ФАРМАЦЕВТИЧЕСКИЙ ПРЕПАРАТ И СПОСОБ ЛЕЧЕНИЯ ЗЛОКАЧЕСТВЕННЫХ ОПУХОЛЕЙ У ЧЕЛОВЕКА С ПОМОЩЬЮ АРГИНИНОВОЙ ДЕПРИВАЦИИ

Изобретение относится к биотехнологии. Описана выделенная и модифицированная рекомбинантная аргиназа I человека, имеющая чистоту 80-100%, приводящая к периоду полужизни в плазме in vivo, по меньшей мере, приблизительно 3 дня и к удельной активности, по меньшей мере, 250 МЕ/мг, где указанная аргиназа модифицирована сшиванием с не иммуногенной молекулой. Представлена фармацевтическая композиция для снижения уровня аргинина, содержащая описанную рекомбинантную аргиназу I человека в эффективном количестве. Предложен способ лечения злокачественных опухолей человека, предусматривающий введение описанной рекомбинантной аргиназы I человека или фармацевтической композиции, содержащей описанную аргиназу, внутрь пациента. Изобретение позволяет эффективно снижать уровень аргинина у пациентов с опухолями и может быть использовано для лечения злокачественных опухолей человека. 5 н. и 12 з.п. ф-лы, 47 ил., 3 табл.

СПОСОБ ПОЛУЧЕНИЯ ЩЕЛОЧНЫХ ГИДРОЛИЗАТОВ РАСТИТЕЛЬНЫХ БЕЛКОВ

Группа изобретений относится к пищевой, сельскохозяйственной, косметической и фармацевтической отраслям. Получают суспензию растительных белков, выбранных из группы, включающей гороховые белки, картофельные белки и кукурузные белки, с содержанием сухого вещества от 10 до 15%. Доводят рН смеси при перемешивании до значения от 9,5 до 10,5 в качестве единственного щелочного средства при помощи одного или нескольких гидроксидов щелочных металлов, выбранных из группы, включающей гидроксид натрия и гидроксид калия. Нагревают полученную суспензию при температуре от 70 до 80°С в течение от 4 до 6 ч. Нейтрализуют нагретую суспензию посредством минеральной кислоты, предпочтительно хлористоводородной кислоты. Сушат нейтрализованную суспензию для получения щелочного гидролизата. Щелочной гидролизат растительных белков имеет значение водорастворимости при рН 7,5 от 60 до 100%, предпочтительно от 80 до 98%, эмульгирующую способность от 60 до 90%, предпочтительно от 65 до 85%, среднюю длину пептидной ...

СПОСОБ ПОДАВЛЕНИЯ ВИЧ-ИНФИЦИРОВАННЫХ КЛЕТОК МЛЕКОПИТАЮЩЕГО И БЕЛКОВЫЙ РЕКОМБИНАНТНЫЙ РЕЦЕПТОР ДЛЯ ЕГО ОСУЩЕСТВЛЕНИЯ

Изобретение относится к области генной инженерии. Предложенный способ включает введение млекопитающему эффективного количества терапевтических клеток, экспрессирующих связанный с мембраной белковый рекомбинантный рецептор, включающий в себя внеклеточную часть, которая включает в себя фрагмент CD4, который обладает способностью специфически узнавать и связывать ВИЧ-инфицированную клетку, но который не опосредует ВИЧ-инфицирование. Кроме того, описан белковый рекомбинантный рецептор, который также способен специфично распознавать и связывать ВИЧ-инфицированную клетку, но который не опосредует ВИЧ-инфекцию. Изобретение позволяет управлять клеточным иммунным ответом, направленным против ВИЧ-инфицированной клетки млекопитающего. 2 с. и 9 з.п. ф-лы, 28 ил., 1 табл.

ПИЩЕВОЙ БЕЛКОВЫЙ ИНГРЕДИЕНТ И СПОСОБЫ ЕГО ПОЛУЧЕНИЯ

Группа изобретений относится к вариантам способа получения пищевого белкового ингредиента, а также пищевому белковому ингредиенту, полученному способом. Предложен способ получения пищевого белкового ингредиента, включающий обработку содержащего белок материала восстанавливающим агентом, хаотропным агентом, детергентом или их смесями. Дополнительно может быть осуществлено приведение содержащего белок материала в контакт с водой. Осуществляют нагревание приведенного в контакт содержащего белок материала высоконасыщенным паром для получения белковой смеси до тех пор, пока приведенный в контакт содержащий белок материал не достигнет 85-95°C. Получают раствор протеолитического фермента путем смешивания протеолитического фермента в водном растворе. Приводят белковую смесь в контакт с раствором протеолитического фермента для получения смеси белка с ферментом с дальнейшим уменьшения размера для получения смеси белка меньшего размера с ферментом. Инкубируют смесь белка меньшего размера с ферментом ...

МУТАНТНЫЕ РЕЦЕПТОРЫ И ИХ ИСПОЛЬЗОВАНИЕ В СИСТЕМЕ ЭКСПРЕССИИ ИНДУЦИБЕЛЬНОГО ГЕНА НА ОСНОВЕ ЯДЕРНОГО РЕЦЕПТОРА

Изобретение относится к области биотехнологии или генной инженерии. Описана система модулирования экспрессии рекомбинантного гена, вектор экспрессии и клетка-хозяин. Изобретение расширяет арсенал методов модулирования экспрессии гена. 3 н. и 9 з.п. ф-лы, 14 табл., 10 пр.

МУЛЬТИПЛЕКСНЫЙ АНАЛИЗ СТОПОЧНОГО ТРАНСГЕННОГО БЕЛКА

Изобретение относится к биотехнологии, а именно к способу выявления присутствия двух или более представляющих интерес белков с известными аминокислотными последовательностями в образце растительного происхождения (варианты) и способу сохранения генотипа сорта трансгенного растения. Способы включают расщепление всех белков из образца растительного происхождения специфичной по последовательности протеазой с получением пептидов. Разделяют пептиды в один прием в приборе для жидкостной хроматографии - масс-спектрометрии (ЖХ-МС). Ионизируют пептиды в приборе для ЖХ-МС. Проводят одновременное получение масс-спектральных данных для пептидов в приборе для ЖХ-МС. Сравнивают указанные одновременные масс-спектральные данные для пептидов с ожидаемыми масс-спектральными данными от представляющих интерес белков с известными аминокислотными последовательностями. Осуществляют определение присутствия или отсутствия представляющих интерес белков. Предложенное изобретение позволяет с высокой эффективностью ...

СПОСОБ ВЫДЕЛЕНИЯ РЕКОМБИНАНТНЫХ БЕЛКОВ

Изобретение относится к области биотехнологии и может быть использовано при получении рекомбинантных форм представляющих практический интерес белков. Предложен способ получения рекомбинантного белка через его гибридный предшественник с природным сайтом расщепления энтеропептидазой, который обеспечивает повышение качества (гомогенности) и выхода целевого продукта в условиях, когда гибридный белок обнаруживает дополнительные (скрытые) сайты расщепления энтеропептидазой. Результат достигается путем замены в природном сайте расщепления энтеропептидазой Асп-Асп-Асп-Асп-Лиз аминокислотного остатка лизина (Лиз) аминокислотным остатком аргинина (Арг) и последующего расщепления гибридного предшественника легкой каталитической субъединицей энтеропептидазы человека или быка. 3 табл., 3 ил., 4 пр.

ТРИПЕПТИДИЛПЕПТИДАЗЫ, СПОСОБНЫЕ ВОЗДЕЙСТВОВАТЬ НА СВЯЗИ С УЧАСТИЕМ ПРОЛИНА, И ИХ ПРИМЕНЕНИЯ

Изобретение относится к области биотехнологии. Описан способ получения гидролизата. Заявленный способ включает стадию смешивания белка или его части с эндопротеазой и толерантной к пролину трипептидилпептидазой, обладающей экзопептидазной активностью, где указанная толерантная к пролину трипептидилпептидаза способна к отщеплению трипептидов с N-конца пептидов, имеющих пролин в P1' или аминокислоту, выбранную из аланина, аргинина, аспарагина, аспарагиновой кислоты, цистеина, глутамина, глутаминовой кислоты, глицина, гистидина, изолейцина, лейцина, лизина, метионина, фенилаланина, серина, треонина, триптофана, тирозина, валина или синтетических аминокислот в положении P1'. Изобретение расширяет арсенал средств для получения гидролизата. 14 з.п. ф-лы, 28 ил., 13 табл., 24 пр.

Устройство для гидролиза водных растворов биополимеров

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.

СПОСОБ ПОЛУЧЕНИЯ ШТАММА-ПРОДУЦЕНТА ГИБРИДНОГО БЕЛКА APRO INS ДЛЯ ПОЛУЧЕНИЯ ИНСУЛИНА ЧЕЛОВЕКА

Изобретение относится к биотехнологии. Способ создания штамма E.coli - продуцента гибридного белка AproIns для получения инсулина человека включает конструирование полного структурного гена проинсулина с помощью химико-фармацевтического синтеза, моделирование и расчет оптимальной структуры IgG - связывающего домена белка А, синтез наиболее оптимальной структуры лидерного пептида, связывание его с образованием гибридного белка AproIns, перенос в экспрессирующий вектор и трансформацию клеток E.coli. Способ позволяет создать наиболее оптимальную структуру гибридного белка для получения инсулина человека по простой и безопасной технологии. 2 з.п.ф-лы, 9 ил.

БИОЛОГИЧЕСКИ АКТИВНЫЙ ПЕПТИД, ПОЛУЧЕННЫЙ ИЗ МОЛОЧНОГО БЕЛКА

Изобретение относится к области биотехнологии, а именно к биологически активному пептиду, который обладает антигипертонической активностью. Предложенное изобретение может быть использовано для профилактики гипертонических заболеваний. Биологически активный пептид характеризуется аминокислотной последовательностью LLYQQPVLGPVRGPFPIIV. Данный пептид получают из белка молока ферментативным гидролизом. После чего осуществляют очистку и ультрафильтрацию полученных гидролизатов с использованием мембран с диаметром пор 10 и 15 кД при рН 6,0-6,5. Предложенное изобретение позволяет получать биологически активный пептид, который обладает антигипертонической активностью.

СПОСОБ ПОЛУЧЕНИЯ АНАЛОГОВ ИНСУЛИНА ИЗ ИХ СООТВЕТСТВУЮЩИХ ПРЕДШЕСТВЕННИКОВ (ВАРИАНТЫ)

Изобретение относится к области биотехнологии, конкретно к получению аналогов инсулина из их соответствующих предшественников, и может быть использовано в медицине. Способ получения аналогов инсулина из их предшественников предусматривает проведение одностадийной ферментативной реакции, включающей комбинаторное и одновременное использование оптимальных количеств трипсина и карбоксипептидазы В, которые действуют синергично, направляя реакцию управляемым способом, для того чтобы избежать продукции случайных нежелательных побочных продуктов. Изобретение позволяет сократить количество операционных стадий и получить аналоги инсулина с более высоким выходом и чистотой. 2 н. и 18 з.п. ф-лы, 5 пр.

ПОЛНОСТЬЮ ЧЕЛОВЕЧЕСКИЕ АНТИ-VEGF-АНТИТЕЛА И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

... 1. Изолированное антитело или его антигенсвязывающий фрагмент, которое специфично связывает VEGF, и содержащее одну или несколько из следующих характеристик: ! a) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 6, SEQ ID NO: 7 и SEQ ID NO: 8; ! b) вариабельную область легкой цепи, содержащую SEQ ID NO: 9, SEQ ID NO: 10 и SEQ ID NO: 11; ! c) вариабельную область легкой цепи, содержащую SEQ ID NO: 12, SEQ ID NO: 13 и SEQ ID NO: 14; ! d) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 15; ! e) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 6, SEQ ID NO: 7 и SEQ ID NO: 15; ! f) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 16; ! g) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 6, SEQ ID NO: 7 и SEQ ID NO: 16; ! h) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 17; ! i) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 6, SEQ ID NO: 7 и SEQ ID NO: 17; ! j) вариабельную область тяжелой цепи, содержащую SEQ ID NO: 18; ! k) вариабельную ...

СПОСОБЫ И МАТЕРИАЛЫ ДЛЯ ПОЛУЧЕНИЯ СИМВАСТАТИНА И РОДСТВЕННЫХ СОЕДИНЕНИЙ

... 1. Способ получения симвастатина, включающий стадии: !(1) взаимодействия монаколина J; ацилтиоэфира, который отдает ацильную группу C8-гидроксильной группе монаколина J в присутствии ацилтрансферазы LovD; и ацилтрансферазы LovD; и ! (2) региоселективного ацилирования C8-гидроксильной группы монаколина J ацилтрансферазой LovD, использующей ацильную группу ацилтиоэфира, с получением симвастатина. ! 2. Способ по п.1, где симвастатин получают in vitro в отсутствии выделенного организма. ! 3. Способ по п.1, где монаколин J; ацилтиоэфир и ацилтрансфераза LovD взаимодействуют в среде культивирования в присутствии выделенного организма, который продуцирует ацилтрансферазу LovD, и где дополнительно организм представляет собой Escherichia coli, Aspergillus terreus, Monascus ruber, Monascus purpureus, Monascus pilosus, Monascus vitreus, Monascus pubigerus, Candida cariosilognicola, Aspergillus oryzea, Doratomyces stemonitis, Paecilomyces virioti, Penicillum citrinum, Penicillin chrysogenum, Scopulariopsis ...

