or an intermediate thereof , (ii) collecting or purifying the drug or intermediate from the culture , and (iii) when an intermediate is produced in step (i) , converting said intermediate into said drug , wherein said bacterium is a or a related bacterium.2Deinococcus. An improved method for the production of a drug from a cultured microbial cell , the improvement consisting in the use , as said microbial cell , of a or related bacterium.3Deinococcus bacterium.. The method of claim 1 , wherein the bacterium is a4Deinococcus bacterium.. The method of claim 1 , wherein the bacterium is a wild-type5Deinococcus bacterium.. The method of claim 1 , wherein the bacterium is a recombinant6. The method of claim 1 , wherein the bacterium is cultured in a defined medium.7. The method of claim 1 , wherein the bacterium is cultured in a complex medium.8. The method of claim 1 , wherein the bacterium is cultured at a pH comprised between 4 and 8.9. The method of claim 1 , wherein the bacterium is cultured in a fermentation device or as a continuous culture.10. The method of claim 1 , wherein the drug is selected from an antibiotic claim 1 , a bacteriostatic agent claim 1 , an anti-parasitic agent claim 1 , an anti-fungal agent claim 1 , an anti-viral agent claim 1 , an anti-metabolite agent claim 1 , a chemotherapeutic agent claim 1 , a cytokine claim 1 , a cell- ...

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13-02-2014 дата публикации

TARGETING POLY-GAMMA-GLUTAMIC ACID TO TREAT STAPHYLOCOCCUS EPIDERMIDIS AND RELATED INFECTIONS

Номер: US20140045213A1

Immunogenic compositions and methods for eliciting an immune response against and other related staphylococci are provided. The immunogenic compositions can include immunogenic conjugates of poly-γ-glutamic acid (such as γDLPGA) polypeptides of , or related staphylococci that express a γPGA polypeptide. The γPGA conjugates elicit an effective immune response against , or other staphylococci, in subjects to which the conjugates are administered. A method of treating an infection caused by a organism that expresses cap genes is also disclosed. The method can include selecting a subject who is at risk of or has been diagnosed with the infection by the organism which expresses γPGA from the cap genes. Further, the expression of a γPGA polypeptide by the organism can then be altered. 1. A method of producing poly-γ-glutamic acid (γPGA) , comprising:{'i': 'Staphylococcus epidermidis', 'culturing one or more strains of under conditions sufficient for producing γPGA.'}2. The method of claim 2 , wherein the method is used to produce poly-γ-DL-glutamic acid (γDLPGA).3Staphylococcus epidermidisS. capitis, S. warneri, S. saccharolyticus, S. caprae, S. hominis, S. haemolyticus. The method of claim 1 , wherein the one or more strains of comprises: or a combination thereof.4Staphylococcus epidermidisS. capitis, S. warneri, S. saccharolyticus, S. caprae, S. hominis, S. haemolyticus. The method of claim 2 , wherein the one or more strains of comprises: or a combination thereof. This is a divisional application of co-pending U.S. application Ser. No. 13/554,245, filed Jul. 20, 2012, which is a divisional application of U.S. application Ser. No. 13/035,716, filed Feb. 25, 2011, now U.S. Pat. No. 8,252,550, which is a divisional of U.S. application Ser. No. 11/994,984, filed Jan. 7, 2008 (now abandoned), which is the §371 U.S. National Stage of International Application No. PCT/US2006/026900, filed Jul. 10, 2006, which was published in English under PCT Article 21(2), which in turn ...

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20-02-2014 дата публикации

METHOD FOR LYSING CELLS

Номер: US20140051088A1
Принадлежит: Merck Patent GmBH

The present invention relates to a method for the lysis of cells, especially bacterial cells and the isolation of nucleic acids. The sample is treated with lysis solution that comprises at least one liquid that is not miscible with water and heated. Afterwards the mixture is cooled down and water is added so that after cooling a two phase system is generated. The nucleic acids can be found in the aqueous phase. 1. A method for lysing cells bya) providing a sample comprising the cellsb) adding a lysis solution at least comprising a water-immiscible liquid to the samplec) heating the mixture obtained in step b) to a temperature above 80° C.d) in case the temperature in step c) was above 100° C., cooling the sample to a temperature below 100° C.e) adding water or an aqueous buffer solution to the mixture, thereby obtaining a water-immiscible liquid phase and an aqueous phase2. Method according to characterized in that the cells are bacterial cells.3. Method according to claim 1 , characterized in that in step c) claim 1 , the mixture is heated to a temperature between 100 and 150° C.4. Method according to claim 1 , characterized in that in step d) claim 1 , the sample is cooled to a temperature between 20 and 40° C.5. Method according to claim 1 , characterized in that the lysis solution added in step b) comprises a water-immiscible ionic liquid.6. Method according to claim 5 , characterized in that the anion of the ionic liquid is bis(trifluormethylsulfonyl)imide [Ntf].7. Method according to claim 1 , characterized in that the lysis solution added in step b) comprises Trihexl(tetradecyl)phosphoniumtris(pentafluorethyl)trifluorophosphate [Ttp][Fap] and/or 1-Butyl-1-methylpyrrolidiniumbis(trifluormethylsulfonyl)imide [bmpyrr][Ntf].8. Method according to claim 1 , characterized in that prior to adding the lysis solution claim 1 , in a step b1) the sample is preincubated with at least one substance comprising an ammonium cation [NR] whereR in each case, independently of ...

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27-02-2014 дата публикации

EVOLUTION OF BOND-FORMING ENZYMES

Номер: US20140057317A1

Strategies, systems, methods, reagents, and kits for the directed evolution of bond-forming enzymes are provided herein. Evolution products, for example, evolved sortases exhibiting enhanced reaction kinetics and/or altered substrate preferences are also provided herein, as are methods for using such evolved bond-forming enzymes. Kits comprising materials, reagents, and cells for carrying out the directed evolution methods described herein are also provided. 1S. aureus. A sortase comprising an amino acid sequence that is at least 90% homologous to the amino acid sequence of Sortase A as provided as SEQ ID NO: 1 or a fragment thereof , wherein the amino acid sequence of the sortase comprises one or more mutations selected from the group consisting of P94S , P94R , E106G , F122Y , F154R , D160N , D165A , G174S , K190E , and K196T.2. The sortase of claim 1 , wherein the sortase comprises an amino acid sequence that is at least 95% claim 1 , at least 98% claim 1 , or at least 99% homologous to SEQ ID NO: 1 or a fragment thereof.3S. aureus. The sortase of claim 1 , wherein the amino acid sequence of the sortase comprises at least one mutation claim 1 , at least two mutations claim 1 , at least three mutations claim 1 , or at least four mutations as compared to the amino acid sequence of Sortase A provided as SEQ ID NO: 1 or a fragment thereof.4. The sortase of claim 1 , wherein the sortase comprises a P94S or P94R mutation claim 1 , a D160N mutation claim 1 , a D165A mutation claim 1 , a K190E mutation claim 1 , and a K196T mutation.5. The sortase of claim 1 , wherein the sortase comprises a P94S or P94R mutation claim 1 , a D160N mutation claim 1 , and a K196T mutation.6. The sortase of claim 1 , wherein the sortase comprises a P94S or P94R mutation claim 1 , a D160N mutation claim 1 , and a D165A mutation.7. The sortase of claim 1 , wherein the sortase comprises a P94S or P94R mutation claim 1 , a D160N mutation claim 1 , a D165A mutation claim 1 , and a K196T mutation ...

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06-03-2014 дата публикации

METHODS FOR RECOVERING PEPTIDES/AMINO ACIDS AND OIL/FAT FROM ONE OR MORE ...

Номер: US20140066359A1
Автор: CARLSSON Tomas
Принадлежит: Zymtech Production AS

According to a first aspect, hydrolysis of a protein-containing raw material and separation of amino acids/peptides is carried out, wherein the hydrolysis is effected by using the endogenous enzymes of the protein-containing raw material. The hydrolysate is passed through a membrane filter, wherein peptide/amino acids follow a permeate stream, whilst the active enzymes continuously break down any protein residues that are deposited on the membrane surface. The enzymes are passed together with retentate back to the hydrolysis. Furthermore, an amino acid and peptide product and an oil product are described and the use thereof is disclosed. 1. A method for producing peptides/amino acids with a fat content of less than 0.5% by weight from a protein-containing raw material , wherein the method comprises:(a) grinding the protein-containing raw material;(b) heating the ground raw material to temperatures in the range of 40-62° C.;(c) adding water which has approximately the same, or the same, temperature as the raw material, and wherein the pH of the water is adjusted to 7.0-8.5;(d) hydrolysing the ground raw material with endogenous enzymes in order to prepare a hydrolysate;(e) removing solid particles and non-hydrolysed proteins which can be returned to the hydrolysis from the hydrolysate;(f) periodically or continually separating off fat/oil in order to obtain an oil product;(g) separating off the desired molecular weight fraction of peptides/amino acids by membrane filtration;(h) routing the portions of the hydrolysate that do not penetrate the membrane filter in step (g) back to the hydrolysis in step (d);(i) concentrating and optionally drying the permeate in order to obtain a distillate comprising water, and a concentrate comprising peptides/amino acids and water; and(j) wholly or partly returning the distillate from the concentrating to the permeate side of the membrane filter.2. The method according to claim 1 , wherein the method takes place as a closed process.3 ...

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13-03-2014 дата публикации

PROTEIN COMPLEXES AND METHODS OF MANUFACTURING BISPECIFIC ANTIBODIES USING THE PROTEIN COMPLEXES

Номер: US20140073767A1
Принадлежит:

A protein complex comprising (i) a first fusion protein comprising (a) a first polypeptide that comprises a first antigen-binding site and (b) a first binding protein linked to a terminus of the first polypeptide; and (ii) a second fusion protein comprising (a) a second polypeptide that comprises a second antigen-binding site and (b) a second binding protein linked to a terminus of the second polypeptide, wherein the protein complex comprises an amino acid sequence that enables cleavage between the first polypeptide and the first binding protein, and/or between the second polypeptide and the second binding protein, as well as a method for preparing a bi-specific antibody and related methods and compositions. 1. A protein complex comprising:(i) a first fusion protein comprising (a) a first polypeptide that comprises a first antigen-binding site and (b) a first binding protein linked to a terminus of the first polypeptide; and(ii) a second fusion protein comprising (a) a second polypeptide that comprises a second antigen-binding site and (b) a second binding protein linked to a terminus of the second polypeptide,wherein the protein complex comprises an amino acid sequence that enables cleavage between the first polypeptide and the first binding protein, or between the second polypeptide and the second binding protein, or wherein the protein complex comprises both an amino acid sequence that enables cleavage between the first polypeptide and the first binding protein, and an amino acid sequence that enables cleavage between the second polypeptide and the second binding protein.2. The protein complex of claim 1 , wherein the first antigen-binding site and the second antigen-binding site are positioned at the N-terminus of the first polypeptide and the second polypeptide claim 1 , respectively.3. The protein complex of claim 1 , wherein an antigen specifically binding to the first antigen-binding site and an antigen specifically binding to the second antigen-binding site ...

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20-03-2014 дата публикации

ACID-CLEAVABLE LINKERS EXHIBITING ALTERED RATES OF ACID HYDROLYSIS

Номер: US20140080998A1
Принадлежит: E I DU PONT DE NEMOURS AND COMPANY

An acid-cleavable peptide linker comprising aspartic acid and proline residues is disclosed. The acid-cleavable peptide linker provides an altered sensitivity to acid-hydrolytic release of peptides of interest from fusion peptides of the formula PEP1-L-PEP2. The inventive linker, L, is described in various embodiments, each of which provides substantially more rapid acid-release of peptides of interest than does a single aspartic acid-proline pair. In an additional aspect, a method of increasing the stability of an acid cleavable linkage to acid hydrolysis is also provided. 1. A method of preparing at least one peptide of interest (“POI”) from a fusion peptide comprising at least one POI , comprising:{'claim-ref': {'@idref': 'CLM-00021', 'claim 21'}, 'a) providing a recombinant cell synthesizing the fusion peptide of ;'}b) contacting the fusion peptide with a solution of sufficiently acidic pH so that linker L is cleaved, andc) isolating the at least one POI.2. (canceled)3. The method of wherein the recombinant cell is a recombinant microbial cell.4. The method of wherein the recombinant microbial cell is a recombinant yeast cell.5. The method of wherein the recombinant microbial cell is a recombinant bacterial cell.6. The method of wherein the acid-cleavable linker is cleaved by incubating the fusion peptides at a pH in the range from about pH 1 to about pH 4.7. The method of wherein the acid-cleavable linker is cleaved by incubating the fusion peptides at a pH in the range from about pH 2 to about pH 4.8. The method of wherein the acid-cleavable linker is cleaved by incubating the fusion peptides at a pH in the range from about pH 3 to about pH 4.9. The method of wherein the acid-cleavable linker is cleaved by incubating the fusion peptides at a pH of about 4.10. The method of wherein the acid-cleavable linker is cleaved by incubating the fusion peptides at a temperature of about 40° C. to about 90° C.11. The method of wherein the acid-cleavable linker is cleaved ...