СПОСОБ ПОЛУЧЕНИЯ ДИПЕПТИДА (ВАРИАНТЫ)

... 1. Способ получения дипептида, включающий обеспечение воздействия белка, обладающего пролиниминопептидазной активностью, на эфир L-аминокислоты и L-аминокислоту с образованием дипептида, где белок, обладающий пролиниминопептидазной активностью, происходит из микроорганизма, принадлежащего к роду Pseudomonas. 2. Способ по п.1, где белок, обладающий пролиниминопептидазной активностью, происходит из Pseudomonas putida. 3. Способ по п.1 или 2, где эфир L-аминокислоты представляет собой один или несколько представителей, выбранных из группы, состоящей из эфира L-аланина, эфира глицина, эфира L-валина, эфира L-изолейцина, эфира L-метионина, эфира L-фенилаланина, эфира L-серина, эфира L-треонина, эфира L-глутамина, эфира L-тирозина, эфира L-аргинина, α-эфира L-аспарагиновой кислоты, β-эфира L-аспарагиновой кислоты, эфира L-лейцина, эфира L-аспарагина, эфира L-лизина, α,β-диметилового эфира L-аспарагиновой кислоты, и γ-эфира L-глутамина. 4. Способ по п.1 или 2, где L-аминокислота представляет собой ...

СПОСОБ ПОЛУЧЕНИЯ ПРЕДСТАВЛЯЮЩЕГО ИНТЕРЕС ПОЛИПЕПТИДА, ГИБРИДНАЯ ДНК (ВАРИАНТЫ), СЛИТЫЙ БЕЛОК (ВАРИАНТЫ)

Изобретение относится к биотехнологии и, в частности, к способам получения рекомбинантных полипептидов через промежуточную форму слитого белка. Предлагаемый способ предусматривает создание гибридной ДНК-конструкции, кодирующей белок, в котором две полипептидные последовательности связаны переходной областью, содержащей сайт распознавания Ig A-протеазой, трансформацию хозяйских клеток рекомбинантным вектором экспрессии, содержащим названную конструкцию, выделение полученного слитого белка, обработку его Ig A-протеазой при весовом соотношении фермента и субстрата от 1:1 до 100:1 и pH 6,5 - 8,5 и изолирование представляющего интерес полипептида. Получены, охарактеризованы и испытаны конкретные формы гибридных ДНК и слитых белков. Изобретение обеспечивает возможность получения представляющих интерес полипептидов в безметиониновой форме, а также позволяет повысить уровень их экспрессии и степень очистки. 11 с. и 8 з.п. ф-лы, 4 ил.

Системы экспрессии РНК рекомбинантного вируса ньюкаслской болезни и вакцины

... 1. Рекомбинантная молекула РНК, содержащая сайт связывания, специфичный для РНК-полимеразы вируса ньюкаслской болезни, и сигнальные последовательности, необходимые для NDV-опосредованной репликации и транскрипции, функционально присоединенные к гетерологичной последовательности РНК. 2. Рекомбинантная молекула РНК, содержащая сайт связывания для РНК-полимеразы вируса ньюкаслской болезни и сигнальные последовательности, необходимые для NDV-опосредованной репликации и транскрипции и функционально присоединенные к гену вируса ньюкаслской болезни, где указанная молекула РНК содержит мутацию, инсерцию или делецию. 3. Рекомбинантная молекула РНК по п.1 или 2, где указанный сайт связывания с полимеразой включает сайт связывания с полимеразой, локализованный в 3'- и 5'-некодирующей фланкирующей области РНК-генома вируса ньюкаслской болезни. 4. Рекомбинантная молекула РНК по п.3, где 3'- и 5'-некодирующая фланкирующая область имеет вирусную смысловую последовательность, показанную на фиг.1. 5. Рекомбинантная ...

Система экспрессии для фактора VIII

... 1. Способ получения клеток, экспрессирующих белок, обладающий прокоагулирующей активностью фактора VIII, включающий следующие последовательные этапы: a) получение клеток исключительно человеческого происхождения, b) взаимодействие клеток стадии а) с вектором в условиях, позволяющих вектору проникнуть в клетки, причем вектор включает селективный маркер и последовательность, кодирующую белок, обладающий прокоагулирующей активностью фактора VIII, оперативно связанную с промотором, c) отбор клеток стадии b) при помощи агента селекции и d) выделение из клеток, полученных на стадии с) отдельных клонов, экспрессирующих большие количества белка, обладающего прокоагулирующей активностью фактора VIII. 2. Способ по п.1, включающий далее стадию e) адаптации клонов стадии d) к росту в среде, свободной от белков плазмы. 3. Способ по п.1, где клетки стадии а) являются гибридами клеток лимфомы человека и клеток 293 S. 4. Способ по п.1, где клетки стадии а) являются гибридами клеток 2В8 (АТСС CRL-12569) ...

ВЫСОКОМОЛЕКУЛЯРНЫЕ ПОВЕРХНОСТНЫЕ БЕЛКИ НЕТИПИЧНЫХ ГЕМОФИЛЬНЫХ БАКТЕРИЙ

... 1. Выделенная и очищенная молекула нуклеиновой кислоты, кодирующая высокомолекулярный белок (НМW) НМW3 или НМW4 штамма нетипичных Haemophilus, характеризуемая: (а) последовательностью ДНК, показанной на фигуре 8 (SEQ ID : 7), кодирующей белок НМW3, обладающий аминокислотной последовательностью, приведенной на фиг.10 (SEQ ID :9), или (b) последовательностью ДНК, показанной на фигуре 9 (SEQ ID :8), кодирующей белок НМW4, обладающий аминокислотной последовательностью, приведенной на фиг.10 (SEQ ID :10). 2. Выделенная и очищенная молекула нуклеиновой кислоты, кодирующая высомоколекулярный белок (НMW) штамма нетипичных Haemophilus, которая выбрана из группы, состоящей из: (а) последовательностью ДНК, как показано на любой одной из фиг.8 и 9 (SEQ ID :7 и 8); (b) последовательностью ДНК, кодирующей аминокислотную последовательность, которая показана на фиг.10 (SEQ ID :9 и 10); или (с) последовательностью ДНК, кодирующей высокомолекулярный белок штамма нетипичных Haemophilus, которая гибридизует ...

СВЯЗЫВАЮЩИЕ TNF-α БЕЛКИ

... 1. Антитело или его антиген-связывающая часть, содержащие SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39; SEQ ID NO: 40; SEQ ID NO: 41; SEQ ID NO: 42; SEQ ID NO: 43; SEQ ID NO: 44; SEQ ID NO: 45 или SEQ ID NO: 46.2. Антитело или его антиген-связывающая часть, способные связывать TNF-α, включающие по меньшей мере одну CDR, содержащую остатки 31-35 SEQ ID NO: 31 (CDR-H1), остатки 50-66 SEQ ID NO: 31 (CDR-H2), остатки 99-113 SEQ ID NO: 31 (CDR-H3), остатки 24-34 SEQ ID NO: 32 (CDR-L1), остатки 50-56 SEQ ID NO: 32 (CDR-L2) или остатки 89-97 SEQ ID NO: 32 (CDR-L3).3. Связывающий белок по п.2, где связывающий белок содержит по меньшей мере три CDR.4. Связывающий белок по п.3, где по меньшей мере три CDR содержат набор вариабельных доменов CDR из:(a) остатков 31-35 SEQ ID NO: 31 (CDR-H1), остатков 50-66 SEQ ID NO: 31 (CDR-H2) и остатков 99-113 SEQ ID NO: 31 (CDR-H3); или(b) остатков 24-34 SEQ ID NO: 32 (CDR-L1 ...

ВАКЦИНА ВИРУСА ГРИППА

... 1. Конъюгат пептид М2-белок, включающий множество пептидов, имеющих аминокислотную последовательность, полученную из внеклеточного домена белка М2 вируса гриппа типа А, где указанное множество пептидов ковалентно связано с поверхностью белка-носителя, и каждая указанная связь образована одним концом пептида и реакционноспособным сайтом на поверхности указанного белка, где белок-носитель выбирают из группы, состоящей из белкового комплекса наружной мембраны Neiserria meningitidis, анатоксина столбняка, поверхностного антигена гепатита В, гемоцианина лимфы улитки, капсидного белка ротавируса и белка L1 VLP вируса папилломы коровы или человека, или его фармацевтически приемлемая соль. 2. Конъюгат по п.1, где аминокислотную последовательность пептида выбирают из группы состоящей из последовательностей, представленных SEQ ID NO: 1, 2, 10 и 39. 3. Конъюгат по п.2, где указанный пептид имеет последовательность, представленную SEQ ID NO: 39. 4. Конъюгат по п.1, где указанный белок-носитель представляет ...

СПОСОБ ПОЛУЧЕНИЯ ПОЛИПЕПТИДОВ

... 1. Способ получения полипептида, гетерологичного для E. coli, предусматривающий (a) культивирование клеток E. coli, содержащих кодирующую полипептид нуклеиновую кислоту, в культуральной среде при подпитке культуральной среды транспортабельным органофосфатом, так что нуклеиновая кислота экспрессируется, и (b) выделение полипептида из клеток. 2. Способ по п.1, где органофосфат представляет собой глицерофосфат. 3. Способ по п.2, где глицерофосфат представляет собой альфа-глицерофосфат или бета-глицерофосфат или их смесь. 4. Способ по п.3, где глицерофосфат представляет собой смесь глицерол-2-фосфата и глицерол-3-фосфата или представляет собой глицерол-3-фосфат. 5. Способ по любому из пп.1-4, где культивирование проводят во встряхиваемой колбе или ферментере. 6. Способ по любому из пп.1-4, где полипептид выделяют из цитоплазмы, периплазмы или культуральной среды клеток. 7. Способ по любому из пп.1-4, где экспрессию нуклеиновой кислоты регулируют индуцибельным промотором. 8. Способ по п.7, где ...

Способ приготовления питательной среды

Способ получения инсулина человека

Способ получения человеческого инсулина

Enzymatische Hydrolyse von Proteinen

Enzymatische Hydrolyse von Proteinen

VERFAHREN ZUR HERSTELLUNG REKOMBINANTER, BIOLOGISCH AKTIVER POLYPEPTIDE

VERFAHREN ZUR SELEKTIVEN SPALTUNG VON FUSIONSPROTEINEN.

Prodn. of polypeptides of formula (Gly)q-(X1---X3) (I) is effected by clearing a fusion protein of formula (Y1---Ym)-(Gly) p+q-(X1---Xn) (II) with a polyglycine-specific endoprotease (III): (In the formulae X and Y are natural amino acids; m and n are integers greater than 1; p and q are integers greater than O such that p+q = 2-100, or p may be O and p+q may be less than 2 when Ym= Gly, or q may be O and p+q may be less than 2 when X1=Gly.) ...

AUS CHAPERONIN-POLYPEPTIDEN GEBILDETE GEORDNETE BIOLOGISCHE NANOSTRUKTUREN

VERFAHREN ZUR EXTRAKTION, REINIGUNG UND ENZYMATISCHEN VERÄNDERUNG VON SOJA 7S GLOBULIN ALPHA' UNTEREINHEIT ZUR VERWENDUNG ALS HYPOCHOLESTEROL-MISCHER-WIRKSTOFF

HERSTELLUNGSVERFAHREN FÜR ITURIN A UND DESSEN HOMOLOGE

"DEATH-DOMAIN"-ENTHALTENDER REZEPTOR 4 (DR4), EIN MITGLIED DER TNF-REZEPTOR SUPERFAMILIE, WELCHER AN TRAIL (APO-2L) BINDET

UL16-BINDUNGSPROTEIN 4

Methode zur Herstellung von menschlichen Wachstumshormonen

PHENOL OXYDIERENDE ENZYME UND DEREN VERWENDUNG

Reinigungsverfahren auf Grundlage der Bindung von Enzymen and markierte Peptide

Verfahren zur Herstellung eines bluddrucksenkenden Mittel enthaltenden ein Dipeptid als wirksames Bestandteil

Stable storage of proteins

A method of stably storing a protein, the method comprising applying a protein to be stored to a substrate which has been treated with a polyhydric compound and dried, wherein the amount of the polyhydric compound present in the substrate is sufficient to stabilise the protein, and wherein the substrate does not consist of glass. The substrate preferably comprises cellulose fibres and the polyhydric compound is selected from the group consisting of: a PVA, glycerol, sucrose, carrageenan, xanthum gum and pectin.

LOW PHENYLALANINE PLASTEINS

... 1498923 Plasteins FUJI OIL CO Ltd 15 Jan 1976 [16 Jan 1975] 01601/76 Heading C3H A low-phenylalanine content plastein is made by (a) hydrolysing an albumin on a globulin protein with an endopeptidase specific for the aromatic amino acid constituents of the protein and subsequently with an exopeptidase to give a hydrolysate, (b) subjecting the resulting hydrolysate to a gel filtration to divide it into two fractions, the one containing the aromatic amino acid components and the other containing substantially no free or combined phenyl alanine, i.e. up to 1À9% by weight and (c) subjecting the latter fraction to a plastein synthesis. The protein source may be meat, fish, egg, oil-seed or single cell. Examples describe the hydrolysis of fish protein using pepsin and a mixture of carboxypeptidase A and leucineaminopeptidase. Elution of the hydrolysate through a dextran bed yielded a product low in phenylalanine content. A complete amino acid breakdown and plastein synthesis are reported.

Method for production of therapeutically pure granulocyte macrophage-colony stimulating factor

Prion protein binding materials and methods of use

Method for production of therapeutically pure granulocyte macrophage-colony stimulating factor

Prion protein binding materials and methods of use

Methods and compositions for immunization against HIV

Method for production of therapeutically pure granulocyte macrophage-colony stimulating factor

An over-expressing homologous antigen vaccine and a method of making the same.

Process of solubilization and hydrolysis of isolated protein.

Methods and compositions for immunization against HIV

Methods and compositions for immunization against HIV

Prion protein binding materials and methods of use

ON TGF BETA 1 RESPONDING MARROW CELLS

HUMAN ALPHA ENDOKIN

REMODELLIERUNG OF GLYCOPROTEINEN USING ENDOGLYCANASEN

COMPOSITIONS FOR THE SYSTEMIC ADMINISTRATION OF SEQUENCES, FOR KNOCHENMORPHOGENESE PROTEINS CODING

T1R-GESCHMACKSREZEPTOREN AND YOUR CODING GENES

PROCEDURE FOR REMOVING FROM THE N-TERMINAL ALANINE REMAINDERS WITH AEROMONAS AMINOPEPTIDASE

PROCEDURES FOR THE PRODUCTION ONE COMPLETELY ENZY MATISCH HYDROLYZED PRODUCT OF GRAIN IN PARTICULAR FROM FULL GRAIN

ALKALINE PROTEASE VARIANTS AND WASH AND CLEANING AGENTS CONTAINING THESE ALKALINE PROTEASE VARIANTS

VERWENDUNG VON CHYMOTRYPSIN ZUM UNWIRKSAMMACHEN DES PRAEKALLIKREIN-AKTIVATORS

PROCEDURE FOR DEACYLIERUNG OF LIPOPEPTIDEN

CHIMÄRE HUMAN ONES PAPILLOMAVIRUS (HPV) L1 of MOLECULES AND THEIR USES

POLYP CEPT ID DISPLAY LIBRARIES AND PROCEDURES FOR THE PRODUCTION AND USE OF IT

MOLECULAR IDENTIFICATION OF ASPERGILLUSSPEZIES

REKONSTITUIERTER HISTON METHYLTRANSFERASE COMPLEX AND PROCEDURE FOR THE IDENTIFICATION OF MODULATORS FOR IT

IN TASTE RECEPTOR CELLS OF EXPRIMERTER TRANSIENT RECEPTOR POTENTIAL CHANNEL TRP8

NUCLEOTIDE AND PROTEIN SEQUENCES OF A SERRATE OF GENE FROM VERTEBRATE ANIMALS AND WHEREUPON BASING PROCEDURE

FGF-5-BINDENDE AND GETRÄGERTE PEPTIDE

POLYP CEPT ID SPLITTING PROCESS

USE FROM TRYPTOPHANREICHEN PEPTIDEN WHEY PROTEIN HYDROLYSATES FOR THE TREATMENT OF PREDOMINANCE AND OBESITY

NATURAL IGM ANTIKÖRPER ONES AND INHIBITORS OF IT

THE POTASSIUM CHANNEL KCNQ5

PROCEDURE FOR THE PRODUCTION OF HYALURONAN IN A REKOMBINANTEN LANDLORD CELL

USE OF A NUCLEOTIDE SEQUENCE, WHICH CODES FUR ENZYME I-SCIE

ADJUSTED TRANSKRIPTION FROM PURPOSEFUL GENES AND OTHER BIOLOGICAL EVENTS

PHOSPHATE-LIMITED INDUCE-CASH PROMOTER AND BACILLUS EXPRESSIONSSYSTEM ONE

GENES OF CORYNEBACTERIUM GLUTAMICUM CODING FOR METABOLIC PROTEINS

PROCEDURE AND DNA KONSTRUKTE FOR THE PRODUCTION OF POLYPEPTIDEN WITH HIGH YIELD

MANIPULATED STENCILS AND YOUR USE WITH SINGLE PRIMER AMPLIFICATION

UL16-BINDUNGSPROTEIN 4

GLYCOPROTEINSYNTHESE

REKOMBINANTES, 250 KDA LARGE ANTIGEN FROM SPOROZOITEN/MEROZOITEN NUCLEIC ACIDS CODING OF EIMERIA MAXIMA AND YOUR USES

PHARMACEUTICAL CONNECTION AND METHODS TO TREATMENTS OF HUMAN CARCINOMAS OF MEANS ARGININ WITHDRAWAL

PROCEDURE FOR THE EVALUATION OF RIBONUKLEOTIDSEQUENZEN

FETTALKOHOLOXIDASE GENES AND - PROTEINS FROM CANDIDA TROPICALIS AND THUS CONNECTION PROCEDURE

ELONGASEGENE AND THEIR USES

REKOMBINANTE ADENOVIREN WITH APE ADENOVIRUS PROTEINS AND USE OF IT.

LACCASEN FOR BIOLOGICAL BLEACHING OF CELLULOSE

PROCEDURE FOR EXPRESSION AND CLEANING OF IMMUNOTOXINEN

PROCEDURE FOR THE UNCOUPLING OF THE REACTION YARDSTICK AND THE PROTEIN SYNTHESIS YIELD IN THE INTERMITTENT MODE

EXPANSION OF THE EUKARYOTI GENETIC CODE

MUTATED BACILLUS LICHENIFORMIS LANDLORD CELL

PROCEDURE FOR THE PRODUCTION OF KOLLAGEN HYDROLYSATES

PREVENTS FROM SPLITTING OF N-TERMINAL AMINO ACIDS BY ENDOGENOUS AMINOPEPTIDASEN DURING THE EXPRESSION OF STRANGE GENES IN BACTERIA

PROCEDURE FOR THE PRODUCTION OF A PEPTIDS OF MEANS OF A HILFSPEPTIDS

PROCEDURE FOR THE PRODUCTION OF CS6 ANTIGEN VACCINE FOR THE TREATMENT, PREVENTION OR INHIBITION OF ENTEROTOXIGENER ESCHERICHIA COLI INFECTIONS

PROCEDURE FOR THE PRODUCTION OF KOLLAGEN HYDROLYSATES

MASS-SPECTROMETRIC ANALYSIS OF BIO POLYMERS

HIGH BONE MASS the GENE ON 11Q13.3

BACTERIA TRUNKS OF THE GENUS BACILLUS, PHENOTYPISCH LACTOBACILLUS AEHNLICH, PROCEDURE FOR CULTIVATION AND USE

Synthesis of Site Specifically-Linked Ubiquitin

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 ...

MODIFIED SAK GENE FOR THE PRODUCTION OF RECOMBINANT PROTEINS

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) ...

METHOD FOR PRODUCING PEPTIDE

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 ...

Extracellular Matrix-Derived Gels and Related Methods

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 ...

C-TERMINAL MODIFICATION OF POLYPEPTIDES

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 ...

Glycosylation of Molecules

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 ...

WHOLE EGG PROTEIN PEPTIDES, PREPARATION METHOD AND USE THEREOF

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 ...

HYDROPHOBIZED PROTEIN HYDROLYSATE

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 , ...

METHOD FOR PREPARING ACTIVE PEPTIDES FROM CORN GERM PROTEINS

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 ...

PLANT PROTEIN HYDROLYSATES

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 ...

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

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 ...

COMPOSITIONS AND METHODS OF PRODUCING ENTEROKINASE IN YEAST

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 ...

METHOD FOR THE PREPARATION OF SURFACTANT PEPTIDES

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 ...

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

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 ...

ACID-CLEAVABLE LINKERS EXHIBITING ALTERED RATES OF ACID HYDROLYSIS

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 ...

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

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 ...

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

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 ...

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

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 , ...

EFFICIENT PRODUCTION OF PEPTIDES

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 , ...

METHODS FOR PRODUCTION AND PURIFICATION OF POLYPEPTIDES

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 ...

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

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 ...

EXPRESSION AND LARGE-SCALE PRODUCTION OF PEPTIDES

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 ...

Sortase-mediated protein purification and ligation

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 ...

Method for Producing a Protein Hydrolysate

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 ...

SYNTHETIC PEPTIDES AND ENZYMATIC FORMATION OF INTRACELLULAR HYDROGELS

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 ...

ON-COLUMN ENZYMATIC CLEAVAGE

Herein is reported a method for obtaining a polypeptide by an immobilized metal ion affinity chromatography from a pro-polypeptide that comprise at its N- or C-terminus an metal ion affinity chromatography tag and a protease cleavage site comprising the step of recovering the polypeptide from the immobilized metal ion affinity chromatography column by incubating the bound pro-polypeptide with a protease, whereby the immobilized metal ion affinity chromatography material has been washed at least once with an urea solution. 1. A method for producing a polypeptide from a pro-polypeptide , whereby the pro-polypeptide comprises at its N- or C-terminus a metal ion affinity chromatography tag and a protease cleavage site located between the tag and the polypeptide , by an on-column enzymatic cleavage of the protease cleavage site on an immobilized metal ion affinity chromatography column comprising the following steps:denaturing the pro-polypeptide bound to the metal ion affinity chromatography material,renaturing the pro-polypeptide bound to the metal ion affinity chromatography material, andincubating the bound pro-polypeptide with a protease and thereby producing the polypeptide.2. A method for producing a polypeptide from a pro-polypeptide , whereby the pro-polypeptide comprises at its N- or C-terminus a metal ion affinity chromatography tag and a protease cleavage site located between the tag and the polypeptide , by an on-column enzymatic cleavage of the protease cleavage site on an immobilized metal ion affinity chromatography column comprising the following steps:contacting the bound pro-polypeptide with a solution comprising a denaturing agent,optionally contacting the bound pro-polypeptide with a solution comprising urea or a urea derivatives if the solution comprising a denaturing agent employed in the previous step was free of urea or a urea derivative or contacting the bound pro-polypeptide with a solution comprising urea or a urea derivatives if the solution ...

METHOD FOR THE PREPARATION OF LOW MOLECULAR WEIGHT PORCINE LYMPHO-RETICULAR POLYPEPTIDES

A method for preparation of low molecular weight porcine lympho-reticular polypeptides. The method comprises the enzymatic hydrolysis of a source of protein, wherein the source of protein comprises a blend of porcine liver and porcine spleen, with an enzyme having proteolytic activity and an enzyme having amylase activity. 1. A method for preparation of low molecular weight porcine lympho-reticular polypeptides , comprising:enzymatic hydrolysis of a source of protein, wherein the source of protein comprises a blend of porcine liver and porcine spleen, with an enzyme having proteolytic activity and an enzyme having amylase activity.2. The method of claim 1 , wherein the blend of porcine liver and spleen claim 1 , is present at a ratio of about 2:8 to 8:2 respectively.3. The method of claim 1 , wherein the enzyme having proteolytic activity is selected from the group consisting of pancreatin claim 1 , papain claim 1 , pepsin claim 1 , or mixtures thereof.4. The method of claim 1 , wherein the enzyme having amylase activity is diastase.5. The method of claim 3 , wherein the papain is used at an amount ranging from 0.5 to 2.0 ounces per eleven pounds of the blend of porcine liver and spleen.6. The method of claim 3 , wherein the pepsin is used at an amount ranging from 1.5 to 3.5 ounces per eleven pounds of porcine glands.7. The method of claim 3 , wherein the pepsin is provided in the form of anhydrous pepsin.8. The method of claim 3 , wherein the pancreatin is used at an amount ranging from 0.05 to 1.0 ounces per eleven pounds of fresh porcine glands.9. The method of claim 4 , wherein the diastase is used at an amount ranging from 0.1 to 1.5 ounces per eleven pounds of fresh porcine glands.10. The method of claim 1 , further comprising a cleansing step in an aqueous surfactant solution containing benzalkonium chloride.11. The method of claim 1 , wherein the enzymatic hydrolysis has a pH range of between 4.5 to 5.5.12. The method of claim 1 , wherein the pH range is ...