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27-03-2014 дата публикации

METHODS AND DEVICES FOR SAMPLE LYSIS

Номер: US20140087359A1
Принадлежит: California Institute of Technology

Disclosed herein are methods and systems for use in preparing a sample. The methods and systems may be used for lysing one or more structures in a sample (e.g., cells, viral particles, etc.). The methods and compositions may comprise a microfluidic chip or use thereof. The microfluidic chips disclosed herein may comprise (a) a substrate comprising a chamber, wherein at least one mechanical element may be located within the chamber; (b) a thermal element in contact with the chamber; and (c) at least one aperture within the surface of the substrate, wherein the aperture may be configured to insulate the chamber. 1. A microfluidic chip comprising:a. a chamber, wherein at least one stir bar is located within the chamber;b. a thermal device in thermal contact with the chamber; andc. at least one insulative groove within the surface of the chip, which is configured to insulate the chamber.2. A microfluidic chip comprising:a. a chamber, wherein a plurality of granular particles is located within the chamber;b. a thermal device in thermal contact with the chamber; andc. at least one insulative groove within the surface of the chip, which is configured to insulate the chamber.3. The microfluidic chip of or , wherein the thermal device is a heater.4. The microfluidic chip of or , wherein the thermal device is a cooler.5. The microfluidic chip of or , wherein the at least one insulative groove is nine insulative grooves.6. The microfluidic chip of or , wherein the at least one insulative groove is a plurality of grooves positioned around the chamber.7. The microfluidic chip of or , wherein the at least one insulative groove is filled with an insulative material.8. The microfluidic chip of claim 1 , wherein the stir bar is composed of magnetic material.9. The microfluidic chip of or claim 1 , wherein the microfluidic chip is coupled to an external magnetic field.10. The microfluidic chip of claim 2 , wherein the granular particles are beads.11. The microfluidic chip of claim 10 ...

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07-01-2016 дата публикации

FORMULATIONS AND METHODS FOR INCREASED RECOMBINANT PROTEIN PRODUCTION

Номер: US20160002592A1
Принадлежит:

Formulations and methods to increase the production of recombinant proteins, and other aspects, are disclosed. The formulations and methods relate to increasing mannose or calcium concentration, or both, in a cell culture medium formulation for culturing cells that express recombinant proteins. In some embodiments, a mammalian cell culture medium formulation is provided that has at least one of mannose at about 3.5 g/L or more and calcium in a range from about 1.5 mM to about 9.5 mM. Numerous other aspects and/or embodiments are provided. 1. A mammalian cell culture medium formulation comprising at least one of mannose at about 3.5 g/L or more and calcium in a range from about 1.5 mM to about 9.5 mM.2. The formulation of claim 1 , wherein the medium comprises mannose at about 3.5 g/L or more and calcium at less than about 1.5 mM or more than about 9.5 mM.3. The formulation of claim 1 , wherein the medium comprises mannose at less than about 3.5 g/L and calcium in a range from about 1.5 mM to about 9.5 mM.4. The formulation of claim 1 , wherein the medium comprises mannose in a range from about 4 g/L to about 5 g/L.5. The formulation of claim 2 , wherein the medium comprises mannose in a range from about 4 g/L to about 5 g/L.6. The formulation of claim 1 , wherein the medium comprises calcium in a range from about 2 mM to about 5 mM.7. The formulation of claim 3 , wherein the medium comprises calcium in a range from about 2 mM to about 5 mM.8. The formulation of claim 1 , wherein the formulation comprises DMEM/F12 in 1:1 ratio claim 1 , and includes at least one of mannose in a range from about 4 g/L to about 5 g/L and calcium in a range from about 2 mM to about 5 mM.9. A method of producing a recombinant protein in cell culture claim 1 , comprising:providing recombinant protein expressing cells; andculturing the cells in a cell culture medium including at least one of mannose at about 3.5 g/L or more and calcium in a range from about 1.5 mM to about 9.5 mM.10. The ...

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02-01-2020 дата публикации

Mating Factor Alpha Pro-Peptide Variants

Номер: US20200002388A1
Автор: Noergaard Per
Принадлежит:

The present invention is related to Mating Factor α pro-peptide variants useful for the recombinant expression of polypeptides comprising a GLP-1 peptide in yeasts. The invention is also related to DNA sequences, vectors and host cells for use in expressing polypeptides in yeasts. 2. The Mating Factor α pro-peptide variant according to wherein{'sub': '38', 'Xis V,'}{'sub': '39', 'Xis selected from L, I, V or M,'}{'sub': '40', 'Xis selected from G, R or K,'}{'sub': '41', 'Xis Y, and'}{'sub': '42', 'Xis selected from L or I.'}3. The Mating Factor α pro-peptide variant according to claim 1 , wherein{'sub': '38', 'Xis V,'}{'sub': '39', 'Xis selected from L, I, V or M,'}{'sub': '40', 'Xis selected from G or R,'}{'sub': '41', 'Xis Y, and'}{'sub': '42', 'Xis L.'}4. The Mating Factor α pro-peptide variant according to claim 3 , wherein Xis R.5. The Mating Factor α pro-peptide variant according to claim 1 , wherein three of the amino acid residues in X-X-X-X-Xare identical to the corresponding amino acid residues in VIGYL (SEQ ID NO:3).7. An expression vector comprising a DNA sequence encoding a polypeptide according to .8. A host cell comprising the expression vector according to .10. The method according to claim 9 , wherein Xis R.11. The method according to claim 9 , wherein said Mating Factor α pro-peptide variant has less than 10 amino acid residue changes outside of the X-Xsequence as compared to the Mating Factor α pro-peptide as set out in SEQ ID NO:2 (amino acid residues 20-85).12. The method according to claim 9 , wherein said yeast carries at least one genetic modification reducing its capacity for O-glycosylation.13. The method according to claim 9 , wherein the PMT1 gene in said yeast is deleted.14. The method according to claim 9 , wherein said polypeptide comprises GLP-1(9-37)[K34R] or GLP-1(9-37)[K34R claim 9 ,G37K].15. The method according to claim 9 , wherein said polypeptide consists of GLP-1(9-37)[K34R] or GLP-1(9-37)[K34R claim 9 ,G37K].16. The method ...

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07-01-2021 дата публикации

BIOREACTIVE COMPOSITIONS AND METHODS OF USE THEREOF

Номер: US20210002325A1
Автор: Wang Lei, Wang Nanxi
Принадлежит:

Provided herein are, inter alia, non-toxic, bioreactive unnatural amino acids and methods of using same. 3. The biomolecule conjugate of claim 2 , wherein{'sup': 1', '3A', '3A', '3A', '3A', '3B, 'sub': '2', 'Lis a bond, —S(O)—, —NR—, —O—, —S—, —C(O)—, —C(O)NR—, —NRC(O)—, —NRC(O)NR—, —C(O)O—, —OC(O)—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene;'}{'sup': 2', '4A', '4A', '4A', '4A', '4B, 'sub': '2', 'Lis a bond, —S(O)—, —NR—, —O—, —S—, —C(O)—, —C(O)NR—, —NRC(O)—, —NRC(O)NR—, —C(O)O—, —OC(O)—, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, or substituted or unsubstituted heteroarylene; and'}{'sup': 3A', '3B', '4A', '4B, 'R, R, R, and Rare independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.'}4. The biomolecule conjugate of claim 2 , wherein Xis —NH— claim 2 , —O— claim 2 , or imidazolylene.5. The biomolecule conjugate of claim 1 , wherein the first biomolecule moiety is a peptidyl moiety claim 1 , a nucleic acid moiety claim 1 , or a carbohydrate moiety.6. The biomolecule conjugate of claim 5 , wherein the first biomolecule moiety is a peptidyl moiety; and wherein the peptidyl moiety is covalently bonded to the bioconjugate linker via lysine claim 5 , histidine claim 5 , or tyrosine.7. The biomolecule conjugate of claim 1 , wherein the second biomolecule moiety is a peptidyl moiety claim 1 , a nucleic acid moiety claim 1 , or a carbohydrate moiety.8. The biomolecule conjugate ...

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07-01-2021 дата публикации

MUTANT TYPE 2-DEOXY-SCYLLO-INOSOSE SYNTHASE

Номер: US20210002687A1
Принадлежит:

A polypeptide includes, in the amino acid sequence of SEQ ID NO: 1 or a similar sequence, at least one specific amino acid substitution on at least one of the 14th, 37th, 209th, 293rd, and 319th amino acid residues from the N-terminal of the amino acid sequence of SEQ ID NO: 1. A polynucleotide, an expression cassette, a vector, and a transformant include a base sequence encoding the amino acid sequence of the polypeptide. A method of producing the polypeptide and a method of producing 2-deoxy-scyllo-inosose are also provided. 1. A polypeptide comprising at least one amino acid mutation selected from the group consisting of the following (a) to (e) in an amino acid sequence of the following (A1) or (A2):(A1) an amino acid sequence of SEQ ID NO: 1;(A2) an amino acid sequence of a polypeptide having an enzymatic activity that produces 2-deoxy-scyllo-inosose from glucose 6-phosphate, the amino acid sequence (A2) having a sequence identity of 80% or higher with the amino acid sequence of SEQ ID NO: 1,(a) an amino acid mutation in which an amino acid residue corresponding, in terms of alignment, to an asparagine residue that is a 14th amino acid residue from the N-terminal in the amino acid sequence of SEQ ID NO: 1 is substituted with threonine;(b) an amino acid mutation in which an amino acid residue corresponding, in terms of alignment, to a tyrosine residue that is a 37th amino acid residue from the N-terminal in the amino acid sequence of SEQ ID NO: 1 is substituted with phenylalanine;(c) an amino acid mutation in which an amino acid residue corresponding, in terms of alignment, to an alanine residue that is a 290th amino acid residue from the N-terminal in the amino acid sequence of SEQ ID NO: 1 is substituted with threonine;(d) an amino acid mutation in which an amino acid residue corresponding, in terms of alignment, to a tryptophan residue that is a 293rd amino acid residue from the N-terminal in the amino acid sequence of SEQ ID NO: 1 is substituted with ...

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03-01-2019 дата публикации

RECOMBINANT PRODUCTION METHOD

Номер: US20190002947A1
Принадлежит: Hoffmann-La Roche Inc.

Herein is reported a method for the recombinant production of a polypeptide in a eukaryotic cell comprising the steps of (i) cultivating a eukaryotic cell comprising a nucleic acid encoding the polypeptide in a cultivation medium comprising a compound selected from the group consisting of trans-2-methyl 2-pentenoic acid, the broad-spectrum HDAC inhibitor Quisinostat, and the subtype-specific HDAC inhibitor Romidepsin, and (ii) recovering the polypeptide from the cell or the cultivation medium and thereby producing the polypeptide in a eukaryotic cell. 1. A method for the recombinant production of a polypeptide in a eukaryotic cell comprising the steps of:cultivating a eukaryotic cell comprising a nucleic acid encoding the polypeptide in a cultivation medium comprising a compound selected from the group consisting of trans-2-methyl 2-pentenoic acid, the broad-spectrum HDAC inhibitor Quisinostat, and the subtype-specific HDAC inhibitor Romidepsin, andrecovering the polypeptide from the cell or the cultivation medium and thereby producing the polypeptide in a eukaryotic cell.2. A method for the recombinant production of a polypeptide in a eukaryotic cell comprising the steps of:cultivating a eukaryotic cell comprising a nucleic acid encoding the polypeptide in a cultivation medium comprising a first and a second compound selected from the group of compounds consisting of trans-2-methyl 2-pentenoic acid, subtype-specific HDAC inhibitors, and canonical and non-canonical NFκB activators, andrecovering the polypeptide from the cell or the cultivation medium and thereby producing the polypeptide in a eukaryotic cell.3. The method according to claim 2 , wherein the canonical NFκB activator is selected from the group consisting of TNFalpha claim 2 , ligands of IL-1R/TLR claim 2 , lipopolysaccharides and IL-1b.4. The method according to claim 2 , wherein the non-canonical NFκB activator is selected from the group consisting of EGF claim 2 , PMA claim 2 , and betulinic acid.5. ...

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01-01-2015 дата публикации

CELLS AND METHODS FOR PRODUCING LUTEIN

Номер: US20150005534A1
Принадлежит:

Provided herein are recombinant cells (e.g., recombinant bacteria or plant, insect, mammalian, and yeast cells) containing a nucleic acid encoding a CYP97A protein or a nucleic acid encoding a CYP97B protein; a nucleic acid encoding a CYP97C protein; a nucleic acid encoding a geranylgeranyl pyrophosphate synthase protein; a nucleic acid encoding a phytoene synthase protein; a nucleic acid encoding a phytoene desaturase protein; a nucleic acid encoding a lycopene β-cyclase protein; and a nucleic acid encoding a lycopene ε-cyclase protein. Also provided are methods of producing lutein that include culturing these recombinant cells (e.g., recombinant bacteria and yeast cells), and methods of generating these recombinant cells (e.g., recombinant bacteria and yeast cells). Also provided is lutein produced by these methods, and pharmaceutical compositions, food supplements, food products, and cosmetic compositions that contain lutein produced by these methods. 1. A recombinant bacterium or yeast cell comprising:a nucleic acid encoding a CYP97A protein or a nucleic acid encoding a CYP97B protein;a nucleic acid encoding a CYP97C protein;a nucleic acid encoding a geranylgeranyl pyrophosphate synthase protein;a nucleic acid encoding a phytoene synthase protein;a nucleic acid encoding a phytoene desaturase protein;a nucleic acid encoding a lycopene β-cyclase protein; anda nucleic acid encoding a lycopene ε-cyclase protein.2. The recombinant bacterium or yeast cell of claim 1 , further comprising:a nucleic acid encoding a D-1-deoxyxylulose 5-phosphate synthase protein; and/ora nucleic acid encoding an isopentenyl pyrophosphate isomerase protein.3. The recombinant bacterium or yeast cell of claim 1 , wherein the bacterium or yeast cell comprises a nucleic acid encoding a CYP97A protein.4. The recombinant bacterium or yeast cell of claim 3 , wherein the CYP97A protein comprises a sequence at least 80% identical to SEQ ID NO: 1.5. The recombinant bacterium or yeast cell of claim 1 ...

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12-01-2017 дата публикации

ENHANCEMENT OF RECOMBINANT PROTEIN EXPRESSION USING A MEMBRANE-BASED CELL RETENTION SYSTEM

Номер: US20170009269A1
Автор: OZTURK Sadettin Seyit
Принадлежит:

The invention disclosed herein provides a novel use of an external membrane-based cell retention system in conjunction with perfusion cell culture for improved cell expression of recombinant proteins, particularly coagulation proteins such as rFVIII, B-Domain Deleted rFVIII, rFIX or rFVII/rFVIIa. The use of such a system at high cell density results in a more homogeneous cell culture due to mechanical forces induced during the operation of the retention system, such as the cell circulation induced by pumping through the fibers. 1. A method of increasing cell expression of mammalian cells comprising a membrane-based external cell retention system used in conjunction with a perfusion cell culture.2. The method of claim 1 , wherein the method of increasing cell expression of mammalian cells is used to produce recombinant proteins.3. The method of claim 2 , wherein the recombinant proteins expressed are coagulation proteins.4. The method of claim 3 , wherein the expressed coagulation proteins are chosen from the group consisting of recombinant Factor IX claim 3 , recombinant FVIII claim 3 , B-Domain-Deleted recombinant Factor FVIII claim 3 , recombinant VII and recombinant Factor VIIa.5. The method of claim 1 , wherein the mammalian cells are chosen from CHO claim 1 , BHK and mammalian cells.6. The method of claim 5 , wherein the membrane based external cell retention system used in conjunction with perfusion cell culture is capable of expressing B-Domain Deleted recombinant Factor VIII and provides an increase in production of B-Domain Deleted recombinant FVIII protein of at least 50% over that obtained with a cell retention system not utilizing a membrane filtration system.7. The method of claim 1 , wherein the perfusion cell culture is performed in a disposable bioreactor.8. The method of claim 1 , wherein the membrane-based cell retention system is disposable.9. The method of claim 1 , wherein the cell culture system is disposable because both the bioreactor that ...

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14-01-2016 дата публикации

Composition for synthesizing protein with reduced lipopolysaccharide contamination, method for producing protein using said composition

Номер: US20160010079A1
Принадлежит: Genefrontier Corp, University of Tokyo NUC

According to the present invention, a composition possessing cell-free protein synthesis activity with reduced contaminating lipopolysaccharide, and a method for producing a protein using the same are provided. When ribosome display is performed using the composition and method for protein production of the present invention, the background that is caused by non-specific binding is reduced, so that a nucleic acid that encodes the desired polypeptide can be selected with high accuracy and high efficiency.

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11-01-2018 дата публикации

FORMULATIONS AND METHODS FOR INCREASED RECOMBINANT PROTEIN PRODUCTION

Номер: US20180010090A1
Принадлежит:

Formulations and methods to increase the production of recombinant proteins, and other aspects, are disclosed. The formulations and methods relate to increasing mannose or calcium concentration, or both, in a cell culture medium formulation for culturing cells that express recombinant proteins. In some embodiments, a mammalian cell culture medium formulation is provided that has at least one of mannose at about 3.5 g/L or more and calcium in a range from about 1.5 mM to about 9.5 mM. Numerous other aspects and/or embodiments are provided. 1. A mammalian cell culture medium formulation comprising at least one of mannose at about 3.5 g/L or more and calcium in a range from about 1.5 mM to about 9.5 mM.2. The formulation of claim 1 , wherein the medium comprises mannose at about 3.5 g/L or more and calcium at less than about 1.5 mM or more than about 9.5 mM.3. The formulation of claim 1 , wherein the medium comprises mannose at less than about 3.5 g/L and calcium in a range from about 1.5 mM to about 9.5 mM.4. The formulation of claim 1 , wherein the medium comprises mannose in a range from about 4 g/L to about 5 g/L.5. The formulation of claim 2 , wherein the medium comprises mannose in a range from about 4 g/L to about 5 g/L.6. The formulation of claim 1 , wherein the medium comprises calcium in a range from about 2 mM to about 5 mM.7. The formulation of claim 3 , wherein the medium comprises calcium in a range from about 2 mM to about 5 mM.8. The formulation of claim 1 , wherein the formulation comprises DMEM/F12 in 1:1 ratio claim 1 , and includes at least one of mannose in a range from about 4 g/L to about 5 g/L and calcium in a range from about 2 mM to about 5 mM.9. A method of producing a recombinant protein in cell culture claim 1 , comprising:providing recombinant protein expressing cells; andculturing the cells in a cell culture medium including at least one of mannose at about 3.5 g/L or more and calcium in a range from about 1.5 mM to about 9.5 mM.10. The ...

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11-01-2018 дата публикации

A CRISPR-CAS SYSTEM FOR A YEAST HOST CELL

Номер: US20180010151A1
Принадлежит:

The present invention relates to the field of molecular biology and cell biology. More specifically, the present invention relates to a CRISPR-CAS system for a yeast host cell. 1SaccharomycesKluyveromyces. A non-naturally occurring or engineered composition comprising a source of a CRISPR-Cas system comprising a guide-polynucleotide and a Cas protein , wherein the guide-polynucleotide comprises a guide-sequence that essentially is the reverse complement of a target-polynucleotide in a host cell and the guide-polynucleotide can direct binding of the Cas protein at the target-polynucleotide in the host cell to form a CRISPR-Cas complex , wherein the guide-sequence is essentially the reverse complement of the (N)y part of a 5′-(N)yPAM-3′ polynucleotide sequence target in the genome of the host cell , wherein y is an integer of 8-30 , wherein PAM is a protospacer adjacent motif , wherein the host cell is a eukaryote , which eukaryote is a yeast , optionally a or a and wherein PAM is optionally a sequence selected from the group consisting of 5′-XGG-3′ , 5′-XGGXG-3′ , 5′-XXAGAAW-3′ , 5′-XXXXGATT-3′ , 5′-XXAGAA-3′ , 5′-XAAAAC-3′ , wherein X can be any nucleotide or analog thereof , optionally X can be any nucleotide; and W is A or T.2. A composition according to claim 1 , wherein the Cas protein is encoded by a polynucleotide and/or the guide-polynucleotide is encoded by or present on a polynucleotide.3. A composition according to claim 1 , wherein the Cas protein is encoded by a polynucleotide and/or the guide-polynucleotide is encoded by or present on another polynucleotide and the polynucleotide or polynucleotides are comprised in a vector.4. A composition according to claim 1 , wherein the guide polynucleotide is encoded by a polynucleotide that is transcribed to provide for the actual guide-polynucleotide.5. A composition according to claim 1 , wherein a polynucleotide encoding a guide-polynucleotide has sequence identity with a vector such that recombination of the ...

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11-01-2018 дата публикации

METHOD OF CONTINUOUSLY PRODUCING GLUTATHIONE USING PHOTOSYNTHETIC MEMBRANE VESICLES

Номер: US20180010163A1
Принадлежит: SOGANG UNIVERSITY RESEARCH FOUNDATION

The present invention relates to a method of producing glutathione, wherein photosynthetic membrane vesicles and enzymes catalyzing glutathione synthesis are combined and glutamate, cysteine and glycine are used as reaction substrates. As enzymes catalyzing glutathione synthesis, γ-glutamylcysteine synthetase and glutathione synthetase may be used together, or bifunctional glutathione synthetase may be used alone. According to the conventional methods, there is a problem in that expensive adenosine triphosphate should be continuously supplied when glutathione is produced. However, according to the present invention, since photosynthetic membrane vesicles are used as a source to regenerate adenosine triphosphate, it is possible to continuously produce glutathione without additionally adding adenosine triphosphate, thereby reducing production costs of glutathione. 1. A method of producing glutathione , comprising:a) a step of generating adenosine triphosphate (ATP) from adenosine diphosphate (ADP) and an inorganic phosphate by irradiating photosynthetic membrane vesicles with light; andb) a step of synthesizing glutathione by enzymes that catalyze glutathione synthesis using ATP generated in step a), and forming ADP and an inorganic phosphate.2. The method according to claim 1 , further comprising c) a step of reusing ADP and an inorganic phosphate forming in step b) to generate ATP in photosynthetic membrane vesicles.3. The method according to claim 1 , wherein the photosynthetic membrane vesicles are selected from the group consisting of chromatophore membrane vesicles isolated from purple non-sulfur bacteria and thylakoid membrane vesicles isolated from cyanobacteria or algae.4. The method according to claim 1 , wherein the enzymes that catalyze glutathione synthesis are one or more selected from the group consisting of γ-glutamylcysteine synthetase (GSH-I) claim 1 , glutathione synthetase (GSH-II) and bifunctional glutathione synthetase (bifunctional γ- ...

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14-01-2021 дата публикации

METHOD FOR PREPARING ACTIVE PROTEIN HYDROLYSATES BY HYDROLYZING CEREAL PROTEINS WITH MALT TOGETHER WITH PROTEASES

Номер: US20210010047A1
Принадлежит:

This disclosure relates to the technological field of nutrient food, particularly relates to a method for preparing active protein hydrolysates by hydrolyzing cereal proteins with malt together with proteases, including the following steps: (1) grinding the malt, obtaining the malt flour; (2) hydrolyzing cereal proteins with the malt flour together with proteases to prepare active protein hydrolysates. The present invention uses the malt flour together with proteases to hydrolyze cereal proteins, which can effectively reduce the consumption of proteases and save the cost, moreover, aminopeptidases and carboxypeptidases in the malt are able to hydrolyze peptide linkages from the ends of proteins, and the bitter hydrophobic amino acids are cut off, thus effectively decreasing the bitterness of the products, on the other hand, which can increase the additional values of the malt, providing its new applications. 1. A method for preparing active protein hydrolysates by hydrolyzing cereal proteins with malt together with proteases , which is characterized in that: including the following steps:(1) grinding the malt, obtaining the malt flour;(2) hydrolyzing cereal proteins with the malt flour together with proteases to prepare active protein hydrolysates.2. The method for preparing active protein hydrolysates by hydrolyzing cereal proteins with malt together with proteases of claim 1 , which is characterized in that: the step (2) was specifically: grinding dry cereal proteins into powders claim 1 , which were prepared into a suspension of cereal proteins claim 1 , hydrolyzed by adding the malt flour first claim 1 , then hydrolyzed by adding proteases claim 1 , upon which they were dried.3. The method for preparing active protein hydrolysates by hydrolyzing cereal proteins with malt together with proteases of claim 2 , which is characterized in that: the substrate concentration of cereal proteins in the step (2) was 5-15% (w/v).4. The method for preparing active protein ...

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14-01-2021 дата публикации

METHOD FOR PRODUCING ASTAXANTHIN

Номер: US20210010049A1
Принадлежит:

Efficiency of production method for producing astaxanthin by culturing microalgae is improved. A method for producing astaxanthin in which astaxanthin is produced in inner of algae by culturing a microalga, wherein a light irradiation during a green stage culturing of the microalga is performed by using both a blue LED of peak wavelength from 420 to 500 nm and a red LED of peak wavelength from 620 to 690 nm, and having a ratio of photon flux density of the blue LED to the red LED to be 2:3 to 20:1. It is preferable that the photon flux density of the blue LED is from 5 to 200 μmol/m/s, and the photon flux density of the red LED is from 5 to 200 μmol/m/s. 1. A method for producing astaxanthin in which astaxanthin is produced in inner of algae by culturing a microalgae , wherein a light irradiation during a green stage culturing of the microalgae is performed by using both a blue LED of peak wavelength from 420 to 500 nm and a red LED of peak wavelength from 620 to 690 nm , and having a ratio of photon flux density of the blue LED to the red LED to be 2:3 to 20:1.2. The method for producing astaxanthin according to claim 1 , wherein the ratio of photon flux density of the blue LED to the red LED is 3:2 to 20:1.3. The method for producing astaxanthin according to claim 1 , wherein the light irradiation using the blue LED and the red LED is continuously performed during the green stage culturing.4. The method for producing astaxanthin according to claim 1 , wherein the photon flux density of the blue LED is from 5 to 200 μmol/m/s claim 1 , and the photon flux density of the red LED is from 5 to 200 μmol/m/s.5. The method for producing astaxanthin according to claim 1 , wherein a light irradiation during a red stage culturing of the microalgae is performed further using both the blue LED of peak wavelength from 420 to 500 nm and the red LED of peak wavelength from 620 to 690 nm.6. The method for producing astaxanthin according to claim 5 , wherein the ratio of photon ...

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11-01-2018 дата публикации

IDENTIFICATION OF THE PRESENCE OF SPECIFIC POLYPEPTIDES BY LIQUID CHROMATOGRAPHY AND MASS SPECTROMETRY

Номер: US20180011107A1
Принадлежит: Genysis Lab Inc.

Disclosed are methods for determining the presence of one or more proteins in a sample, the methods comprising: enzymatically digesting the sample with a protease activity to generate a plurality of proteolytic peptides; separating the plurality of proteolytic peptides using liquid chromatography; performing mass spectrometry on the separated plurality of peptides; and wherein a protein is present in the sample when three or more target peptides for the protein are present among the proteolytic peptides; and wherein the target peptides are selected from the groups consisting of SEQ ID NOS:6-8, SEQ ID NOS:9-11, SEQ ID NOS:12-14, SEQ ID NOS:15-17, and SEQ ID NOS:18-20. In embodiments, a known quantity of a standard peptide may be added to the proteolytic peptides. 1. A method for determining the presence of one or more proteins in a sample , the method comprising:enzymatically digesting the sample with a protease activity to generate a plurality of proteolytic peptides;separating the plurality of proteolytic peptides using liquid chromatography;performing mass spectrometry on the separated plurality of peptides; andwherein a protein is present in the sample when three or more target peptides for the protein are present among the proteolytic peptides; andwherein the target peptides are selected from the groups consisting of SEQ ID NOS:6-8, SEQ ID NOS:9-11, SEQ ID NOS:12-14, SEQ ID NOS:15-17, and SEQ ID NOS:18-20.2. The method according to claim 1 , further comprising adding a known quantity of a standard peptide to the proteolytic peptides.3. The method according to claim 2 , wherein the standard peptide is β-Casomorphin 1-4.4. The method according to claim 1 , wherein the liquid chromatograph is high-performance liquid chromatography.5. The method according to claim 1 , wherein the protease activity is selected from the group consisting of serine proteases claim 1 , trypsin claim 1 , hepsin claim 1 , SCCE claim 1 , TADG12 claim 1 , TADG14 claim 1 , metalloproteases ...