MODIFIED ENTEROKINASE LIGHT CHAIN

The present invention is related to novel mammalian enterokinase analogues such as mammalian enterokinase light chain analogues and methods of making such. Also described herein is a method for cleaving proteins having an enterokinase cleavage site. 1. A bovine enterokinase light chain analogue comprising at least one substitution in position 134 and/or 135 from hydrophobic to a hydrophilic charged amino acid(s).2. The bovine enterokinase light chain analogue according to claim 1 , further comprising a substitution in position 112.3. The bovine enterokinase light chain analogue according to claim 1 , wherein the hydrophilic charged amino acid(s) are one or more amino acids selected from the group consisting of: lysine claim 1 , arginine claim 1 , glutamic acid and aspartic acid.4. The bovine enterokinase light chain analogue according to claim 1 , wherein the enterokinase light chain to be mutated is SEQ ID NO:1.5. A method for obtaining improved solubility in a renaturation process of an enterokinase light chain analogue comprising the step of mutating one or more hydrophobic amino acids of wild type bovine enterokinase light chain to hydrophilic amino acids and optionally mutating other amino acids of wild type bovine enterokinase light chain claim 1 , wherein the hydrophobic amino acids subject to mutation are present on the surface of folded wild type bovine enterokinase light chain.6. The method according to claim 5 , wherein the hydrophobic amino acid(s) to be mutated are selected from the group consisting of: I claim 5 , V claim 5 , L claim 5 , M claim 5 , W claim 5 , F claim 5 , and A.7. The method according to claim 5 , wherein the hydrophilic amino acid(s) are selected from the group consisting of: Lysine claim 5 , arginine claim 5 , glutamic acid and aspartic acid.8. The method according to claim 5 , wherein the hydrophobic amino acid(s) to be mutated are in one or more positions selected from the group consisting of: position 11-14 (amino acids AWPW) ...

DIPEPTIDYL PEPTIDASE-4 INHIBITOR

The present invention aims to provide a DPP-4 inhibitor that is obtained by using a food as a raw material and that is suitable for oral ingestion from the viewpoints of flavor and absorbability, and a composition for the prevention and/or amelioration of diabetes which contains the DPP-4 inhibitor. The present invention provides a DPP-4 inhibitor obtained by treating an azuki bean or a kidney bean with a microorganism or a proteolytic enzyme produced by the microorganism. In particular, a preferable DPP-4 inhibitor can be obtained by hydrolyzing an azuki bean with a koji mold or a proteolytic enzyme derived from the koji mold to fragment a protein in the azuki bean. 1. A method for the prevention and/or amelioration of diabetes , comprising administrating to a subject the protein hydrolysate derived from an azuki bean or a kidney bean , which is a DPP-4 inhibitor and which is obtained by the method comprising treating an azuki bean , a kidney bean or a treated bean thereof , suspended or dissolved in water with a proteolytic enzyme produced by a koji mold.2Aspergillus oryzaeAspergillus sojae.. The method for the prevention and/or amelioration of diabetes according to claim 1 , wherein the koji mold is or3. The method for the prevention and/or amelioration of diabetes according to claim 1 , wherein the proteolytic enzyme comprises a peptidase.4. The method for the prevention and/or amelioration of diabetes according to claim 1 , wherein the proteolytic enzyme has X-prolyl dipeptidyl aminopeptidase activity.5. A method of producing a protein hydrolysate derived from an azuki bean or a kidney bean claim 1 , which is a DDP-4 inhibitor claim 1 , comprising treating an azuki bean claim 1 , a kidney bean or a treated bean thereof claim 1 , suspended or dissolved in water with a proteolytic enzyme produced by a microorganism.6. The method of producing protein hydrolysate derived from an azuki bean or a kidney bean according to claim 5 , which is a DPP-4 inhibitor claim 5 , ...

PREPRATION METHOD AND APPLICATION OF ANTI-INFLAMMATORY KIDNEY PROTECTING CLAM PEPTIDE

A preparation method and application of an anti-inflammatory and kidney-protecting clam peptide are disclosed. The method includes the steps of: cleaning and crushing the whole meat of a HonDau clam to obtain serous fluid, adding compound protease accounting for 0.1-0.3% of the weight of the serous fluid, carrying out enzymolysis, centrifugation, membrane separation and purification, and spray drying to obtain clam peptide powder. The clam peptide is prepared by adopting the HonDau clam, and is used for relieving the inflammatory reaction of an organism and the damage of renal function caused by hypertension, and is further applied to the inflammation and the damage of kidney caused by other diseases and health care foods with related functions. 1. A method for preparing an anti-inflammatory and kidney-protecting clam peptide , comprising:cleaning and mashing a whole clam meat to obtain slurry;adding compound protease accounting of 0.1-0.3% of a weight of the slurry; andperforming enzymolysis, centrifugation, membrane separation and purification, and spray drying to obtain clam peptide powder.2. The method of claim 1 , wherein the compound protease comprises neutral protease claim 1 , alkaline protease and flavor protease; and the weight ratio of the neutral protease to the alkaline protease to the flavor protease is 2:1:1.3. The method of claim 1 , wherein the weight ratio of the clam meat to water is 1:1 to 1:3 during the enzymolysis.4. The method of claim 1 , wherein during the enzymolysis claim 1 , the clam peptide is subjected to enzymolysis at a temperature of 50-60° C. for 4-6 hours.5. The method of claim 1 , wherein during the centrifugation claim 1 , a centrifugal speed of the clam peptide in the production process is 16000 R/min.6. The method of claim 1 , wherein during the membrane separation and purification claim 1 , the clam peptide enzymolysis solution is subjected to microfiltration-ultrafiltration-nanofiltration membrane filtration claim 1 , and the ...

COMPOSITIONS AND METHODS FOR PRODUCING CLOSTRIDIAL COLLAGENASES

The present invention provides a method for producing a drug product comprising a combination of highly purified collagenase I and collagenase II from . The method utilizes an improved medium for the cultivation of which includes a non-meat-derived (i.e., non-mammalian) peptone or vegetable peptone. The method includes one or more of: (1) reducing glucose content in the meat-free or vegetable-derived media; and (2) increasing the salt concentration in the meat-free or vegetable-derived media. Also provided is a drug product which includes collagenase I and collagenase II at an optimized fixed mass ratio, and which has a purity of greater than at least 95%. 1Clostridium histolyticum. A medium for the fermentation of comprising:a. a peptone selected from the group consisting of Oxoid VG100 Vegetable Peptone made from pea, Oxoid VG200 Vegetable Peptone phosphate broth, BBL Phytone Peptone, and BD Difco Select Phytone;b. yeast extract; andc. less than about 5 g/L glucose;wherein the pH of said medium is 7.5-7.9.2. The medium of claim 1 , containing greater than about 5 g/L salt.3. The medium of claim 2 , containing greater than about 7.5 g/L salt.4. The medium of claim 2 , wherein the salt is one or more of potassium phosphate claim 2 , dipotassium phosphate claim 2 , sodium phosphate claim 2 , disodium phosphate claim 2 , sodium chloride claim 2 , potassium chloride claim 2 , calcium chloride claim 2 , or magnesium sulfate.5. The medium of claim 1 , further comprising one or more vitamins selected from the group consisting of ferrous sulfate claim 1 , riboflavin claim 1 , niacin claim 1 , calcium pantothenate claim 1 , pimelic acid claim 1 , pyridoxine hydrochloride and thiamine hydrochloride.6Clostridium histolyticum. A medium for the fermentation of comprising:a. a peptone selected from the group consisting of Oxoid VG100 Vegetable Peptone made from pea, Oxoid VG200 Vegetable Peptone phosphate broth, BBL Phytone Peptone, and BD Difco Select Phytone;b. yeast extract; ...

proNGF mutants and uses thereof for the preparation of NGF mutants

Pro-NGF—Nerve Growth Factor—mutants and their uses for the production of NGF mutants are disclosed. Also disclosed are useful new process to optimize and improve the purification of NGF P61SR100E, a therapeutic protein. The invention further provides a mutant pro-NGF wherein the protease cleavage site RASKRB is substituted at least at position RA and K by a non-basic amino acid, and the mutant pro-NGF further includes at least one mutation selected from P165S or R204E. 1. A pro-NGF mutant wherein the protease cleavage site RSKRis substituted at least at position RA and K by a non-basic amino acid said pro-NGF mutant further comprising at least one mutation selected from P165S or R204E.2. The pro-NGF mutant according to wherein the non-basic amino acid is selected from the group consisting of Alanine claim 1 , Glycine claim 1 , Valine claim 1 , Serine claim 1 , Threonine claim 1 , Methionine claim 1 , Tyrosine claim 1 , Histidine claim 1 , Asparagine claim 1 , Aspartic Acid claim 1 , Glutamine claim 1 , Glutamic Acid claim 1 , Phenylalanine claim 1 , Isoleucine claim 1 , Leucine claim 1 , Tryptophan claim 1 , Cysteine claim 1 , and Proline.3. The pro-NGF mutant according to wherein the protease cleavage site has the sequence VSAR.4. The pro-NGF mutant according to comprising both mutations P165S and R204E.5. The pro-NGF mutant according to for use in a method to produce a NGF mutant.6. The pro-NGF mutant according to for use in a method to produce a NGF mutant claim 5 , said NGF mutant comprising at least one of the mutation selected from NGF P61S or R100E.7. A method for the production of a NGF mutant comprising the steps of incubating the pro-NGF mutant as defined in with trypsin.8. The method according to wherein the incubation time is about 24 hours.9. The method according to wherein the incubation temperature is between about 4° to about 16° C.10. The method according to wherein the weight ratio trypsin:pro-NGF is 1:5000 claim 7 , the incubation temperature is ...

Method of Preparation of SOD Hydrolysates Powder

A method for preparing powdery superoxide dismutase (SOD) hydrolysates. SOD is hydrolyzed by cellulase and then further hydrolyzed with a solution containing a mixture of proteases. Organic citric acid is added in the SOD hydrolysates solution, then the solution is freeze-dried to obtain the powdery SOD hydrolysates. 1. A method for preparing powdery SOD hydrolysates , comprising:(a) obtaining a solution containing polypeptides by hydrolyzing an SOD by a cellulase;(b) obtaining a composite enzyme solution;(c) mixing the solution containing polypeptides obtained in (a) with the composite enzyme solution obtained in (b) to thereby obtain a solution of SOD hydrolysates;(d) adding citric acid to the solution of SOD hydrolysates obtained in (c) to thereby form a further mixture; and(e) freeze-drying the further mixture obtained in (d) to obtain powdery SOD hydrolysates.2. The method of claim 1 , wherein obtaining the composite enzyme solution comprises:A. Obtaining a solution containing a protein;B. adjusting the pH of the solution to about 9.0;C. adding a basic protease to the solution, heating the solution, cooling the solution, and repeating the heating and cooling until the pH of the solution stabilizes at about 7;D. adjusting the pH of the solution obtained from step C to about 8.0;E. adding a neutral protease to the solution, heating the solution, cooling the solution, and repeating the heating and cooling until the pH of the solution stabilizes at about 7;F. adjusting the of the solution obtained from step E to about 3.5;G. adding an acidic protease to the solution, heating the solution, cooling the solution, and repeating the heating and cooling until the pH of the solution stabilizes at about 2-3.3. The method of claim 1 , wherein the SOD is derived from a plant.4. The method of claim 1 , wherein the SOD is an SOD derived from one of corn claim 1 , garlic claim 1 , onion claim 1 , wheat claim 1 , or combinations thereof.5. The method of claim 2 , wherein the ...

METHOD FOR PRODUCING A RECOMBINANT PROTEIN OF INTEREST

Disclosed is a method for producing a recombinant protein of interest which is characterised by the following steps: 1. Method for producing a recombinant protein of interest characterised by the following steps:{'sup': pro', 'pro', 'pro, '(a) providing a first part of an Nautoprotease and providing a second part of an Nautoprotease, wherein said second part is fused by a peptidic bond to a protein of interest but said second part alone does not exhibit a proteolytic activity, and wherein complementation of said first part with the second part forms an autoproteolytically active Nautoprotease,'}{'sup': pro', 'pro', 'pro', 'pro', 'pro, '(b) contacting the first part of the Nautoprotease with the second part of the Nautoprotease so that an autoproteolytically active Nautoprotease is formed and the protein of interest fused by the peptidic bond to the second part of the Nautoprotease is proteolytically cleaved off the second part of the Nautoprotease at the peptidic bond, and'}(c) recovering the protein of interest.2E. coli. Method according to claim 1 , characterized in that the first and/or second part of the Nautoprotease were generated in a recombinant production system claim 1 , preferably in a prokaryotic host cell claim 1 , especially in host cells.3. Method according to claim 1 , characterized in that the first part of the Nautoprotease comprises amino acid 22 to 30 of Nautoprotease claim 1 , preferably amino acid 22 to 75 claim 1 , more preferred amino acid 22 to 112 claim 1 , 22 to 106 claim 1 , 1 to 106 claim 1 , 1 to 75 or 1 to 30 claim 1 , especially amino acid 1 to 112.4. Method according to claim 1 , characterized in that the second part of the Nautoprotease comprises amino acid 31 to 168 of Nautoprotease claim 1 , preferably amino acid 76 to 168 claim 1 , especially amino acid 113 to 168 and 107 to 168.5. Method according to claim 1 , characterized in that either said first part or said second part of the Nautoprotease is provided in immobilised form on ...

METHOD FOR PRODUCING A RECOMBINANT PROTEIN OF INTEREST

Disclosed is a method for producing a recombinant protein of interest, the method being characterised in by the following steps: 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 inclusion bodies,{'sup': 'pro', '(c) allowing the fusion protein to be cleaved by the Nautoprotease moiety under chaotropic conditions, wherein the recombinant protein of interest is cleaved from the fusion protein and wherein the recombinant protein of interest is not yet renatured or simultaneously renatured, and'}(d) recovering the protein of interest, optionally including a renaturing step for the protein of interest.2E. coli. Method according to claim 1 , characterized in that the inclusion bodies were generated in a recombinant production system claim 1 , preferably in a prokaryotic host cell claim 1 , especially in host cells.3. Method according to or claim 1 , characterized in that the conditions in step (b) correspond to a urea concentration of more than 5 M claim 1 , preferably more than 6 M claim 1 , especially more than 7.5 M.4. Method according to claim 1 , characterized in that the chaotropic conditions in step (c) correspond to a urea concentration of 1.4 to 5 M claim 1 , preferably from 2 to 5 M claim 1 , especially from 2 to 4 M.5. Method according to claim 1 , characterized in that the Nautoprotease moiety has a cleavage rate at 2.5 M urea of at least 20% claim 1 , preferably at least 30% claim 1 , especially at least 40%.6. Method according to claim 1 , characterized in that the Nautoprotease moiety has a sequence selected from the group consisting of SEQ ID nos. 1 or 2.7. Method according to claim 1 , characterized in that the Nautoprotease moiety has a sequence wherein the interferon regulating factor 3 binding site is deleted.8. Method according to claim 1 , ...

Leucine-Rich Peptide Compositions and Methods for Isolation

Disclosed are compositions comprising isolated peptides having a leucine content of from about 12 to about 40 weight percent. Also disclosed is a method for isolating leucine-rich peptides from protein sources such as bovine whey and methods of use for these peptides to provide beneficial effects in a human and/or animal such as increasing blood flow, decreasing blood pressure, increasing muscle mass, improving cognitive function, improving cardiovascular function, etc. 1. A composition comprising isolated peptides having a leucine content of from about 12% to about 40% by weight and a molecular weight of from about 200 to about 4 ,000.2. The composition of wherein the leucine content of the isolated peptides is from about 12% to about 30% by weight.3. The composition of wherein the leucine content of the isolated peptides is from about 15 to about 20 weight percent.4. The composition of wherein the leucine content of the isolated peptides is from about 15 to about 30 weight percent.5. The composition of wherein the leucine content of the isolated peptides is from about 20 to about 30 weight percent.6. The composition of wherein the molecular weight of the isolated peptides is from about 200 to 1 claim 1 ,000.7. The composition of wherein the molecular weight of the isolated peptides is from about 400 to 1 claim 1 ,000.8. The composition of wherein the protein source is chosen from the group consisting of legumes claim 1 , fish claim 1 , meat claim 1 , milk claim 1 , egg claim 1 , corn claim 1 , wheat gluten claim 1 , maize claim 1 , and combinations thereof.9. The composition of wherein the protein source is chosen from the group consisting of bovine milk claim 1 , bovine whey claim 1 , whey protein concentrates claim 1 , whey protein isolates claim 1 , and combinations thereof.10. A peptide composition comprising peptides having a leucine content of from about 12 to about 40 weight percent and a molecular weight from about 200 to about 4 claim 1 ,000 claim 1 , the ...

ACTIVE PEPTIDE FOR INHIBITING AMPA RECEPTOR AND PREPARATION METHOD AND USE THEREOF

Provided is an active peptide for inhibiting an AMPA receptor and a preparation method and use thereof. The method for preparing the active peptide comprises the following steps: 1) soaking a salmon skin and crushing, adding water and beating, and then adjusting pH to 6.5-7.5; 2) subjecting to a first enzymolysis using a neutral protease; 3) subjecting to a second enzymolysis using papain enzyme and then inactivating enzyme; and 4) centrifuging the enzymatic hydrolysate, and then subjecting the centrifuged supernatant to membrane filtration, concentration and decoloration, to prepare the active peptide. The active peptide contains a tetrapeptide with an amino-acid sequence of Glu-Gly-Ala-Arg. The tetrapeptide has good solubility, can selectively inhibit neuronal synaptic transmission caused by an AMPA receptor, and has a significant antiepileptic effect. 1. A method for preparing an active peptide , comprising the following steps:1) soaking a salmon skin and crushing, adding water and beating, and then adjusting pH to 6.5-7.5, to obtain a slurry;2) subjecting the slurry to a first enzymolysis by using a neutral protease, to obtain a first enzymatic hydrolysate;3) subjecting the first enzymatic hydrolysate to a second enzymolysis by using papain enzyme and then inactivating enzyme, to botain a second enzymatic hydrolysate; and4) centrifuging the second enzymatic hydrolysate, and then subjecting the centrifuged supernatant to membrane filtration, concentration and decoloration, to prepare the active peptide;wherin the active peptide contains a tetrapeptide with an amino-acid sequence of Glu-Gly-Ala-Arg.2. The method according to claim 1 , wherein an alkaline solution with a mass content of 0.1-0.5% is used to soak the salmon skin claim 1 , a mass/volume ratio of the salmon skin to the alkaline solution is controlled to 1:(2-4) claim 1 , and soaking time is 5-20 h.3. The method according to claim 1 , wherein the amount of the neutral protease is 50-500 U/g claim 1 , ...

METHOD FOR PREPARATION OF A RECOMBINANT PROTEIN FROM A PRECUSOR

The method for preparation of a recombinant protein from a precursor, preferably mammalian insulins, including human insulins and their analogues, characterised in that a protease is used which hydrolyses one or more peptide bonds in this protein, wherein the protease disrupts the peptide bond from the C-end side of a basic amino acid, when the amino acid is the second one after other basic or neutral amino acid, and such an order enables specific recognition of both amino acids by the protease. The invention is applicable in biotechnology and pharmaceutical industry. 122-. (canceled)23. A method for preparation of insulin or insulin analogue from a recombinant protein precursor , comprising wherein the protease hydrolyzes one or more peptide bonds in the precursor,', 'wherein the protease disrupts a peptide bond from a C-end side of a basic amino acid,', 'wherein such order of the amino acids enables specific recognition of both amino acids by the protease, and', 'wherein in the precursor the amino acid having the disrupted peptide bond is in an order such that it is a second one after another basic or neutral amino acid,'}], 'a) using a protease Kex2 in in vitro conditions,'}b) expressing in bacteria the resultant insulin or insulin analogue precursor.24Escherichia coli.. The method according to claim 23 , wherein in (b) the bacteria in which the insulin or insulin analogue precursor is expressed is25. The method according to claim 23 , and further comprising 'wherein the protein C-end is Kex1 or carboxypeptidase B in vitro.', 'wherein the exopeptidase hydrolyzes one or more peptide bonds of a basic amino acid at a protein C-end,'}, 'prior to (b) subjecting the protein obtained in (a) to treatment with a second protease which is an exopeptidase,'}26. The method according to claim 23 , wherein in (a) the amino acids are both neutral and basic amino acids.27. The method according to claim 24 , wherein in (a) the amino acids are both neutral and basic amino acids.28. ...

Fusion Protease

This invention relates to novel bifunctional fusion proteases useful for manufacturing a mature protein from a fusion protein. More specifically the present invention relates to bifunctional fusion proteases comprising a picornaviral 3C protease and a Xaa-Pro-dipeptidyl aminopeptidase. 1. A bifunctional fusion enzyme comprising the catalytic domains of a picornaviral 3C protease and a XaaProDAP.2. The bifunctional fusion protease according to comprising a protein of the formula:{'br': None, 'X—Y—Z\u2003\u2003(I) or'}{'br': None, 'Z—Y—X\u2003\u2003(II)'}whereinX is a picornaviral 3C protease or a functional variant thereof;Y is an optional linker;Z is a Xaa-Pro-dipeptidyl aminopeptidase (XaaProDAP) or a functional variant thereof;wherein said fusion protease has substantially no self-cleavage activity able to deteriorate at least one of the two proteolytic activities.3. The bifunctional fusion protease according to comprising a protein of formula (I) claim 2 , wherein said picornaviral 3C protease or a functional variant thereof is in the N-terminal part of said bifunctional fusion protease.4. The bifunctional fusion protease according to claim 2 , wherein X is a human Rhinovirus 3C protease or a functional variant thereof.5. The bifunctional fusion protease according to claim 2 , wherein X comprises SEQ ID NO: 2 claim 2 , or a functional variant thereof.6. The bifunctional fusion protease according to claim 2 , wherein Z is an E.C. 3.4.14.11 enzyme or a functional variant thereof.7. The bifunctional fusion protease according to claim 6 , wherein Z is an enzyme from a lactic acid bacterium or a functional variant thereof.8. The bifunctional fusion protease according to claim 2 , wherein Z is SEQ ID NO: 1 or a functional variant thereof.9Streptococcus. The bifunctional fusion protease according to claim 2 , wherein Z is an enzyme from spp. or a functional variant thereof.10. The bifunctional fusion protease according to wherein Z is SEQ ID NO: 24 or a functional ...

USE OF PROLINE SPECIFIC ENDOPROTEASES TO HYDROLYSE PEPTIDES AND PROTEINS

The present invention relates to a process for the proteolytic hydrolysis of a peptide or a polypeptide, said peptide or polypeptide comprising 4 to 40, preferably 5 to 35, amino acid residues and said peptide or polypeptide is not hydrolysable by subtilisin whereby said peptide or polypeptide is hydrolysed by a proline specific endo protease at a pH of 6.5 or lower, preferably 5.5 or lower and more preferably 5.0 or lower to hydrolyse said peptide or polypeptide. 119-. (canceled)20. A process for proteolytic hydrolysis of a peptide or a polypeptide ,wherein said peptide or polypeptide comprising 4 to 40, preferably 5 to 35, amino acid residues, andwherein said peptide or polypeptide is not hydrolysable by subtilisin,said process comprising the step of:hydrolyzing said peptide or polypeptide with a proline specific endo protease at a pH of 6.5 or lower, preferably 5.5 or lower and more preferably 5.0 or lower to hydrolyse said peptide or polypeptide.21. A process for the proteolytic hydrolysis of a peptide or a polypeptide ,wherein said peptide or polypeptide comprising 4 to 40, preferably 5 to 35 amino acid residues, andwherein said peptide or polypeptide comprises the tripeptide motif Glu-Xxx-Pro, Gln-Xxx-Pro, Tyr-Pro-Phe or Tyr-Pro-Trpsaid process comprising the step of:hydrolyzing said peptide or polypeptide by a proline specific endo protease at a pH of 6.5 or lower, preferably 5.5 or lower and more preferably 5.0 or lower to hydrolyse said peptide or polypeptide.22. A process for proteolytic hydrolysis of a peptide or a polypeptide ,wherein said peptide or polypeptide comprises 4 to 40, preferably 5 to 35 amino acid residues,wherein the amino acid residues of the peptide or polypeptide comprises for at least 30%, preferably at least 40%, proline and/or glutamine residues,said process comprising the step of:hydrolyzing said peptide or polypeptide by a proline specific endo protease at a pH of 6.5 or lower, preferably 5.5 or lower and more preferably 5.0 or ...

METHOD FOR PRODUCING A RECOMBINANT PROTEIN OF INTEREST

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 chaotropic conditions, (c) binding the fusion protein of the solubilised inclusion bodies to a multimodal chromatographic material under chaotropic conditions, (d) eluting the fusion protein from the multimodal chromatographic material with an elution buffer and 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 chaotropic conditions,(c) binding the fusion protein of the solubilised inclusion bodies to a multimodal chromatographic material under chaotropic conditions,{'sup': 'pro', '(d) eluting the fusion protein from the multimodal chromatographic material with an elution buffer and 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.2E. coli. Method according to claim 1 , characterized in that the inclusion bodies were generated in a recombinant production system claim 1 , preferably in a prokaryotic host cell claim 1 , especially in host cells.3. Method according to claim 1 , characterized in that the chaotropic conditions in step (b) correspond to a urea concentration of ...