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09-01-2020 дата публикации

Enhanced Production of Recombinant Proteins By Transient Transfection of Suspension-Growing Mammalian Cells

Номер: US20200010808A1
Принадлежит: NATIONAL RESEARCH COUNCIL OF CANADA

Disclosed is a new process for the production of recombinant proteins, by transient transfection of suspension-grown human embryonic kidney cells (293 cell line and its genetic variants) with an expression vector, using polyethylenimine (PEI) as a transfection reagent. In a preferred embodiment, the process uses 293E cells expressing the Epstein-Barr virus (EBV) EBNA 1 protein, in combination with an oriP-based episomal expression vector having an improved cytomegalovirus expression cassette comprising the CMV5 promoter. The process combines in a single step the cell growth, transfection and protein expression, is carried out without changing the culture medium, and allows to achieve high expression levels in a short period of time. The process may be carried out in a serum-free, low-protein culture medium, is easily scalable, compatible with continuous production processes, and fully adapted to high-throughput production of milligram quantities of recombinant proteins. 150-. (canceled)52. The expression vector of claim 51 , wherein the size of the expression vector is from about 4185 base pairs to about 5925 base pairs.53. The expression vector of claim 51 , wherein the fragment of SEQ ID NO: 1 consists of a BxtXI-EcoRI FR fragment consisting of nucleotides 5 to 299 of SEQ ID NO: 1.54. The expression vector of claim 51 , wherein the fragment of SEQ ID NO: 2 consists of a BxtXI FR fragment consisting of nucleotides 300 to 595 of SEQ ID NO: 1.55. The expression vector of claim 51 , further comprising an antibiotic resistance gene and a bacterial origin of replication claim 51 , wherein the antibiotic resistance gene and the bacterial origin of replication are located between SEQ ID NO: 1 or the fragment thereof and the 5′ end of the CMV5 promoter.56. The expression vector of claim 55 , wherein the bacterial origin of replication is pMB1 and/or the antibiotic resistance gene is an ampicillin resistance gene.57. The expression vector of claim 51 , further comprising a ...

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03-02-2022 дата публикации

PEPTIDE MARKERS FOR AUTHENTICATION OF EDIBLE BIRD'S NEST AND RELATED PRODUCTS

Номер: US20220033871A1
Принадлежит:

Provided herein is a method of authenticating using peptide markers found in edible bird's nest hydrolysate. The method can be used to authenticate edible bird's nest and related products and/or distinguish between white edible bird's nest and grass edible bird's nest. 1. A method of identifying an edible bird's nest (EBN) in a sample suspected of comprising the EBN , the method comprising:providing a hydrolysate of the sample;analyzing the hydrolysate using a mass spectroscopy method;determining whether the hydrolysate comprises one or more peptide markers, wherein the one or more peptide markers have an observed mass to charge ratio (m/z) selected from the group consisting of: 277.6012-277.7012 (BNM201), 280.1454-280.2454 (BNM202), 280.6246-280.7246 (BNM203), 292.1148-292.2148 (BNM204), 294.7642-294.8642 (BNM205), 300.0959-300.1959 (BNM206), 305.1166-305.2166 (BNM207), 321.6531-321.7531 (BNM208), 373.7747-373.8747 (BNM209), 381.6542-381.7542 (BNM210), 391.6929-391.7929 (BNM211), 404.1486-404.2486 (BNM212), 410.6669-410.7669 (BNM213), 433.6506-433.7506 (BNM214), 441.6666-441.7666 (BNM215), 454.6518-454.7518 (BNM216), 468.6939-468.7939 (BNM217), 477.6830-477.7830 (BNM218), 498.7467-498.8467 (BNM219), 630.7600-630.8600 (BNM220), 711.3910-711.3910 (BNM221), 820.3134-820.4134 (BNM222), 844.3228-844.4228 (BNM223), 335.1730-335.2730 (BNM224), 417.6510-417.7510 (BNM225), and 447.6496-447.7496 (BNM226); andidentifying based on the whether the hydrolysate comprises the one or more peptide markers if the sample comprises the EBN.2. The method of further comprising the step of hydrolyzing the sample thereby forming the hydrolysate of the sample.3. The method of further comprising the step of hydrolyzing the sample using a protease thereby forming the hydrolysate of the sample.4. The method of claim 3 , wherein the protease is selected from the group consisting of trypsin claim 3 , chymotrypsin claim 3 , lysine protease claim 3 , aspartic protease claim 3 , pepsin claim 3 , ...

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15-01-2015 дата публикации

EFFICIENT PRODUCTION OF PEPTIDES

Номер: US20150017685A1
Принадлежит:

The present invention relates to processes for the production of peptides, and the peptides produced accordingly. Peptides produced according to the invention may be produced more efficiently than peptides produced according to prior art processes. The production process of the invention may lead to advantages in yield, purity, and/or price. Methods of marketing peptides are also disclosed. 122-. (canceled)23. A method for producing a target peptide , the method comprising:expressing a soluble heterologous fusion peptide in a genetically modified cell, the soluble heterologous fusion peptide comprising an affinity tag, a cleavable tag, and the target peptide, wherein the cleavable tag is tryptophan (Trp), and wherein the affinity tag or the cleavable tag is heterologous to the target peptide;binding the soluble heterologous fusion peptide to an affinity material via the affinity tag; andcleaving the soluble heterologous fusion peptide with N-chlorosuccinimide while bound to the affinity material to release the target peptide, thereby producing the target peptide.24. The method of claim 23 , wherein the target peptide is selected from the group consisting of amyloid beta claim 23 , calcitonin claim 23 , enfuvirtide claim 23 , epoetin claim 23 , epoetin delta claim 23 , erythropoietin claim 23 , exenatide claim 23 , factor VIII claim 23 , factor X claim 23 , glucocerebrosidase claim 23 , glucagon-like peptide-1 (GLP-1) claim 23 , granulocyte-colony stimulating factor (G-CSF) claim 23 , human growth hormone (hGH) claim 23 , insulin claim 23 , insulin A claim 23 , insulin B claim 23 , insulin-like growth factor 1 (IGF-1) claim 23 , interferon claim 23 , liraglutide claim 23 , somatostatin claim 23 , teriparatide claim 23 , and tissue plasminogen activator (TPA).25. The method of claim 23 , wherein the step of expressing a soluble heterologous fusion peptide in a genetically modified cell is performed in a bacterial expression system.26E. coli. The method of claim 23 , ...

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19-01-2017 дата публикации

BINDING FUSION PROTEINS, BINDING FUSION PROTEIN-DRUG CONJUGATES, XTEN-DRUG CONJUGATES AND METHODS OF MAKING AND USING SAME

Номер: US20170016042A1
Принадлежит:

The present invention relates to binding fusion protein compositions comprising targeting moieties linked to extended recombinant polypeptide (XTEN), binding fusion protein-drug conjugate compositions, and XTEN-drug conjugate compositions, isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of using such compositions in treatment of diseases, disorders, and conditions. 128.-. (canceled)29. An isolated binding fusion protein comprising a first targeting moiety with specific binding affinity to a first target and a first extended recombinant polypeptide (XTEN) wherein the XTEN comprises an amino acid sequence which has at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NOS: 41 , 47 , 54 and 59 wherein the XTEN comprises a motif selected from the group consisting of SEQ ID NO: 6 , SEQ ID NO: 7 , SEQ ID NO: 8 , and SEQ ID NO: 9.30. The isolated binding fusion protein of claim 29 , wherein the first targeting moiety binds to a target selected from the group consisting of ANPEP claim 29 , A4 integrin claim 29 , CD19 claim 29 , CD20 claim 29 , CD3 claim 29 , CD3E claim 29 , CD3G claim 29 , CD3Z claim 29 , CD22 claim 29 , CD33 claim 29 , CD74 claim 29 , cMET claim 29 , CTLA4 claim 29 , EGFR claim 29 , EpCAM claim 29 , HER2 claim 29 , ITGAV claim 29 , ITGB3 claim 29 , MUC1 claim 29 , TNFSF8 claim 29 , STEAP1 claim 29 , STEAP2 claim 29 , VEGF claim 29 , mesothilin claim 29 , and carcinoembryonic antigen.31. The isolated binding fusion protein of claim 29 , wherein the first targeting moiety binds to the target with a dissociation constant of less than 10M.32. The isolated binding fusion protein of claim 30 , wherein the first target is CD3.33. The isolated binding fusion protein of that is configured according to formula I:{'br': None, 'sub': x', 'y, '(XTEN)-TM-(XTEN)\u2003\u2003I'}wherein x is either 0 or 1; y is either 0 or 1, wherein x+y≧1, and wherein TM is a targeting moiety ...

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21-01-2016 дата публикации

Process for the Production of Cyclosporin-A Using the Fungus Tolypocladium Sp. Strain NRRL No.: 18950

Номер: US20160017003A1
Принадлежит: Indian Council of Medical Research

Provided herein is a process for the production of Cyclosporin-A (Cyc-A) including the steps of inoculating a nutrient medium with the fungus Tolypocladium sp., strain NRRL No. 18950 followed by cultivation under static conditions to obtain a fermented medium with the fungal biomass, and harvesting the biomass and subjecting the harvested biomass to extraction followed by purification to obtain pure Cyc-A. The nutrient medium includes glucose, glycerol, casein acid hydro lysate, malt extract, peptone, and L-valine.

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18-01-2018 дата публикации

METHOD FOR OBTAINING HIGH-YIELD, STABLE EXPRESSION CELL CLONES AND ANTIBODY MOLECULES OBTAINED THEREBY

Номер: US20180016353A1
Принадлежит:

Provided is a method for obtaining high-yield, stable expression cell clones from myeloma cell lines in a protein-free culture medium. The method is used for industrial production of a recombinant antibody, and includes three stage: (1) adapting to a protein-free culture medium, statically culturing cells at a low density, and gradually reducing a fat-rich supplement to a chemical culture medium; (2) adapting to a protein-free culture medium; culturing cells at a high density, and using a perfusion fermentation system in a laboratory scale; and (3) screening high-yield, stable expression cell clones from the cells after fermentation ends. The cell clone may be used to produce a humanized anti-NeuGcGM3 14F7 recombinant antibody. 1. A method for obtaining stable high producer cell clones from myeloma cell lines in protein-free medium producing recombinant antibodies for industrial purposes that comprise three stages:I. Adaptation to protein-free medium by a stepwise reduction of a lipid-enriched supplement to chemically defined medium in low density stationary cell culture.II. Adaptation to protein-free medium in high density cell culture using perfusion fermentation system at lab scale.III. Selection of stable high producer cell clones from cells at the end of fermentation.2. The method of wherein the first stage comprises the following steps:I. Adaptation to grow in PFHMII cell culture medium+3.5 g/L Cell Boost. Serial passage is performed until Xv reach a constant value.II. Adaptation to grow in PFHMII cell culture medium+1 g/L Cell Boost. Serial passage is performed until Xv reach a constant value.III. Adaptation to grow in PFHMII cell culture medium. Cells are allowed to grow in protein-free medium without any supplement for 60 days.3. The method of wherein the second stage comprises the following steps:{'sup': '6', 'I. Adaptation to grow in PFHMII cell culture medium at high cell density (5-10×10cells/ml) using perfusion fermentation system in 5 L bioreactor for ...

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18-01-2018 дата публикации

METHOD, DEVICE AND KIT FOR MASS CULTIVATION OF CELLS USING POLYIMIDE POROUS MEMBRANE

Номер: US20180016547A1
Принадлежит: UBE INDUSTRIES, LTD.

The present invention pertains to a method for the mass cultivation of cells, and a cell cultivation device and kit. The present invention further pertains to a continuous cell cultivation method and a continuous cell cultivation device in which a carrier is used. 1. A mass cell culturing method including:(1) applying cells to a porous polyimide film, and(2) applying the porous polyimide film to which the cells have been applied, to a cell culture medium and performing culturing.2. The method according to claim 1 , wherein two or more porous polyimide films layered either above and below or left and right are used in the cell culture medium.3. The method according to or claim 1 , wherein the porous polyimide films are:i) folded,ii) wound into a roll,iii) connected as sheets or fragments by a filamentous structure, oriv) bound into a rope,to be suspended or fixed in the cell culture medium in the cell culturing vessel.4. The method according to any one of to claim 1 , wherein in the culturing of step (2) claim 1 , all or some of the porous polyimide films are not in contact with the liquid phase of the cell culture medium.5. The method according to any one of to claim 1 , wherein in the culturing of step (2) claim 1 , the total volume of the cell culture medium in the cell culturing vessel is 10 claim 1 ,000 times or less of the total sum of the porous polyimide film volume including the cell survival zone.6. The method according to any one of to claim 1 , wherein in the culturing of step (2) claim 1 , the total volume of the cell culture medium in the cell culturing vessel is 100 times or less of the total sum of the porous polyimide film volume including the cell survival zone.7. The method according to any one of to claim 1 , wherein the culturing in step (2) is carried out in a system in which a cell culture medium is continuously or intermittently supplied to a cell culturing vessel from cell culture medium supply means installed outside of the cell culturing ...

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18-01-2018 дата публикации

RECOMBINANT PHE-FREE PROTEINS FOR USE IN THE TREATMENT OF PHENYLKETONURIA

Номер: US20180016613A1
Автор: Kämpe Olof, Li Qingshan
Принадлежит: NEXTTOBE AB

The present invention relates to a method for preparing a recombinant Phe-free or Phe-low protein, the method comprising using or as an expression system and/or a recombinant host cell into which a nucleotide encoding a recombinant Phe-free or Phe-low protein has been inserted into the genome. 143-. (canceled)44B. licheniformisB. lichenformis. A method for preparing a recombinant Phe-free or Phe-low protein , the method comprising using a expression system and/or a recombinant host cell into which a nucleotide encoding a recombinant Phe-free or Phe-low protein has been inserted into the genome.45. A method according to claim 44 , wherein the expression system does not contain any antibiotics genes or any spore-formation genes.46B. lichenformis. A method according to claim 44 , wherein the expression system and/or the recombinant host cell is CICC10266.47. A method according to claim 44 , wherein the Phe-free or Phe-low protein is selected from sequences having at least 85% sequence identity with SEQ ID NO: 12.48. A method according to claim 44 , wherein the Phe-free or Phe-low protein is selected from sequences having at least 95% sequence identity with SEQ ID NO: 12.49. A method according to claim 44 , wherein the Phe-free or Phe-low protein is selected from sequences having at least 98% sequence identity with SEQ ID NO: 12.50. A method according to claim 47 , wherein 60% or more of Phe in said sequence has been replaced with one or more LNAA's selected from Tyr claim 47 , Trp claim 47 , Thr claim 47 , Ile claim 47 , Leu claim 47 , Val claim 47 , Met or His.51. A method according to claim 50 , wherein 80% or more of Phe in said sequence has been replaced with one or more LNAA's selected from Tyr claim 50 , Trp claim 50 , Thr claim 50 , Ile claim 50 , Leu claim 50 , Val claim 50 , Met or His.52. A method according to claim 50 , wherein 90% or more of Phe in said sequence has been replaced with one or more LNAA's selected from Tyr claim 50 , Try claim 50 , Thr claim ...

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17-01-2019 дата публикации

MICROBIAL ENGINEERING FOR THE PRODUCTION OF CHEMICAL AND PHARMACEUTICAL PRODUCTS FROM THE ISOPRENOID PATHWAY

Номер: US20190017077A1
Принадлежит:

The invention relates to the production of one or more terpenoids through microbial engineering, and relates to the manufacture of products comprising terpenoids. 1207.-. (canceled)208. A method for making a product containing limonene , or derivative thereof , the method comprising:{'i': Escherichia coli', 'E. coli, 'providing an () that produces isopentyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) through an upstream methylerythritol (MEP) pathway and converts the IPP and DMAPP to limonene, or derivative thereof, through a recombinantly expressed downstream terpenoid synthesis pathway comprising geranyl pyrophosphate synthase and limonene synthase;'}{'i': 'E. coli', 'culturing the to produce the limonene, or derivative thereof, wherein the accumulation of indole in the culture is controlled to below 100 mg/L to thereby increase production of limonene, or derivative thereof; and'}incorporating the limonene, or derivative thereof, into a product.209. The method of claim 208 , wherein the product is a food product claim 208 , food additive claim 208 , beverage claim 208 , chewing gum claim 208 , candy claim 208 , or oral care product.210. The method of claim 208 , wherein the product is a fragrance product claim 208 , a cosmetic claim 208 , a cleaning product claim 208 , or a soap.211. The method of claim 208 , wherein the product is an insecticide claim 208 , pesticide claim 208 , or pest control agent.212. The method of claim 208 , wherein accumulation of indole in the culture is controlled by balancing the upstream MEP pathway with the downstream terpenoid synthesis pathway.213. The method of claim 208 , further comprising measuring the amount or concentration of indole continuously or intermittently.214. The method of claim 208 , wherein accumulation of indole in the culture is maintained to below 50 mg/L.215. The method of claim 208 , wherein accumulation of indole in the culture is maintained to below 10 mg/L.216. The method of claim 208 , ...

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17-04-2014 дата публикации

METHODS FOR PRODUCTION AND PURIFICATION OF POLYPEPTIDES

Номер: US20140106399A1
Принадлежит:

The present invention relates to a method for production and purification of polypeptides. In particular, the present invention relates to a fusion protein comprising a solubility-enhancing peptide tag moiety, a self-aggregating peptide moiety and a moiety of target peptide and to a method for production and purification of target peptides through expressing said fusion protein. 1. A fusion protein comprising a solubility-enhancing peptide tag moiety , a self-aggregating peptide moiety and a target polypeptide moiety , wherein said target polypeptide moiety is located between said solubility-enhancing peptide tag moiety and said self-aggregating peptide moiety , and said target polypeptide moiety is attached to said self-aggregating peptide moiety through a first linker comprising a first cleavage site , wherein said fusion proteins , upon expression in a host cell , are capable of forming active aggregates through said self-aggregating peptide moiety.2. The fusion protein according to claim 1 , wherein said self-aggregating claim 1 , peptide comprises an amphipathic self-assembling short peptide.3. The fusion protein according to claim 2 , wherein said amphipathic self-assembling short peptide is selected from the group consisting of amphipathic β-sheet short peptides claim 2 , amphipathic α-helix short peptides and surfactant-like short peptides.4. The fusion protein according to claim 3 , wherein said self-aggregating peptide moiety comprises one said amphipathic β-sheet short peptide.5. The fusion protein according to claim 3 , wherein said self-aggregating peptide moiety comprises a tandem repeat of two or more of said amphipathic β-sheet short peptides.6. The fusion protein according to claim 4 , wherein said amphipathic β-sheet short peptide is 4-30 amino acid residues in length.7. The fusion protein according to claim 4 , wherein 40%-80% of the amino acid residues in said amphipathic β-sheet short peptide are hydrophobic amino acids.8. The fusion protein ...

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25-01-2018 дата публикации

ANTIBODIES TO IL-6 AND USE THEREOF

Номер: US20180022801A1
Принадлежит:

The present invention is directed to antibodies and fragments thereof and humanized versions thereof having binding specificity for IL-6. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V, Vand CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-IL-6 antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-IL-6 antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-IL-6 antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with IL-6. These antibodies may bind at least one of soluble IL-6, cell surface expressed IL-6, IL-6/IL-6R and/or prevent the association of IL-6 and IL-6R, the association of IL-6/IL-6R and gp130 and or the formation of IL-6/IL-6R/gp130 multimers and thereby inhibit a biological effect associated with any of the foregoing. 1137-. (canceled)138. An isolated host cell which expresses an anti-human IL-6 antibody or antigen-binding fragment which comprises a variable light (V) region comprising complementarity region CDR1 , CDR2 and CDR3 polypeptides , respectively having the sequences of SEQ ID NO:4 , 5 and 6 , and a variable heavy (V) region comprising complementarity region CDR1 , CDR2 and CDR3 polypeptides , respectively having the sequences of SEQ ID NO:7 , 8 or 120 and 9.139. The host cell of claim 138 , wherein the Vregion comprises CDR1 claim 138 , CDR2 and CDR3 polypeptides claim 138 , respectively having the sequences of SEQ ID NO:4 claim 138 , 5 and 6 claim 138 , and the Vregion comprises CDR1 claim 138 , CDR2 and CDR3 polypeptides claim 138 , respectively having the sequences of SEQ ID NO:7 claim 138 , 120 and 9.140. The host cell of claim 138 , wherein ...

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22-01-2015 дата публикации

FISH PROTEIN HYDROLYSATE HAVING A SATIETOGENIC ACTIVITY, NUTRACEUTICAL AND PHARMACOLOGICAL COMPOSITIONS COMPRISING SUCH A HYDROLYSATE AND METHOD FOR OBTAINING SAME

Номер: US20150025001A1
Принадлежит:

The present invention relates to a fish protein hydrolysate containing molecules capable of exerting a satietogenic activity and of regulating food intake in humans or animals. More specifically, the protein hydrolysate according to the invention enables stimulation of the secretion of endogenous cholescystokinins (CCKs) and of endogenous glucagon-like peptide 1 (GLP1) molecules by intestinal cells and the supply of exogenous CCKs. The fish protein hydrolysate according to the invention is obtained by enzymatic hydrolysis of at least one protein source selected from the group composed of the pelagic fish species and spp., the demersal fish species and , and the species of fish belonging to the order Siluriformes, said enzymatic hydrolysis being carried out by means of a mixture of enzymes comprising endopeptidases derived from and from , or derived from , from and from 117-. (canceled)18Bacillus amyloliquefaciensBacillus licheniformisMicromesistius poutassou. A method to exert a satietogenic effect and to regulate food intake comprising , administrating to a human or an animal a pharmaceutical or nutritional composition comprising a and endopeptidases enzyme mixture treated fish protein hydrolysate.19. The method of claim 18 , wherein the fish protein hydrolysate is obtained by a process claim 18 , comprising:{'i': 'Micromesistius poutassou', 'grinding of as protein source,'}{'i': Bacillus amyloliquefaciens', 'Bacillus licheniformis, 'enzymatic hydrolyzing said protein source at a temperature of between 40° and 63° C., at a pH situated between 6 and 9, for 1 to 5 hours, after the addition of a mixture of enzymes comprising endopeptidases derived from and , in a ratio of enzyme to protein source of between 0.01 and 2%, so as to obtain a reaction mixture,'}stopping said enzymatic hydrolysis by inactivation of the said enzymes after raising the temperature of the said reaction mixture to a level not below 70° C., for 8 to 20 minutes, andseparating the protein ...

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25-01-2018 дата публикации

Method For Producing A Recombinant Protein Of Interest

Номер: US20180023067A1
Принадлежит: SANDOZ AG

Disclosed is a method for producing a recombinant protein of interest, characterised in by the following steps: (a) providing a fusion protein comprising an Nautoprotease moiety and a protein of interest moiety in inclusion bodies, (b) solubilising the fusion protein in the inclusion bodies by subjecting the inclusion bodies to a solubilisation buffer containing a detergent and wherein the solubilisation buffer contains no chaotropes or chaotropes in a concentration of less than 1.5 M urea (c) refolding the solubilised fusion protein and (d) allowing the fusion protein to be cleaved by the Nautoprotease moiety under kosmotropic conditions, wherein the recombinant protein of interest is cleaved from the fusion protein, and (e) recovering the protein of interest. 1. Method for producing a recombinant protein of interest , characterised in by the following steps:{'sup': 'pro', '(a) providing a fusion protein comprising an Nautoprotease moiety and a protein of interest moiety in inclusion bodies,'}(b) solubilising the fusion protein in the inclusion bodies by subjecting the inclusion bodies to a solubilisation buffer containing a detergent and wherein the solubilisation buffer contains no chaotropes or chaotropes in a concentration of less than 1.5 M urea(c) refolding the solubilised fusion protein and{'sup': 'pro', '(d) allowing the fusion protein to be cleaved by the Nautoprotease moiety under kosmotropic conditions, wherein the recombinant protein of interest is cleaved from the fusion protein, and'}(e) recovering the protein of interest.2. Method according to claim 1 , characterized in that the inclusion bodies were generated in a recombinant production system.3. Method according to claim 1 , characterized in that the detergent is contained in the solubilisation buffer in a concentration of 0.2 to 15% (w/v).4. Method according to claim 1 , characterized in that steps (c) and/or (d) are performed at kosmotropic conditions that correspond to a urea concentration of 0 ...

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10-02-2022 дата публикации

NOVEL PEPTIDE FOR ENHANCING EXPRESSION EFFICIENCY OF TARGET PROTEIN, AND FUSION PROTEIN COMPRISING SAME

Номер: US20220042063A1
Принадлежит:

The present invention relates to a novel peptide or a partial sequence thereof for enhancing expression efficiency of a target protein, and a fusion protein comprising the same. The novel peptide according to the present invention can enhance expression efficiency of a target protein, and furthermore, the peptide can also be applied to a solubility-enhancing fusion protein in order to enhance solubility of the target protein, so that solubility as well as expression efficiency of the target protein is enhanced, which allows such a peptide to be usefully used for production of a recombinant target protein. 1. A peptide for enhancing expression efficiency of a target protein , the peptide linked to the N-terminus of the target protein and comprising:the amino acid sequence represented by SEQ ID NO: 1 or a partial sequence thereof.2. The peptide according to claim 1 ,wherein the amino acid sequence of SEQ ID NO: 1 is derived from urate oxidase.3. The peptide according to claim 1 ,wherein the peptide contains the amino acid sequence represented by SEQ ID NO: 2.4. The peptide according to claim 1 ,wherein the target protein is at least one selected from the group consisting of antigens, antibodies, cell receptors, enzymes, structural proteins, serum, and cellular proteins.5. A polynucleotide claim 1 , encoding the peptide according to .6. The polynucleotide according to claim 5 ,wherein the polynucleotide is a sequence represented by SEQ ID NO: 3 or SEQ ID NO: 4.7. An expression vector claim 5 , comprising:a polynucleotide encoding a target protein; and{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a polynucleotide encoding the peptide according to , linked to the 5′-end of the polynucleotide encoding the target protein.'}8. The expression vector according to claim 7 ,wherein the target protein is norovirus-derived VP1 protein.9. A host cell claim 7 , transformed with the expression vector according to .10. The host cell according to claim 9 ,{'i': 'E. coli.', ' ...