ANTI-VEGF PROTEIN COMPOSITIONS AND METHODS FOR PRODUCING THE SAME

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions. 1. A composition comprising oxo-aflibercept , wherein one or more amino acid residues of aflibercept is oxidized and wherein said one or more amino acid residues is histidine and/or tryptophan.2. (canceled)3. The composition of claim 1 , wherein said oxo-aflibercept is enzymatically digested resulting in one or more oligopeptides claim 1 , and wherein said one or more oligopeptides is selected from the group consisting of: SEQ ID NO. 17 claim 1 , SEQ ID NO. 18 claim 1 , SEQ ID NO. 19 claim 1 , SEQ ID NO. 20 claim 1 , SEQ ID NO. 21 claim 1 , SEQ ID NO. 22 claim 1 , SEQ ID NO. 23 claim 1 , SEQ ID NO. 28 claim 1 , SEQ ID NO. 29 claim 1 , SEQ ID NO. 30 claim 1 , SEQ ID NO. 31 claim 1 , SEQ ID NO. 32 and combinations thereof.4. The composition of claim 3 , wherein said enzymatic digestion is performed using trypsin.5. A method of producing oxidized species of aflibercept comprising subjecting a sample having aflibercept to cool-white light from about 0.24 million lux*hr to about 2.4 million lux*hr.6. The method of claim 5 , wherein said aflibercept is subjected to cool-white light for about 0.24 million lux*hr and wherein the amount of oxidized species of aflibercept increases by about 1.5 to about 10-fold compared to untreated aflibercept.7. The method of claim 5 , wherein said aflibercept is subjected to cool-white light for about 0.96 million lux*hr and wherein the amount of oxidized species of aflibercept increases by about 1.5 to about 20-fold compared to untreated aflibercept.8. The method of claim 5 , wherein said aflibercept is subjected to cool-white light for about 1.2 million lux*hrs and wherein the amount of oxidized species of aflibercept increases by about 1.5 to about 20-fold compared to untreated aflibercept.9. The method of claim 5 , wherein said aflibercept is subjected to cool-white light for about 2.4 million lux*hr wherein the ...

PROTEIN RETROSPLICING ENABLED BY A DOUBLE LIGATION REACTION

Proteins containing a C-terminal thioester are important intermediates in semi-synthesis. Currently there is one main method for the synthesis of protein thioesters that relies upon the use of engineered inteins. The invention involves, in some aspects a method, utilizing Sortase A, for preparation of recombinant proteins containing a C-terminal thioester. This new method for double ligatation is useful for synthesizing new or naturally occurring molecules such as a protein thioester. 16-. (canceled)7. A method comprisingperforming a ligation reaction of a N-terminal protein domain with a peptide thioester in the presence of a cysteine transpeptidase enzyme to produce a N-terminal protein domain-COSR product,reacting the N-terminal protein domain-COSR product with a C-terminal protein domain, wherein the C-terminal protein domain has a cysteine at the N-termini, to produce a modified protein having a chemical entity linking the N-terminal protein domain and the C-terminal protein domain.8. The method of claim 7 , wherein the cysteine transpeptidase enzyme is a SrtA enzyme.9. The method of claim 8 , wherein SrtA is SrtA*.10. The method of claim 7 , wherein the ligation reaction is performed in a sortase buffer.11. The method of claim 7 , whereinprior to the ligation reaction the N-terminal protein or N-terminal protein domain is recombinantly expressed as a SUMO-protein having a SUMO tag.12. The method of claim 11 , wherein the SUMO tag is removed using SUMO protease.13. The method of claim 7 , wherein the peptide thioester is G-Xaa-COSR (SEQ ID NOs:1-3) claim 7 , wherein n is 1-6 and wherein Xaa is an amino acid.14. The method of claim 7 , wherein the peptide thioester is G-Xaa-COSR (SEQ ID NOs:4-5) claim 7 , wherein n is 3-5 and claim 7 , wherein Xaa is Gly claim 7 , Phe claim 7 , Ser or Leu.15. The method of claim 7 , wherein the peptide thioester is GGGGG-Xaa-COSR (SEQ ID NOs:5) claim 7 , wherein Xaa is Gly claim 7 , Phe claim 7 , Ser or Leu.16. The method of ...

METHOD

The present invention relates to a method of producing keratin hydrolysate comprising the steps of: i) reacting keratin material with a protease; and ii) reacting keratin material with a chemical oxidant; wherein step ii occurs: a) after step i); b) during step i) when the selected protease hydrolyses under the pH conditions used for the chemical reaction and/or c) prior to step i) when the selected protease hydrolyses under the reaction conditions used for the chemical reaction; keratin hydrolysate so produced and uses thereof. 3. A method according to or , wherein the selected protease hydrolyses under the pH conditions used for the chemical reaction.4Bacillus.. A method according to any one of the preceding claims wherein the protease is from the genus5. A method according to any one of the preceding claims wherein the protease comprises a polypeptide sequence as defined in any one of SEQ ID NOs: 1 or 2 , or a functional fragment or variant thereof , having at least 75% sequence Identity to any one of SEQ ID NOs: 1 or 2 over at least 50 amino acid residues.6. A method according to any one to wherein the protease may be transcribed from a nucleic acid sequence encoding a protease with at least 75% identity to either SEQ ID NO3 or SEQ ID N04 or a nucleic acid sequence capable of hybridizing to the nucleic acid sequence of SEQ ID NO3 or SEQ ID NO4 or the complement thereof under stringent conditions.7. A method according to any one of the preceding claims , wherein the chemical is an oxidant.8. A method according to any one of the preceding claims , wherein the chemical is one or more of the chemicals selected from the group consisting of: sodium chlorite , HCl , acetic acid , hydroxyacetic acid , NaOH , peracids , HOCl , HOBr , NaClO , ClO , HO , ammonium hydroxide , sodium hydroxide , and calcium hydroxide.9. A method according to any one of the preceding claims , wherein the chemical is added prior to and/or during step i) and the chemical oxidant adjusts the pH ...

Mutant Proteinase with Reduced Self-Cleavage Activity and Method of Purification

The present invention provides a mutant 27 kDa NIa proteinase having reduced self-cleavage activity relative to the self-cleavage activity of its wild-type proteinase. The mutant has the same substrate cleavage activity as the wild-type proteinase but is more stable than the wild-type proteinase. The present invention also provides a method of obtaining large quantities of active 27 kDa NIa proteinase for use as a tool for purification of other proteins. 1. An isolated mutant proteinase having a molecular weight of about 27 kDa and reduced self-cleavage activity relative to the self-cleavage activity of its wild-type proteinase.2. The mutant proteinase of claim 1 , wherein the mutant proteinase comprises an amino acid sequence in which the residue corresponding to Ser 219 of the 27 kDa NIa proteinase is replaced with another residue.3. The mutant proteinase of claim 2 , wherein the residue replacing Ser 219 of 27 kDa NIa proteinase is Asn.4. A composition comprising the mutant proteinase of and a carrier.5. A fusion protein comprising the mutant proteinase of fused to a heterologous polypeptide.6. The fusion protein of claim 5 , wherein the heterologous polypeptide consists of six histidines.7. An isolated nucleic acid molecule comprising a sequence encoding a mutant proteinase having a molecular weight of about 27 kDa and reduced self-cleavage activity relative to the self-cleavage activity of its wild-type proteinase.8. An isolated nucleic acid molecule of claim 7 , wherein the nucleic acid molecule encodes an amino acid sequence in which the residue corresponding to Ser 219 of the 27 kDa NIa proteinase is replaced with another amino acid.9. An isolated nucleic acid of claim 8 , wherein the nucleic acid comprises a sequence that encodes an amino acid sequence in which the residue corresponding to Ser 219 of the 27 kDa NIa proteinase in Asn.10. A vector comprising the nucleic acid of .11. An expression vector comprising the nucleic acid of .12. A host cell ...

Compositions and Methods of Use for Recombinant Human Secretoglobins

Methods of synthetically producing, formulating and using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 are provided. Methods of using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 as therapeutic agents to affect long term patient outcomes, such as preventing severe respiratory exacerbations of underlying conditions that require medical intervention, including hospitalization are provided. Methods of producing recombinant human secretoglobins, analytical methods, pharmaceutical compositions, and methods of use to prevent the long term sequelae of acute and chronic respiratory conditions are provided. 112-. (canceled)13. A composition of matter comprising recombinant human SCGB3A2 protein having an N-terminus ATA , namely SEQ ID 1.14. The protein of claim 13 , wherein the protein inhibits PLA2 enzymes.15. The protein of claim 13 , wherein the protein migrates in an isoelectric focusing gel corresponding to an isoelectric point at or between 6.3-6.7.16. The protein of wherein the protein further comprises a homodimer.17. The protein of wherein the protein further comprises a homodimer having a pI of 6.7.18. A process for synthesizing recombinant human SCGB3A2 comprising: using a UBL fusion protein and UBL protease that recognizes the fusion partner and cleaves between the fusion partner and SCGB3A2 claim 14 , to release an intact SCGB3A2 protein claim 14 , namely SEQ ID 1. This application is a divisional of U.S. patent application Ser. No. 13/843,289, filed Mar. 15, 2013, now pending, the disclosures of which are hereby incorporated by reference in their entirety.The invention relates to pharmaceutical compositions, methods of production, analytical methods, and methods of use for secretoglobin proteins, including SCGB1A1 (CC10), SCGB3A1, and SCGB3A2. Novel physiologic roles and therapeutic uses for these secretoglobins have been identified. Specifically, the present invention relates to novel methods of use for rhCC10, rhSCGB3A2, and rhSCGB3A1 in preventing or ...

CHONDROCYTE PROLIFERATION PROMOTING AGENT

The present invention provides a novel active ingredient which can be safely used in preventing, ameliorating or treating diseases related to the cartilage such as cartilage damages and cartilage disorders. Since the egg yolk protein hydrolyzate has an effect on chondrocyte proliferation promotion, it is useful as an active ingredient for preventing, ameliorating or treating cartilage disorders and for preventing, ameliorating or treating joint pain. The egg yolk protein hydrolyzate is a natural-origin material with high safety and, therefore, can be widely used in foods and drinks, medicines, feeds and the like which can be daily ingested. 17-. (canceled)8. A method of preventing or ameliorating cartilage disorder or joint pain characterized by orally administering the egg yolk protein hydrolyzate to human in need of promoting chondrocyte proliferation.9. A manufacturing method of chondrocyte proliferation promoting agent characterized by comprising a step of defatting egg yolk , a step of obtaining the egg yolk protein hydrolyzate by hydrolyzing the resulting defatted egg yolk with a proteolytic enzyme , and a step of comprising the resulting egg yolk protein hydrolyzate as an active ingredient.10. A method of preventing , treating or ameliorating cartilage disorder or joint pain characterized by administering the egg yolk protein hydrolyzate to animal.11. A method of promoting chondrocyte proliferation characterized by administering the egg yolk protein hydrolyzate to animal. This invention relates to chondrocyte proliferation promoting agent, preventing or ameliorating agent for cartilage disorders, and preventing or ameliorating agent for diseases related to the cartilage of joint pain and the like, and is particularly directed to chondrocyte proliferation promoting agent, preventing or ameliorating agent for cartilage disorders, and preventing or ameliorating agent for diseases related to the cartilage such as joint pain including the egg yolk protein ...

PROTEIN DEGRADATION INDUCING TAG AND USAGE THEREOF

Provided are: a protein degradation inducing tag which is a molecule that has affinity with proteases and does not inhibit degradation of a protein by proteases; a protein degradation inducing molecule that is a conjugate of at least one protein degradation inducing tag and at least one protein binding molecule that binds to a protein; and a usage of those. 1. A protein-degradation inducing tag , which is a molecule having a molecular weight of 5000 or less and having an affinity with a protease without inhibiting degradation of a protein by the protease.2. (canceled)3. The protein-degradation inducing tag according to claim 1 , wherein the protease is a proteasome.4. (canceled)5. (canceled)6. A method of screening for a protein-degradation inducing tag claim 1 , the method including a step of selecting a molecule having an affinity with a protease without inhibiting degradation of a protein by the protease from candidate molecules having a molecular weight of 5000 or less.7. The method of screening for a protein-degradation inducing tag according to claim 6 , wherein the protease is a proteasome.8. A method of manufacturing a protein-degradation inducing tag claim 6 , the method including a step of modifying a structure of an active site of a protease inhibitor to inactivate a protease inhibitory activity.9. The method of manufacturing a protein-degradation inducing tag according to claim 8 , the method further including a step of selecting claim 8 , as the protease inhibitor claim 8 , a molecule having an affinity with a protease and inhibiting degradation of a protein by the protease from candidate molecules.1022.-. (canceled) The present disclosure relates to a protein-degradation inducing tag and use thereof.Controlling the amount (expression) of a target protein in a cell, a living body, and the like is very useful for analyzing the functions of the target protein and life phenomena in which the target protein is involved. When a target protein is responsible ...