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10-02-2022 дата публикации

CARBON-SOURCE REGULATED PROTEIN PRODUCTION IN A RECOMBINANT HOST CELL

Номер: US20220042064A1
Принадлежит:

A recombinant host cell comprising an endogenous gene encoding a FLO8 protein comprising the amino acid sequence identified as SEQ ID NO:1 or a homologue thereof, which host cell is engineered by one or more genetic modifications to reduce expression of said gene compared to the host cell prior to said one or more genetic modifications, and which host cell comprises a heterologous expression cassette comprising a gene of interest (GO!) under the control of an expression cassette promoter (ECP) which ECP is repressible by a non-methanol carbon source, and a method of producing a protein of interest using said recombinant host cell. 1. A recombinant host cell comprising an endogenous gene encoding a FLO8 protein comprising the amino acid sequence identified as SEQ ID NO:1 or a homologue thereof , which host cell is engineered by one or more genetic modifications to reduce expression of said gene compared to the host cell prior to said one or more genetic modifications , and which host cell comprises a heterologous expression cassette comprising a gene of interest (GOI) under the control of an expression cassette promoter (ECP) which ECP is repressible by a non-methanol carbon source and comprises:(i) any one of SEQ ID NO:10-16, or(ii) any one of SEQ ID NO:41-45, or(iii) at least 85% sequence identity to a region of at least 300 nt including the 3′end of any one of SEQ ID NO:10-16, or SEQ ID NO:41-45.2. The host cell of claim 1 , wherein said endogenous polynucleotide is a gene encoding said FLO8 protein claim 1 , wherein said one or more genetic modifications comprises a disruption claim 1 , substitution claim 1 , deletion or knockout of (i) one or more endogenous polynucleotides claim 1 , or a part thereof claim 1 , or (ii) an expression control sequence claim 1 , preferably wherein said expression control sequence is selected from the group consisting of a promoter claim 1 , a ribosomal binding site claim 1 , transcriptional or translational start and stop sequences ...

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23-01-2020 дата публикации

EXPRESSION AND LARGE-SCALE PRODUCTION OF PEPTIDES

Номер: US20200024321A1
Принадлежит: Lupin Limited

The invention provides a method for the large-scale preparation of small peptides using recombinant DNA technology. Overexpression of small peptides, such as liraglutide precursor, as concatemers, improves the overall efficiency of the process due to increased yields per batch of the biologically active peptide. Digestion of these concatemers by combinations of specific enzymes yields the desired peptide monomer in large quantities. More particularly, the invention relates to the production of recombinant peptide precursor of liraglutide 1. A concatemeric DNA construct for producing a peptide of SEQ ID 1 , wherein the concatemeric DNA construct comprises:a. DNA construct encoding a peptide of SEQ ID 1, codon optimized for expression in a suitable host{'sup': '−', 'sub': 1', '2, 'claim-text': {'sub': 1', '2, 'wherein Xis Lys or Arg and Xis Lys or Arg;'}, 'b. wherein each unit of (a) is linked at its 3 end to a monomeric or polymeric codon optimized spacer DNA sequence to encode for monomeric or polymeric units of the amino acids X—X,'}c. obtaining concatemeric DNA construct for cloning into a suitable host capable of being expressed as multimers of SEQ ID 1; andd. obtaining multimers of SEQ ID 1, and treating with a combination of at least two proteases to obtain monomeric units of SEQ ID 1.2. The concatemeric DNA construct of claim 1 , wherein the concatemer comprises of at least about 6 monomeric units.3. The concatemeric DNA construct of claim 1 , wherein the DNA construct is at least about 500 bps.4. The concatemeric DNA construct of claim 1 , wherein the DNA construct is expressed in a prokaryotic or eukaryotic host.5. A multimeric peptide of SEQ ID 1 claim 1 , obtainable from the DNA construct of .6. A monomeric peptide of SEQ ID 1 claim 1 , obtainable from the DNA construct of .7. A process for producing a peptide of SEQ ID 1 claim 1 , the process comprising:a. obtaining a codon optimized concatemeric DNA construct encoding for multimers of peptide of SEQ ID 1 ...

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23-01-2020 дата публикации

INCORPORATION OF UNNATURAL NUCLEOTIDES AND METHODS THEREOF

Номер: US20200024597A1
Принадлежит:

Disclosed herein are methods, compositions and kits for the synthesis of proteins which comprises unnatural amino acids that utilize a mutant tRNA. 2. The RNA composition of claim 1 , wherein the codon-anticodon pair read from a 5′ to 3′ direction comprises UUX-XAA claim 1 , UGX-XCA claim 1 , CGX-YCG claim 1 , CGX-XCG claim 1 , AGX-YCU claim 1 , AGX-XCU claim 1 , GAX-YUC claim 1 , GAX-XUC claim 1 , CAX-YUG claim 1 , CAX-XUG claim 1 , GXU-AYC claim 1 , CXU-AYG claim 1 , GXG-CYC claim 1 , AXG-CYU claim 1 , GXC-GYC claim 1 , AXC-GYU claim 1 , or GXA-UYC.3. The RNA composition of claim 2 , wherein the codon-anticodon pair read from a 5′ to 3′ direction comprises CGX-YCG claim 2 , CGX-XCG claim 2 , AGX-XCU claim 2 , GAX-XUC claim 2 , GXU-AYC claim 2 , CXU-AYG claim 2 , GXC-GYC claim 2 , AXC-GYU claim 2 , or GXA-UYC4. The RNA composition of claim 3 , wherein the codon-anticodon pair read from a 5′ to 3′ direction comprises CGX-XCG claim 3 , AGX-XCU claim 3 , GAX-XUC claim 3 , GXU-AYC claim 3 , GXC-GYC claim 3 , or AXC-GYU.5. The RNA composition of claim 4 , wherein the codon-anticodon pair read from a 5′ to 3′ direction comprises GXU-AYC claim 4 , GXC-GYC claim 4 , or AXC-GYU.6. The RNA composition of claim 5 , wherein the codon-anticodon pair read from a 5′ to 3′ direction is GXU-AYC.7. The RNA composition of claim 5 , wherein the codon-anticodon pair read from a 5′ to 3′ direction is GXC-GYC.8. The RNA composition of claim 5 , wherein the codon-anticodon pair read from a 5′ to 3′ direction is AXC-GYU. This application is a continuation of U.S. application Ser. No. 15/543,217, filed on Jul. 12, 2017, which is the U.S. national phase entry of International Application No. PCT/US2016/013095, filed on Jan. 12, 2016, which claims the benefit of U.S. Provisional Application No. 62/102,546, filed on Jan. 12, 2015, each of which are herein incorporated by reference in their entireties.Oligonucleotides and their applications have revolutionized biotechnology. However, the ...

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23-01-2020 дата публикации

COMPOSITIONS AND METHODS OF BIOSYNTHESIZING CAROTENOIDS AND THEIR DERIVATIVES

Номер: US20200024607A1
Автор: Wang Yechun
Принадлежит:

The present invention relates to compositions and methods of producing carotenoids and carotenoid derivatives. 1. A recombinant microorganism comprising at least one nucleic acid construct comprising:a) a nucleic acid sequence encoding a lycopene β-cyclase enzyme; andb) a nucleic acid sequence encoding a carotenoid cleavage dioxygenase enzyme;wherein the nucleic acid sequences are operably linked to one or more expression control sequence, and wherein the microorganism further comprises lycopene.2Mucor circinelloidesPhycomyces blakesleeanusErwinia herbicola.. The microorganism of claim 1 , wherein the lycopene β-cyclase is selected from a lycopene cyclase enzyme of a bifunctional lycopene cyclase/phytoene synthase encoded by carRP of claim 1 , lycopene cyclase/phytoene synthase encoded by carRA of claim 1 , and lycopene cyclase encoded by crtY from3Daucus carota.. The microorganism of claim 1 , wherein the carotenoid cleavage dioxygenase enzyme is encoded by CCD1 from4. The microorganism of claim 1 , wherein the microorganism is selected from a microorganism genetically engineered to inhibit the expression of lycopene α-cyclase and a microorganism naturally not capable of expressing lycopene α-cyclase.5Yarrowia lipolytica, Saccharomyces cerevisiaeE. coli.. The microorganism of claim 1 , wherein the microorganism is selected from claim 1 , and6. The microorganism of claim 1 , comprising β-ionone.7. The recombinant microorganism of claim 1 , wherein the nucleic acid expression construct comprises a nucleic acid sequence encoding a carotenoid cleavage dioxygenase enzyme with at least 80% identity to an amino acid sequence of SEQ ID NO: 88.8. The recombinant microorganism of claim 1 , wherein the nucleic acid expression construct comprises a nucleic acid sequence encoding a lycopene β-cyclase enzyme with at least 80% identity to an amino acid sequence encoded by a nucleic acid sequence selected from SEQ ID NO: 64 and SEQ ID NO: 67.9. A recombinant microorganism ...

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02-02-2017 дата публикации

METHOD FOR PRODUCING ASTAXANTHIN BY FERMENTATION

Номер: US20170029862A1
Принадлежит: JX NIPPON OIL & ENERGY CORPORATION

An object of the present invention is to provide a method for microbioiogically producing astaxanthin of high concentration at low cost while suppressing production of canthaxanthin. Specifically, the present invention relates to a method for producing carotenoids including astaxanthin comprising culturing a bacterium that concurrently produces astaxanthin and canthaxanthin in a medium containing biotin, wherein a ratio of concentration of produced canthaxanthin to concentration of produced astaxanthin in a culture product after the end of culture in the medium is lower than that in a culture product alter the end of culture in a biotin-free medium. 1. A method for producing carotenoids comprising:{'i': 'Paracoccus', '1) culturing a bacterial strain in a culture medium containing biotin at a concentration of 0.02 mg/L to 50 mg/L by addition of biotin to the culture medium, wherein the bacterial strain is a member of the genus and concurrently produces astaxanthin and canthaxanthin;'}2) controlling the concentration of dissolved oxygen in the culture medium at 2 ppm or more during the intermediate phase of culture, thereby producing a carotenoid-containing culture broth in which the ratio or the concentration or canthaxanthin to the concentration of astaxanthin in the culture broth at the end of the culture period is 8% by mass or loss; and3) collecting the carotenoids from the carotenoid-containing culture broth.2. The method of claim further comprising increasing the concentration of dissolved oxygen in the culture medium in a stepwise or continuous manner during the intermediate phase of culture , wherein the concentration of dissolved oxygen in the culture medium at the beginning of the intermediate phase of culture before increasing the concentration is 2 ppm to 3.5 ppm.3. The method of claim 1 , wherein the concentration of gluconic acid in the carotenoid-containing culture broth at the end of the culture period is 30 g/L or less.4. The method of claim 1 , ...

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04-02-2016 дата публикации

METHOD FOR PRODUCING SOLUBLE RECOMBINANT INTERFERON PROTEIN WITHOUT DENATURING

Номер: US20160032345A1
Принадлежит:

The present invention relates to the field of recombinant protein production in bacterial hosts. It further relates to extraction of soluble, active recombinant protein from an insoluble fraction without the use of denaturation and without the need for a refolding step. In particular, the present invention relates to a production process for obtaining high levels a soluble recombinant Type 1 interferon protein from a bacterial host. 1. A method for producing a recombinant Type 1 interferon protein , said method comprising:{'i': Pseudomonas', 'E. coli, 'expressing the recombinant interferon protein by culturing a or host cell containing an expression construct comprising a coding sequence that has been optimized for expression in the host cell;'}lysing the host cell;obtaining an insoluble fraction and a soluble fraction from the lysis step;extracting the insoluble fraction by subjecting it to non-denaturing extraction conditions; andobtaining an extract pellet and an extract supernatant from the insoluble fraction;wherein the recombinant protein in the extract supernatant is present in soluble form, active form, or a combination thereof, without being further subjected to a renaturing or refolding step.2. The method of claim 1 , wherein the non-denaturing extraction conditions comprise the presence of a mild detergent.3. The method of claim 2 , wherein the mild detergent is a Zwitterionic detergent.4. The method of claim 3 , wherein the Zwitterionic detergent is Zwittergent 3-08 claim 3 , Zwittergent 3-10 claim 3 , Zwittergent 3-12 claim 3 , or Zwittergent 3-14.5. The method of claim 4 , wherein the non-denaturing extraction conditions comprise about 0.5% to about 2% Zwittergent 3-14.6. The method of claim 2 , wherein the non-denaturing extraction conditions further comprise a chaotropic agent and a cosmotropic salt.7. The method of claim 6 , wherein the chaotropic agent is urea or guanidinium hydrochloride claim 6 , and wherein the cosmotropic salt is NaCl claim 6 , ...

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04-02-2016 дата публикации

Sortase-mediated protein purification and ligation

Номер: US20160032346A1

The invention relates to a sortase-mediated protein purification and ligation. Specifically, the invention relates to a technique that links protein expression/purification with conjugation to therapeutic agents, imaging agents, or linkers. 1. A conjugation method comprising:cloning a coding sequence of a ligand in series with a coding sequence of sortase recognition, a coding sequence of sortase A, and a coding sequence of an affinity tag;expressing and purifying the protein; andadding calcium and a peptide or protein with an N-terminal glycine,wherein the addition of said calcium and said peptide or protein with an N-terminal glycine allows the sortase to catalyze ligand release and conjugation of the released ligand to said peptide or protein with an N-terminal glycine.2. The method of claim 1 , wherein said sortase recognition sequence comprises LPXTG.3. The method of claim 1 , wherein said affinity tag is a histidine tag.4. The method of claim 1 , wherein said peptide or protein with an N-terminal glycine comprises a functional group.5. The method of claim 1 , wherein said peptide or protein with an N-terminal glycine is linked to a drug molecule claim 1 , an imaging agent claim 1 , a click chemistry group claim 1 , an alkyne claim 1 , an azide claim 1 , a hapten claim 1 , a biotin claim 1 , a protein claim 1 , a small molecule claim 1 , or a nanoparticle.6. The method of claim 1 , said peptide or protein with an N-terminal glycine comprises a plurality of N-terminal glycines.7. A method for purifying a protein claim 1 , the method comprising: the conjugation method of .8. A method for purifying a protein claim 1 , the method comprising:cloning a coding sequence of a ligand protein in series with a coding sequence of sortase recognition, a coding sequence of sortase A, and a coding sequence of an affinity tag;expressing and purifying the protein; andadding calcium and a peptide or protein with an N-terminal glycine,wherein the addition of said calcium and said ...