PRODUCTION OF ACTIVATED CLOSTRIDIAL NEUROTOXINS

The present invention relates to a method of producing activated clostridial neurotoxins that are essentially free of unactivated products, to compositions comprising such and to their use in therapy. 1. Method for producing an activated clostridial neurotoxin , comprising contacting a single chain clostridial neurotoxin with an activation enzyme until at least 90% of the single chain clostridial neurotoxin is converted into a di-chain clostridial neurotoxin.2. Method according to claim 1 , wherein said activation enzyme is a trypsin.3. Method according to claim 2 , wherein said trypsin is a bovine trypsin and wherein said bovine trypsin has an amino acid sequence which has at least 90% identity to SEQ ID NO: 1.4. Method according to claim 3 , wherein said bovine trypsin is selected from a native trypsin obtained from bovine pancreas and a recombinant bovine trypsin.5. Method according to or claim 3 , wherein said step of contacting said single chain clostridial neurotoxin with a bovine trypsin is performed at a pH of between 5 and 7 claim 3 ,5 claim 3 , preferably between 6 and 7 claim 3 , for example at a pH of approximatively 6 claim 3 ,5.6. Method according to claim 5 , wherein the step of contacting the single chain clostridial neurotoxin with said bovine trypsin is carried out at room temperature at a pH between 6 and 7 for a duration of 15 to 25 hours claim 5 , and wherein the concentration of said bovine trypsin is between 0.5 and 3 μg per mg of clostridial neurotoxin.7. Method according to any one of to claim 5 , further comprising a step of removing truncated clostridial neurotoxin.8. Method according to claim 7 , wherein said step of removing truncated clostridial neurotoxin comprises contacting said activated clostridial neurotoxin with a mixed mode chromatography resin.9. Method according to any one of to claim 7 , wherein said clostridial neurotoxin is selected from BoNT/A claim 7 , BoNT/B claim 7 , BoNT/C claim 7 , BoNT/D claim 7 , BoNT/E claim 7 , ...

Collagen Peptide Composition Production Method, DPP-4 Inhibitor, and Antihyperglycemic Agent

Provided are methods of producing a novel collagen peptide composition, and a DPP-4 inhibitor and antihyperglycemics that comprise the above-mentioned collagen peptide composition. Ginger rhizome-derived enzymes are added to and break down a collagen and/or gelatin solution to generate peptide compositions comprising peptides represented by X-Hyp-Gly (wherein X represents an amino acid residue other than Gly, Hyp, and Pro). The thus obtained collagen peptide composition has a high DPP-4 inhibitory activity and an excellent antihyperglycemic effect. 1. A method of producing a collagen peptide composition comprising a peptide represented by X-Hyp-Gly (wherein X represents an amino acid residue other than Gly , Hyp , and Pro) by degrading a collagen and/or gelatin with the addition of a ginger rhizome-derived enzyme into the collagen and/or gelatin solution to generate the peptide represented by the formula.2. The method of producing a collagen peptide composition according to claim 1 , wherein a dried ground product of ginger rhizome is used as the ginger rhizome-derived enzyme.3. The method of producing a collagen peptide composition according to claim 1 , wherein further a glutathione-containing yeast extract in a range of 0.005 to 0.5 w/v % is added with the ginger rhizome-derived enzyme claim 1 , and/or a pH is adjusted in a range of 4.0 to 6.0.4. The method of producing a collagen peptide composition according to claim 1 , wherein a content of the peptide represented by the formula in the peptide composition is not less than 0.01% by mole of the collagen and/or gelatin.5. The method of producing a collagen peptide composition according to claim 1 , wherein the peptide composition further comprises a peptide represented by X-Pro-Gly (wherein X represents an amino acid residue other than Gly claim 1 , Hyp claim 1 , and Pro).6. A DPP-4 inhibitor comprising the collagen peptide composition obtained by the method of production according to .7. An agent for inhibiting ...

COLLAGEN MIXTURE AND METHOD OF MAKING THE SAME

A collagen mixture having a portion of unhydrolyzed eggshell membrane collagen and Avian collagen. 1. A method of making a collagen mixture , comprising the steps of:obtaining a portion of unhydrolyzed eggshell membrane collagen in dry powder form and a portion of avian collagen; andcombining the portion of unhydrolyzed eggshell membrane collagen and the avian collagen to form a combined portion that includes type III collagen.2. The method of wherein the combined portion includes type I claim 1 , II claim 1 , IV claim 1 , V claim 1 , and X collagen claim 1 , and lysozyme claim 1 , ovatransferin claim 1 , and sialic acid (LOS).3. The method of wherein the combined portion comprises a minimum of 15 percent mucopolysaccharide claim 1 , a minimum of 5 percent chondroitin sulfate claim 1 , and a minimum of 2 percent hyaluronic acid.4. The method of wherein the combined portion comprises a minimum of 20 percent protein.5. The method of wherein the combined portion comprises a minimum of 2 percent mucopolysaccharide claim 1 , a minimum of 0.1 percent chondroitin sulfate claim 1 , and a minimum of 0.5 percent hyaluronic acid.6. The method of wherein the combined portion comprises a minimum of 2 percent protein.7. The method of wherein the combined portion comprises a minimum of 0.3 percent glucosamine.8. The method of further comprising the step of adding an enzyme to avian cartilage to produce the portion of avian collagen.9. The method of wherein the enzyme includes a liquid plant derived enzyme.10. The method of wherein the enzyme includes a plant derived proteolytic enzyme.11. The method of wherein the enzyme includes a plant derived proteolytic enzyme claim 8 , a bacterial enzyme claim 8 , and a buffer. This application is a continuation of U.S. patent application Ser. No. 13/848,389 filed Mar. 21, 2013, which is a continuation-in-part of U.S. patent application Ser. No. 13/562,680 filed Jul. 31, 2012 which is a division of U.S. patent application Ser. No. 13/197,318 ...

MODIFIED CLEAVASES, USES THEREOF AND RELATED KITS

Provided herein are modified cleavases for removing amino acids from peptides, polypeptides, and proteins. Also provided are methods of using the modified cleavases for treating polypeptides, and kits comprising the modified cleavase. In some embodiments, the methods and the kits also include other components for macromolecule sequencing and/or analysis. 1. A modified cleavase comprising an unmodified cleavase comprising a mutation in a substrate binding site , wherein:the unmodified cleavase is a dipeptide cleavase and said modified cleavase removes or is configured to remove a single labeled terminal amino acid from a polypeptide; orthe unmodified cleavase is a tripeptide cleavase and said modified cleavase removes or is configured to remove a single labeled terminal amino acid from a polypeptide or a single labeled terminal dipeptide from a polypeptide.2. The modified cleavase of claim 1 , wherein the modified cleavase derived from the dipeptide cleavase or tripeptide cleavase is configured to cleave a peptide bond between a terminal labeled amino acid residue and a penultimate terminal amino acid residue of the polypeptide.34-. (canceled)5. The modified cleavase of claim 2 , wherein the modified cleavase derived from the dipeptide cleavase does not remove an unlabeled terminal dipeptide from the polypeptide.6. The modified cleavase of claim 1 , wherein the unmodified cleavase is a dipeptidyl peptidase 3 claim 1 , dipeptidyl peptidase 5 claim 1 , dipeptidyl peptidase 7 claim 1 , dipeptidyl peptidase 11 claim 1 , dipeptidyl aminopeptidase BII claim 1 , dipeptidyl peptidase BII claim 1 , or a protein classified in EC 3.4.14 claim 1 , EC 3.4.15 claim 1 , MEROPS S8 claim 1 , MEROPS S9 claim 1 , MEROPS S33 claim 1 , MEROPS S46 claim 1 , MEROPS M49 claim 1 , or MEROPS S53 claim 1 , or a functional homolog or fragment thereof.7. The modified cleavase of claim 2 , wherein the single labeled terminal amino acid is an N-terminal labeled amino acid of the polypeptide claim ...

Process for Producing Protein Concentrate or Isolate and Cellulosic Thermochemical Feedstock From Distillers Grains

A process for treating distillers grains to produce a high value protein product and a cellulosic residue both from distillers grains. The high value protein product is useful as a protein supplement or feed for livestock and poultry and the cellulosic residue has value as a feedstock for a thermochemical process unit for the production of a biofuel. 1. A process for producing a protein product and a cellulosic product suitable as a feedstock for thermochemical processing from distillers grains containing a protein component , which process comprises:a) milling said distillers grains to an average particle size of less than or equal to 0.5 mm.b) mixing said milled distillers grains with an effective amount of water to provide a an aqueous mixture wherein the water to grain weight ratio is about 10:1;c) adjusting the pH of the aqueous mixture with an aqueous basic solution to a pH of about 10.5;d) heating said pH adjusted aqueous mixture to a temperature of about 50° C. and keeping it at that temperature for about one hour;e) adjusting the pH of the heated aqueous mixture with an aqueous acid solution to a pH of about 9;f) adding an effective amount of a protease enzyme at a dose of about 10 mls/kg to the heated pH adjusted aqueous mixture of step e) above;g) maintaining the pH of the mixture of step f) above until the degree of hydrolysis of proteins of about 5 is reached, thereby resulting in a liquid fraction containing hydrolyzed proteins, and a solids fraction comprised of protein-lean cellulosic distillers grains;h) separating said liquid fraction from said solids fraction;i) spray drying said liquid fraction resulting in spray dried hydrolyzed protein product; andj) drying said solids fraction and collecting the protein-lean cellulosic distillers grains.2. The process of wherein the distillers grains are wet and are pretreated with ultrasonic energy.3. The process of wherein the distillers grains are a by-product from a corn to ethanol process.4. The process ...

Method for High-Yield Fermentation of Recombinant Proline Aminopeptidase and Preparation of Debittered Rice Peptide

The present disclosure discloses methods for high-yield fermentation of recombinant proline aminopeptidase and preparation of debittered rice peptide, belonging to the fields of fermentation technology, enzyme preparation and food additives. The present disclosure utilizes fermentation kinetic analysis to determine the high-yield fermentation method of proline aminopeptidase by recombinant and improve the yield of proline aminopeptidase to reach 174.8 U/mL. Proline aminopeptidase cooperates with alkaline protease and leucine aminopeptidase to hydrolyze rice protein. The free amino acid content is 27.3 times the unhydrolyzed free amino acid content, and the small peptide content below 180 Da in hydrolysate reaches 44.70%. The exposed N-terminal proline residue is fully hydrolyzed, and the free proline content is 1,064.3 times that of the unhydrolyzed free proline content, which increases the degree of rice protein hydrolysis. The method of the present disclosure has a good application prospect in the fields of foods and beverages and processing and utilization of food protein resources. 1Bacillus subtilis,Bacillus subtilis. A method for high-yield fermentation production of a proline aminopeptidase using a recombinant wherein the recombinant comprises a gene encoding the proline aminopeptidase , comprising:{'i': 'Bacillus subtilis', '(1) inoculating a seed medium with the recombinant and culturing to prepare a seed solution;'}(2) inoculating a fermentation medium with the seed solution, introducing sterile air at an aeration rate of 1.2 to 1.5 vvm, wherein rotational speed is adjusted as: 200 rpm from 0 to 6 h, 400 rpm from 6 to 12 h, 500 rpm from 12 to 28 h, and 400 rpm from 28 h to the end of fermentation; wherein adjustment of pH is: applying no pH control from 0 to 12 h, setting pH as 7.0 from 12 to 16 h, applying no pH control from 16 to 28 h, and setting pH as 7.0 after 28 h; and wherein temperature is adjusted as: 40° C. from the beginning of fermentation to 8 ...

METHOD FOR THE DEGRADATION OF KERATIN AND USE OF THE KERATIN HYDROLYSATE PRODUCED

The present invention relate to a method for producing keratin hydrolysate comprising the step of: i) admixing keratin material with a protease and a non-sulphur containing surfactant, as well as keratin hydrolysate so produced and uses thereof. 1. A method of producing keratin hydrolysate comprising the step of:i) admixing keratin material with a protease and a non-sulphur containing surfactant.2. A method according to wherein step i) occurs under controlled oxygen levels.3. A method of degrading keratin wherein a keratin material is admixed with a protease and a non-sulphur containing surfactant.4. A method according to any one of to claim 1 , wherein the surfactant is selected from one or more of the group consisting of: sodium decanoate; Triton-X-100; Tween 20; Tween 80; lecithin; polyoxyethylene stearate; polyoxyethylene sorbitan monolaurate; polyoxyethylene sorbitan monooleate; polyoxyethylene sorbitan monopalmitate; polyoxyethylene sorbitan monostearate; polyoxyethylene sorbitan tristearate; ammonium phosphatides; sodium claim 1 , potassium or calcium salts of fatty acids; magnesium salts of fatty acids; acetic acid esters of mono- and diglycerides of fatty acids; lactic acid esters of mono- and diglycerides of fatty acids; citric acid esters of mono- and diglycerides of fatty acids; mono- and diacetyl tartaric acid esters of mono- and diglycerides of fatty acids; sucrose esters of fatty acids; sucroglycerides; polyglycerol esters of fatty acids; polyglycerol polyricinoleate; propane-1 claim 1 ,2-diol esters of fatty acids; thermally oxidised soya bean oil interacted with mono- and diglycerides of fatty acids; sodium stearoyl-2-lactylate; calcium stearoyl-2-lactylate; sorbitan monostearate; sorbitan tristearate; sorbitan monolaurate; sorbitan monooleate and sorbitan monopalmitate.5. A method according to any one of to claim 1 , wherein the surfactant is sodium stearoyl-2-lactylate and/or diacetyltartaric acid esters of mono- and diglycerides.6. A method ...