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01-02-2018 дата публикации

GENETICALLY MODIFIED HOST CELLS AND USE OF SAME FOR PRODUCING ISOPRENOID COMPOUNDS

Номер: US20180030481A1
Принадлежит:

The present invention provides genetically modified eukaryotic host cells that produce isoprenoid precursors or isoprenoid compounds. A subject genetically modified host cell comprises increased activity levels of one or more of mevalonate pathway enzymes, increased levels of prenyltransferase activity, and decreased levels of squalene synthase activity. Methods are provided for the production of an isoprenoid compound or an isoprenoid precursor in a subject genetically modified eukaryotic host cell. The methods generally involve culturing a subject genetically modified host cell under conditions that promote production of high levels of an isoprenoid or isoprenoid precursor compound. 1. A genetically modified eukaryotic host cell that produces an isoprenoid or an isoprenoid precursor compound via a mevalonate pathway , the genetically modified eukaryotic host cell comprising genetic modifications that provide for:a) an increased level of activity of one or more mevalonate pathway enzymes,b) an increased level of prenyltransferase activity, andc) a decreased level of squalene synthase activitywherein the genetic modifications provide for production of an isoprenoid or an isoprenoid precursor compound at a level that is at least about 50% higher than the level of the isoprenoid or isoprenoid precursor compound in a control cell not comprising the genetic modifications.2. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is farnesyl pyrophosphate synthase.3. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is geranyl pyrophosphate synthase.4. The genetically modified eukaryotic host cell of claim 1 , wherein the prenyltransferase is geranylgeranyl pyrophosphate synthase.5. The genetically modified eukaryotic host cell of claim 1 , wherein the genetically modified eukaryotic host cell is a yeast cell.6Saccharomyces cerevisiae.. The genetically modified host cell of claim 5 , wherein the ...

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31-01-2019 дата публикации

Method for Producing a Protein Hydrolysate

Номер: US20190032102A1
Принадлежит: NOVOZYMES A/S

The present invention relates to a method of producing a protein hydrolysate comprising a step of enzymatic protein hydrolysis performed at high temperature. 1. A method for producing a protein hydrolysate , comprising:a) adding to a composition comprising substrate protein a thermostable endopeptidase;b) performing a first hydrolysis step by incubating the composition of step a) for at least 10 minutes at a temperature of at least 75° C.;c) adding to the composition of step b) a protease preparation having an aminopeptidase activity of at least 200 LAPU/g; andd) performing a second hydrolysis step by incubating the composition of step c) for at least 10 minutes at a temperature which is at least 10° C. lower than the temperature used in step b).2. The method of claim 1 , wherein the thermostable endopeptidase is a nonspecific endopeptidase.3. The method of claim 2 , wherein the nonspecific endopeptidase is characterized in that incubation of 0.5% (w/w) BSA with the endopeptidase for 4 hours at a temperature and pH where the endopeptidase exhibits at least 40% of its maximum activity results in a degree of hydrolysis of at least 10%.4. The method of claim 1 , wherein the thermostable endopeptidase is an endopeptidase claim 1 , which after incubation for 15 minutes at 80° C. and pH 9 has a residual activity of at least 80% relative to its activity after incubation at 37° C.5. The method of claim 1 , wherein the thermostable endopeptidase (i) has at least 60% sequence identity to the polypeptide of SEQ ID NO: 3 claim 1 , (ii) is encoded by a polynucleotide having at least 60% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 claim 1 , or (iii) is a variant of the polypeptide of SEQ ID NO: 3 comprising a substitution claim 1 , deletion claim 1 , and/or insertion at one or more positions.6. The method of claim 1 , wherein the thermostable endopeptidase (i) has at least 60% sequence identity to the polypeptide of SEQ ID NO: 8 claim 1 , (ii) is ...

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30-01-2020 дата публикации

STABILIZED PROTEOLYTICALLY ACTIVATED GROWTH DIFFERENTIATION FACTOR 11

Номер: US20200031895A1
Принадлежит:

Methods of activating GDF11 proteins in vitro as well as formulations of mature GDF11 polypeptides with enhanced solubility at neutral pH are provided. 16.-. (canceled)7. An isolated protein comprising , in order , a first amino acid sequence and a second amino acid sequence linked directly via a peptide linker of 5 to 100 amino acids in length , wherein the first amino acid sequence is 52 to 65 amino acids in length and comprises amino acids 60 to 114 or 71-123 of SEQ ID NO:1 , and the second amino acid sequence comprises an amino acid sequence that is at least 90% identical to amino acids 299-407 of SEQ ID NO:1 , wherein the protein when activated by dimerization and/or by dimerization and proteolytic cleavage induces SMAD 2/3 phosphorylation in a Kinase Induced Receptor Activation Assay.833.-. (canceled)34. A nucleic acid sequence encoding the protein of .35. An expression vector comprising the nucleic acid sequence of .36. A host cell comprising the expression vector of .37. A method of producing a protein claim 35 , comprising culturing the host cell of in a culture medium under conditions in which the protein of is produced by the host cell and secreted into the culture medium.38. A method of making an activated GDF11 protein claim 35 , the method comprising:(a) providing a GDF11 protein;(b) subjecting the protein to a disulfide reducing agent to create a first composition;(c) dividing the first composition into a second and a third composition;(d) subjecting the second composition to a cysteine activating agent to create a fourth composition;(e) combining the fourth composition with the third composition to create a fifth composition; and(f) treating the fifth composition with a protease that cleaves at the BMP1 site of the protein, thereby making an optimally activated GDF11 protein.39. The method of claim 38 , wherein the disulfide reducing agent is DTT.40. The method of claim 38 , wherein the cysteine activating agent is aldrithiol.41. The method of claim ...

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30-01-2020 дата публикации

CONSTRUCTS AND CELLS FOR ENHANCED PROTEIN EXPRESSION

Номер: US20200032279A1
Принадлежит: Massachusetts Institute of Technology

Described are expression constructs, cells, and methods of producing proteins in 1. An expression construct comprising an OLE1 promoter operably linked to a nucleic acid encoding a polypeptide comprising a signal sequence and a heterologous protein.2. The expression construct of claim 1 , wherein the signal sequence is identical to the signal sequence of a naturally occurring yeast protein.3. The expression construct of or claim 1 , wherein the signal sequence is the signal sequence of SCW11 claim 1 , MSC1 claim 1 , EXG1 claim 1 , 0841 claim 1 , 1286 claim 1 , BGL2 claim 1 , 2488 claim 1 , 2848 claim 1 , PRY2 claim 1 , 4355 claim 1 , or PIR1.4. The expression construct of any one of to claim 1 , wherein the OLE1 promoter has at least 95% homology with SEQ ID NO: 1 or a fragment thereof.5. The expression construct of claim 4 , wherein the OLE1 promoter has the sequence SEQ ID NO: 1.6. The expression construct of any one of to claim 4 , wherein the expression construct is a plasmid or viral vector.7. The expression construct of claim 6 , wherein the plasmid is an episomal plasmid or an integrative plasmid.8. The expression construct of any one of to claim 6 , wherein the expression construct is linearized.9. The expression construct of any one of to claim 6 , wherein the heterologous protein is selected from the group consisting of enzymes claim 6 , hormones claim 6 , antibodies or antigen binding fragments thereof claim 6 , vaccine components claim 6 , blood factors claim 6 , thrombolytic agents claim 6 , cytokines claim 6 , receptors claim 6 , and fusion proteins.10. A methylotrophic cell expressing a heterologous protein claim 6 , wherein the expression is under the control of an OLE1 promoter.11. The methylotrophic cell of claim 6 , wherein the cell has been transformed by the expression construct of any of -.12. The methylotrophic cell of or claim 6 , wherein the OLE1 promoter is located at the OLE1 claim 6 , AOX1 claim 6 , GAPDH claim 6 , DAS2 claim 6 , or PIF1 ...

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30-01-2020 дата публикации

METHOD FOR PRODUCING USEFUL MATERIAL

Номер: US20200032313A1
Принадлежит: SANYO CHEMICAL INDUSTRIES, LTD.

The present invention provides a method of producing a useful substance, including secreting and producing a useful substance in broth by microorganisms contained in the broth, wherein the microorganisms include microorganisms of at least one genus selected from the group consisting of , and , and the broth contains at least one surfactant (A) selected from the group consisting of an anionic surfactant (A1), a cationic surfactant (A2), and an amphoteric surfactant (A3). 1. A method of producing a useful substance , comprising:secreting and producing a useful substance in broth by microorganisms contained in the broth,{'i': Ogataea, Saccharomyces, Kluyveromyces, Hansenula, Pichia, Yarrowia', 'Candida, 'wherein the microorganisms comprise microorganisms of at least one genus selected from the group consisting of , and , and the broth contains at least one surfactant (A) selected from the group consisting of an anionic surfactant (A1), a cationic surfactant (A2), and an amphoteric surfactant (A3).'}2. The method of producing a useful substance according to claim 1 ,wherein the amount of the surfactant (A) in the broth is 0.0001 to 20 wt % based on the weight of the broth.3. The method of producing a useful substance according to claim 1 ,wherein the useful substance is a protein.4. The method of producing a useful substance according to claim 2 ,wherein the useful substance is a protein. The present invention relates to a method of producing a useful substance.Yeast has been widely used to produce useful substances such as amino acids and proteins. Particularly in recent years, a technique has been known in which proteins are efficiently produced by yeast transformed by introducing a medically and/or industrially useful protein gene thereinto.Use of as a bacterium for production of useful substances is limited to production of proteins without sugar moieties because a glycan synthetase is absent in . In contrast, use of yeast for production of useful substances can ...

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30-01-2020 дата публикации

CO-PRODUCTION OF A SESQUITERPENE AND A CAROTENOID

Номер: US20200032314A1
Принадлежит:

Provided herein are compositions and methods for co-production and recovery of two or more isoprenoids from a single recombinant cell. 1. A method for co-production of two or more isoprenoids , the method comprising:(a) culturing, in a culture medium, a host cell genetically modified with one or more heterologous nucleic acids encoding one or more enzymes in a first biosynthetic pathway to produce a first isoprenoid and with one or more heterologous nucleic acid encoding one or more enzymes in a second biosynthetic pathway to produce a second isoprenoid, which has a molecular weight that is different from the first isoprenoid; and(b) recovering the first isoprenoid; and(c) recovering the second isoprenoid.2. The method of wherein the host cell is not genetically modified to produce a target compound for recovery other than a compound derived from IPP.3. The method for co-production of isoprenoids of claim 1 , wherein the first isoprenoid and second isoprenoid are produced concurrently during a fermentation run from a single inoculum.4. The method for co-production of isoprenoids of claim 1 , wherein the first isoprenoid and second isoprenoid are produced sequentially from a single inoculum comprising the host cell claim 1 , or sequentially using a genetic switch.5. (canceled)6. The method for co-production of isoprenoids of claim 1 , wherein the culturing and recovering comprise:(a) culturing the single inoculum comprising the host cell to build a population of host cells;(b) culturing the population of host cells under conditions to produce the first isoprenoid from the population of host cells, wherein the conditions do not activate production of the second isoprenoid;(c) recovering the first isoprenoid from the population;(d) after separating the first isoprenoid, culturing the population or a subpopulation of the host cells under conditions to activate production of the second isoprenoid; and(e) recovering the second isoprenoid.7. The method for co-production of ...

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05-02-2015 дата публикации

PRODUCTION OF ASTAXANTHIN AND DOCOSAHEXAENOIC ACID IN MIXOTROPHIC MODE USING SCHIZOCHYTRIUM

Номер: US20150037838A1
Принадлежит: FERMENTALG

New strains of protists belonging to the genus, allow high-yield production of lipids and carotenoids, in particular of astaxanthin and docosahexaenoic acid (DHA), in mixotrophic mode, and a method for selecting and culturing such strains, using a variable and/or discontinuous light source, in particular a flashing light. 1. Method comprising the following step:{'i': 'Schizochytrium', 'sup': −2', '−1', '−2', '−1, 'a) culture, in mixotrophic mode, of one or more strains of the genus under conditions of illumination that is discontinuous and/or variable over time, the illumination having variations in intensity, the amplitude of which is comprised between 5 μmol·m·sand 1,000 μmol·m·s, these variations taking place between 2 and 3,600 times per hour.'}2. Method according to claim 1 , characterized in that the illumination has variations in intensity claim 1 , the amplitude of which is comprised between 5 μmol·m·sand 400 μmol·m claim 1 , these variations taking place between 2 and 200 times per hour.3. Method according to claim 1 , characterized in that the culture is carried out in the presence of an organic carbon-containing substrate at a concentration from 100 mM to 1.5 M claim 1 , preferably from 300 mM to 1.2 M claim 1 , more preferentially from 500 mM to 1 M claim 1 , and even more preferentially from 600 mM to 900 mM claim 1 , the organic carbon-containing substrate being selected from saccharose claim 1 , glycerol claim 1 , glucose and cellulose derivatives and a mixture of these molecules.4. Method according to claim 3 , characterized in that said organic carbon-containing substrate present in the culture medium comprises at least 300 mM of glucose and/or glycerol.5. Method according to claim 1 , characterized in that the amplitude of the variations in intensity is comprised between 5 and 1 claim 1 ,000 μmol·m·s claim 1 , preferably between 70 and 300 μmol·m·s claim 1 , and more preferentially between 100 and 200 μmol·m·s.6. Method according to claim 1 , ...

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05-02-2015 дата публикации

PRODUCTION OF LUTEIN IN MIXOTROPHIC MODE BY SCENEDESMUS

Номер: US20150037839A1
Принадлежит: FERMENTALG

Novel strains of microalgae belonging to the genus enable the production of lipids, in particular lutein, in mixotrophic mode, as well as a method for selecting and culturing the strains using a variable and/or discontinuous supply of light, in particular in the form of flashes. 1. Method comprising the following step:{'i': 'Scenedesmus', 'sup': −2', '−1', '−2', '−1, 'a) culture, in mixotrophic mode, of one or more strains of the genus under conditions of illumination that is discontinuous and/or variable over time, the illumination having variations in intensity, the amplitude of which is comprised between 5 and 1,000, 5 μmol·m·sand 400 μmol·m·s, these variations taking place between 2 and 3,600 times per hour.'}2. Method according to claim 1 , characterized in that the illumination has variations in intensity claim 1 , the amplitude of which is comprised between 5 μmol·m·sand 400 μmol·m claim 1 , these variations taking place between 2 and 200 times per hour.3Scenedesmus. Method according to claim 1 , characterized in that the microalga is from the species sp.4. Method according to claim 1 , characterized in that the culture is carried out in the presence of an organic carbon-containing substrate at a concentration from 5 mM to 1 M claim 1 , preferably from 50 mM to 800 mM claim 1 , more preferentially from 70 mM to 600 mM claim 1 , and even more preferentially from 100 mM to 500 mM claim 1 , the organic carbon-containing substrate being selected from starch claim 1 , lactate claim 1 , lactose claim 1 , saccharose claim 1 , acetate claim 1 , glycerol claim 1 , glucose and cellulose derivatives and a mixture of these molecules.5. Method according to claim 4 , characterized in that said organic carbon-containing substrate present in the culture medium comprises at least 5 mM of glucose.6. Method according to claim 5 , characterized in that the amplitude of the variations in intensity is comprised between 5 and 1 claim 5 ,000 claim 5 , preferably between 30 and 400 μ ...

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05-02-2015 дата публикации

Use of Molecular Chaperones for the Enhanced Production of Secreted, Recombinant Proteins in Mammalian Cells

Номер: US20150037841A1
Принадлежит:

The present invention relates to a method for increased production of a secreted, recombinant protein product through the introduction of molecular chaperones in a mammalian host cell. The present invention also relates to a mammalian host cell with enhanced expression of a secreted recombinant protein product by coexpressing at least one chaperone protein. 139-. (canceled)40. A method for producing a secreted recombinant protein product comprising the steps of:culturing a mammalian host cell, said mammalian host cell having genetic material coding for expression of said recombinant protein product and transformed with at least one expression vector comprising DNA encoding a chaperone protein selected from the group consisting of calnexin, calreticulin, Erp57, hsp40, and Hsp70; andrecovering from the culture medium the recombinant protein product so produced and secreted.41. The method according to claim 40 , wherein the genetic material coding for expression of said recombinant protein product is integrated into the host cell DNA.42. The method according to claim 41 , wherein said mammalian host cell is further transformed with an expression vector comprising DNA encoding a glutamine synthetase protein.43. (canceled)44. (canceled)45. The method according to claim 40 , wherein the recombinant protein is Factor VIII protein or fragment thereof.4654-. (canceled)55. A method for enhancing yield of recombinant Factor VIII or fragment thereof in a baby hamster kidney (BHK) cell line claim 40 , wherein genetic material coding for expression of said recombinant Factor VIII or fragment thereof has been previously introduced into a first BHK cell line claim 40 , said method comprising the steps of:inserting at least one chaperone protein expression vector into said first BHK cell line so as to form a modified BHK cell line; andselecting from said modified BHK cell line at least one second cell line exhibiting enhanced yield of the recombinant Factor VIII or fragment thereof ...

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11-02-2016 дата публикации

METHOD FOR STABILISING OXIDATION-SENSITIVE METABOLITES PRODUCED BY MICROALGAE OF THE CHLORELLA GENUS

Номер: US20160040208A1
Автор: MACQUART GABRIEL
Принадлежит:

The invention relates to a method for stabilising a biomass of microalgae containing oxidation-sensitive metabolites selected from the group consisting of carotenoids (lutein, etc.), monounsaturated and polyunsaturated fatty acids (palmitoleic acid, oleic acid, linoleic acid, etc.), chlorophyll pigments (chlorophyll A and B, etc.) and vitamins (vitamin B9 and B12, etc.) taken individually or together, more specifically carotenoids, said method comprising the fermentation of said biomass in heterotrophic conditions. 113-. (canceled)14. A method for stabilizing or for storing oxidation-sensitive metabolites selected from the group consisting of the carotenoids , monounsaturated and polyunsaturated fatty acids , chlorophyll pigments , and vitamins , alone or in combination comprising:fermenting a biomass of microalgae in heterotrophic conditions comprising a culture phase deficient in a nutrient factor; andstoring the dry biomass in which the oxidation-sensitive metabolites are stabilized.15Chlorella.. The method of claim 14 , wherein the microalgae are of the genus16Chlorella sorokiniana.. The method of claim 15 , wherein the microalgae are17. The method of claim 15 , wherein the method does not comprise adding exogenous antioxidant or stabilizer to said dry biomass.18. The method of claim 15 , wherein the deficient nutrient factor is the carbon-containing source.19. The method of claim 18 , wherein the fermenting of said biomass of microalgae in heterotrophic conditions comprises:a first step of fermentation in batch mode,a second step of fermentation in fed-batch mode which, when the carbon-containing source is completely consumed by the microalgae, involves continuous supply of said carbon-containing source at a rate lower than its rate of consumption by the microalgae.20. The method of claim 18 , wherein the deficient nutrient factor is glucose and in that it is supplied to the culture at a rate above 1 g/l/h.21. The method of claim 19 , wherein the deficient ...

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08-02-2018 дата публикации

SYNTHETIC PEPTIDES AND ENZYMATIC FORMATION OF INTRACELLULAR HYDROGELS

Номер: US20180037605A1
Автор: DU Xuewen, Xu Bing, ZHOU JIE
Принадлежит:

The invention relates to a peptide that includes a plurality of amino acid residues and an enzymatically cleavable moiety including taurine or hypotaurine, the enzymatically cleavable-moiety being linked to the peptide via covalent bond, wherein the peptide is capable of self-assembly to form nanofibrils in the presence of an enzyme that hydrolyzes the enzymatically cleavable-moiety. Compositions containing the enzymatically responsive peptide, and the use thereof for forming a nanofibril network internally of cells, for treating a cancerous condition, and imaging cells are also disclosed. 1. A peptide comprising a plurality of amino acid residues and an enzymatically cleavable moiety comprising a taurine or hypotaurine residue , the enzymatically cleavable-moiety being linked to the peptide via covalent bond , wherein the peptide is capable of self-assembly to form nanofibrils in the presence of an enzyme that hydrolyzes the enzymatically cleavable-moiety.2. The peptide according to claim 1 , wherein the amino acids are aromatic amino acids selected from the group consisting of phenylalanine claim 1 , phenylalanine derivatives claim 1 , tyrosine claim 1 , tyrosine derivatives claim 1 , tryptophan claim 1 , and tryptophan derivatives.3. The peptide according to claim 1 , wherein the amino acids are all D-amino acids or all L-amino acids.4. The peptide according to claim 1 , wherein the amino acids are a mixture of L-amino acids and D-amino acids.5. The peptide according to claim 1 , wherein a taurine residue is present.67-. (canceled)8. The peptide according to claim 1 , wherein the covalent bond linking the enzymatically cleavable-moiety to the peptide is a peptide bond.9. The peptide according to claim 1 , wherein the enzymatically cleavable-moiety further comprises an ester claim 1 , a carbonate claim 1 , a thiocarbonate claim 1 , a carbamate claim 1 , a carboxylate claim 1 , a diacyl anhydride claim 1 , or an amide bond.1113-. (canceled)14. The peptide according ...

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08-02-2018 дата публикации

FUNGAL STRAINS AND METHODS OF USE

Номер: US20180037919A1
Принадлежит: DANISCO US INC.

Provided are improved fungal strains and use thereof, wherein the fungal strains are capable of producing an altered level of proteins, enzymes, variants and other substances of interest. 1. An engineered fungal strain , capable of producing an altered level of a protein of interest as compared to a parental strain , wherein the engineered fungal strain comprises a variant histidine kinase gene.2. An engineered fungal strain , capable of producing an altered level of a protein of interest as compared to a parental strain , wherein the engineered fungal strain comprises a mutation that causes altered sensitivity or resistance to external osmotic pressure as compared to the parental strain.3. An engineered fungal strain , capable of producing an altered level of a protein of interest as compared to a parental strain , wherein the engineered fungal strain comprises a mutation that causes altered sensitivity or resistance to a fungicide as compared to the parental strain.49-. (canceled)10. The engineered fungal strain of claim 1 , wherein the parental strain is an Ascomycete fungal strain.1112-. (canceled)13. A transformed fungal strain or a derivative fungal strain thereof capable of producing an altered level of a protein of interest as compared to a parental strain claim 1 , wherein the transformed fungal strain or the derivative fungal strain comprises a variant histidine kinase gene or a mutation that causes altered sensitivity or resistance to external osmotic pressure as compared to the parental strain.1419-. (canceled)20. The fungal strain of claim 13 , wherein the parental strain is an Ascomycete fungal strain.2129-. (canceled)3023. The method of claim claim 13 , wherein the parental strain is an Ascomycete fungal strain.3132-. (canceled)33. A method of producing a protein of interest comprising fermenting the engineered fungal strain of claim 1 , wherein the engineered claim 1 , transformed or derivative fungal strain secretes the protein of interest claim 1 , ...

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08-02-2018 дата публикации

ANTIBODIES CONJUGATABLE BY TRANSGLUTAMINASE AND CONJUGATES MADE THEREFROM

Номер: US20180037921A1
Принадлежит:

An antibody has at a heavy chain thereof a C-terminal extension that includes at least one glutamine that is a substrate for transglutaminase, enabling the transglutaminase-mediated preparation antibody-drug conjugates using such antibody. 1. A full length antibody , wherein at least one heavy chain has a C-terminal extension having an amino acid sequence comprising EEQYASTY (SEQ ID NO:1) , EEQYQSTY (SEQ ID NO:2) , EEQYNSTY (SEQ ID NO:3) , EEQYQS (amino acids 1 through 6 of SEQ ID NO:2) , EEQYQST (amino acids 1 through 7 of SEQ ID NO: 2) , EQYQSTY (amino acids 2 through 8 of SEQ ID NO:2) , QYQS (amino acids 3 through 6 of SEQ ID NO: 6) , or QYQSTY (amino acids 3 through 8 of SEQ ID NO: 2).2. A full length antibody according to claim 1 , wherein the C-terminal extension has an amino acid sequence comprising EEQYASTY (SEQ ID NO:1) claim 1 , EEQYQSTY (SEQ ID NO:2) or EEQYNSTY (SEQ ID NO:3).3. A full length antibody according to claim 1 , wherein the C-terminal extension has an amino acid sequence consisting of EEQYASTY (SEQ ID NO:1).4. A full length antibody according to claim 1 , wherein the C-terminal extension has an amino acid sequence consisting of EEQYQSTY (SEQ ID NO:2).5. A full length antibody according to claim 1 , wherein the C-terminal extension has an amino acid sequence consisting of EEQYNSTY (SEQ ID NO:3).6. A full length antibody according to claim 1 , wherein the C-terminal extension has an amino acid sequence consisting of EEQYQS (amino acids 1 through 6 of SEQ ID NO:2) claim 1 , EEQYQST (amino acids 1 through 7 of SEQ ID NO: 2) claim 1 , EQYQSTY (amino acids 2 through 8 of SEQ ID NO:2) claim 1 , QYQS (amino acids 3 through 6 of SEQ ID NO: 6) claim 1 , or QYQSTY (amino acids 3 through 8 of SEQ ID NO: 2).7. A full length antibody according to claim 1 , which is of the IgG1 isotype.8. A full length antibody according to claim 1 , having a constant region having an amino acid sequence according to SEQ ID NO:4.9. A method of making an antibody-drug ...

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08-02-2018 дата публикации

Recombinase Purification

Номер: US20180037922A1
Автор: Longo Michael
Принадлежит:

The invention is the products and methods associated with purifying overexpressed recombinant recombinases from a host cell line resulting in an un-tagged protein of interest without any additional, non-native amino acids. The invention employs at least one DNA vector that co-expresses a tagged fusion protein and the recomibinase protein with the recombinase protein having an affinity for binding to the the tagged fusion protein. Isolation methods of the recominbase protein include the targeting of the tagged fusion protein. 1. A method of purifying a protein , comprising:a. inserting at least one expression vector into a host cell line, wherein said at least one expression vector comprises at least a first coding sequences and a second coding sequence, and the first coding sequence and the second coding sequence is under the control of at least one promoter, wherein the first coding sequence encodes for a tagged fusion protein comprising a BRCA2 protein motif, and the second coding sequence encodes for a recombinase protein;b. expressing the tagged fusion protein and the recombinase protein from under the control of the at least one promoter in the host cell line; andc. isolating the recombinase protein from the host cell line using a protein purification procedure that comprises of procedures that select for the tagged fusion protein.2. The method of wherein the BRCA2 protein motif comprises a BRC4 F-X-X-A motif.3. The method of claim 1 , wherein the tagged fusion protein comprises at least one protein tag.4. The method of wherein said at least one protein tag comprises a 6×his tag claim 3 , an MBP tag claim 3 , a GST tag claim 3 , a FLAG tag claim 3 , a myc tag claim 3 , or a Strep tag or any combination of protein tags.5. The method of claim 3 , wherein the first coding sequence further comprises a 6×his tag and an MBP tag and the BRC4 protein motif claim 3 , wherein the first coding sequence codes for a protein as set forth in SEQ ID NO: 22.6. The method of ...

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