PEPTIDES FROM FISH GELATINE

The invention relates to a process of preparing low molecular weight peptides from gelatine. The gelatine is broken down into low molecular weight peptides using enzymes from sp. to obtain peptides with low ash content. The gelatine is clarified by adjusting the pH to 7 using alkali hydroxide and filtered. The filtered solution is desalted at 20 to 55° C. in a diafiltration mode till the salt content decreases below 0.05% (w/v) The product so obtained is hydrolysed with an enzyme maintaining pH in the range of 7 to 10 and at temperature in the range of 25 to 75° C. The hydrolysed product is filtered through ultrafiltration membrane at temperature in the range of 15 to 55° C. and permeate is concentrateds at temperature in the range of 50 to 100° C. under vacuum to obtain the peptide solution of the invention. 1. A process for preparing low molecular weight peptide comprising the steps of:a. clarifying 0.5 to 6.5% w/v fish gelatine solution by adjusting pH up to 7 followed by centrifuging the solution followed by membrane filtration to make the solution particle-free by discarding the solid particles;b. desalting the solution as clarified in step (a) through ultrafiltration membrane at temperature in the range of 20 to 55° C. in a diafiltration mode till the salt content decreases below 0.05% (w/v) followed by discarding the permeate;c. hydrolyzing the product obtained in step (b) with an enzyme maintaining pH in the range of 7 to 10 and at temperature in the range of 25 to 75° C. for period in the range of 3.5 to 4 hr to obtain hydrolysed product;d. filtering the hydrolyzed product of step (c) through ultrafiltration membrane at temperature in the range of 15 to 55° C. and concentrating the permeate at temperature in the range of 50 to 100° C. under vacuum followed by drying by any conventional means known in the art such as spray drying to obtain peptide powder.2. The process as claimed in claim 1 , wherein said peptide powder exhibit molecular weight less than ...

SPLIT INTEINS AND USES THEREOF

The present invention relates generally to robust split inteins. The split inteins described herein are active over a large temperature range, including temperatures as low as 0° C., over a wide pH range, and in the presence of chaotropic salts. The split inteins also show high tolerance to sequence variability in fused heterologous polypeptides and therefore are useful in protein purification and engineering techniques. 115.-. (canceled)16. A fusion protein comprising (i) an intein domain at least 75% identical to a sequence selected from the group consisting of SEQ ID NOs: 7 , 16 , 24 , 38 and 65 and (ii) a heterologous polypeptide , wherein the heterologous polypeptide is C-terminal to the intein domain.17. The fusion protein of wherein the last amino acid of the intein domain is glutamine or asparagine.18. The fusion protein of wherein the first amino acid of the heterologous polypeptide is serine claim 17 , cysteine or threonine.19. The fusion protein of wherein the last amino acid of the intein domain is an amino acid other than asparagine or glutamine and wherein the first amino acid of the heterologous polypeptide is an amino acid other than serine claim 16 , cysteine claim 16 , or threonine.20. A fusion protein comprising (i) an intein domain at least 75% identical to a sequence selected from the group consisting of SEQ ID NOs: 3 claim 16 , 12 claim 16 , 20 claim 16 , 34 and 64 and (ii) a heterologous polypeptide claim 16 , wherein the heterologous polypeptide is N-terminal to the intein domain.21. The fusion protein of claim 20 , wherein the first amino acid of the intein domain is a serine or cysteine.22. The fusion protein of wherein the first amino acid of the intein domain is an amino acid other than serine or cysteine.23. A composition or kit-of-parts comprising a first component and a second component wherein{'claim-ref': {'@idref': 'CLM-00016', 'claim 16'}, '(i) the first component is the fusion protein of and'}(ii) the second component is selected ...

METHODS OF ACTIVATING CLOSTRIDIAL TOXINS

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 method of activating a modified Clostridial toxin , the method comprising the step of incubating a modified Clostridial toxin with a BoNT/A di-chain loop protease under physiological conditions;wherein the BoNT/A toxin di-chain loop protease is selected from the group consisting of SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37 and SEQ ID NO: 38;wherein the modified Clostridial toxin comprises an exogenous BoNT/A di-chain loop protease cleavage site and is selected from the group consisting of a modified BoNT/B, a modified BoNT/C, a modified BoNT/D, a modified BoNT/E, a modified BoNT/F, a modified BoNT/G, a modified TeNT, a modified BaNT, a modified BuNT, and a Clostridial toxin chimeric variant; andwherein cleavage of the modified Clostridial toxin by the BoNT/A di-chain loop protease converts the modified Clostridial toxin from its single-chain polypeptide form into its di-chain form, thereby activating the modified Clostridial toxin.2. The method of claim 1 , wherein the modified Clostridial toxin is a modified BoNT/B.3. The method of claim 1 , wherein the modified Clostridial toxin is a modified BoNT/C.4. The method of claim 1 , wherein the modified Clostridial toxin is a modified BoNT/E.5. The method of claim 1 , wherein the modified Clostridial toxin chimeric variant.6. A method of activating a recombinantly expressed Clostridial toxin claim 1 , the method comprising the steps of:(a) expressing in a cell a polynucleotide molecule encoding a modified Clostridial toxin, wherein the modified Clostridial toxin comprises an ...

THIOL-BASED DEEP EUTECTIC SOLVENT

A deep eutectic solvent consisting of (2-hydroxyethyl) trimethyl ammonium chloride and dithiothreitol in a molar ratio of from 1:2 to 1:3 and from 0% to 10% co-solvent, and methods of enzymatic production of polypeptides using the deep eutectic solvent. 1. A deep eutectic solvent consisting of (2-hydroxyethyl) trimethyl ammonium chloride and dithiothreitol in a molar ratio of from 1:2 to 1:3 and from more than 0% to 10% of co-solvent.2. A deep eutectic solvent consisting ofa) i) (2-hydroxyethyl) trimethyl ammonium chloride, andii) 4-mercapto-1-butanol or 1-mercapto-2-propanol in a molar ratio of i) to ii) of about 1:2, andb) of from more than 0% to about 10% co-solvent.3. A deep eutectic solvent consisting of (2-hydroxyethyl) trimethyl ammonium chloride and sodium 2-mercaptoethansolfonate in a molar ratio of about 1:1 and of from more than 0% to about 10% co-solvent.4. The deep eutectic solvent of claim 1 , wherein the co-solvent is an aqueous co-solvent.5. The deep eutectic solvent of claim 4 , comprising from more than 0% up to about 5% (v/v) aqueous co-solvent.6. A method for the enzymatic production of a polypeptide comprising the following step i) a first polypeptide comprising the amino acid sequence LPXTG (SEQ ID NO: 01, wherein X can be any amino acid residue) or LPXTA (SEQ ID NO: 101, wherein X can be any amino acid residue),', 'ii) a second polypeptide comprising i) a glycinyl, an alaninyl, or a cysteinyl compound at its N-terminus, or ii) an oligoglycine, or oligoalanine, or a cysteine amino acid residue followed by one to three glycine or alanine amino acid residues at its N-terminus, or iii) a lysine amino acid residue within its 5 N-terminal amino acid residues, and', 'iii) a third polypeptide with sortase A activity,, 'incubating'}{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'in the deep eutectic solvent of ;'}wherein the polypeptide is produced.7. The method of claim 6 , wherein the second polypeptide has at its N-terminus the amino acid ...

INTERLEUKIN-6, 10 PRODUCTION PROMOTER

[Problem] To provide a novel technology that can promote the production of interleukin-6, and to provide a novel technology that can promote the production of interleukin-10. 14-. (canceled)5. A method for promoting interleukin-6 production in a subject , which comprises: administering a crude yeast cell wall hydrolysate to the subject.6. The method according to claim 5 , wherein the yeast cell wall hydrolysate is a yeast cell wall hydrolysate obtained by adjusting pH of yeast cell walls to 8.0 to 14.0 and hydrolyzing the yeast cell walls at 60 to 120° C. for 3 to 24 hours.7. A method for promoting interleukin-10 production in a subject claim 5 , which comprises: administering a crude yeast cell wall hydrolysate to the subject.8. The method according to claim 7 , wherein the yeast cell wall hydrolysate is a yeast cell wall hydrolysate obtained by adjusting pH of yeast cell walls to 8.0 to 14.0 and hydrolyzing the yeast cell walls at 60 to 120° C. for 3 to 24 hours. The present invention relates to interleukin-6, 10 production.Attacks of the immune system against pathogens that have invaded the body include phagocytosis by phagocytes such as neutrophils and macrophages (innate immune system), the disruption of host cells by the release of cytotoxic substances such as perforin from cytotoxic T cells, and the inactivation of pathogens by antibodies produced by B cells (adaptive immune system).Cytokines play important roles in the activation and the functional inhibition of cells involved in the immune system and such cytokines include interleukins secreted by leukocytes.A plurality of interleukins has been identified up to now. Among them, interleukin-6 (IL-6) is known to have actions such as induction of acute reactions by stimulating macrophages. Interleukin-10 (IL-10) is known to be mainly produced by type 2 helper T cells (Th2) and act on inhibition of inflammatory reactions.Meanwhile, the immunopotentiating effect of β-glucan, obtained by centrifugal purification ...

REVERSIBLE CHEMOENZYMATIC LABELING OF NATIVE AND FUSION CARRIER PROTEIN MOTIFS

Provided herein, inter alia, are methods and compositions for removing a phosphopantethiene analogue moiety from an ACP-phosphopantetheine conjugate thereby providing Apo-ACP proteins. 2. The method of claim 1 , wherein said ACP is an ACP protein fusion moiety.3. The method of claim 2 , wherein said ACP protein fusion moiety comprises an ACP protein moiety bound to an amino terminus or a carboxy terminus of a second fusion protein moiety.4. The method of claim 2 , wherein said ACP protein fusion moiety comprises an ACP protein moiety bound to an internal amino acid residue of a second fusion protein moiety.5P. aeruginosaCyanotheceP. fluorescens. The method of claim 1 , wherein said ACP hydrolase is a ACP hydrolase claim 1 , sp. ACP hydrolase claim 1 , or ACP hydrolase.6. The method of claim 1 , wherein said apo-ACP is a Fatty Acid apo-ACP claim 1 , Polyketide apo-ACP claim 1 , or Peptide apo-ACP.7E. coliP. aeruginosaS. oneidensisP. falciparumM. tuberculosisS. coelicolorA. parasiticusG. fujikuroiL. majusculeP. fluorescens. The method of claim 6 , wherein said apo-ACP is a apo-ACP claim 6 , apo-ACP claim 6 , apo-ACP claim 6 , apo-ACP claim 6 , apo-ACP claim 6 , apo-ACP claim 6 , apo-ACP claim 6 , apo-ACP claim 6 , apo-ACP or apo-ACP.9. The method of claim 8 , further comprising(i) contacting said second ACP-phosphopantetheine conjugate with a second ACP hydrolase; and(ii) allowing said second ACP hydrolase to cleave said phosphodiester linker thereby forming said Apo-ACP.10B. subtilis. The method of claim 8 , wherein said phosphopantetheinyl transferase is a phosphopantetheinyl transferase.11. A compound comprising an amino acid sequence having the formula:{'br': None, '-DSL(Aaa1)(Aaa2)(Aaa3)(Aaa4)(Aaa5)(Aaa6)-\u2003\u2003(I),'}wherein;Aaa1 is D, E, or S;Aaa2 is T, F, or W,Aaa3 is V, L, or I;Aaa4 is E, A, or L;Aaa5 is A, S, R, or L;Aaa6 is V, K, or L; andwherein said sequence is not -DSLDTVELV- (SEQ ID NO:97).14. The compound of claim 13 , wherein said ACP protein ...

Compositions and Methods of Use for Recombinant Human Secretoglobins

Methods of synthetically producing, formulating and using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 are provided. Methods of using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 as therapeutic agents to affect long term patient outcomes, such as preventing severe respiratory exacerbations of underlying conditions that require medical intervention, including hospitalization are provided. Methods of producing recombinant human secretoglobins, analytical methods, pharmaceutical compositions, and methods of use to prevent the long term sequelae of acute and chronic respiratory conditions are provided. 16-. (canceled)7. A method of preventing hospitalization due to a severe respiratory exacerbation in a patient with acute lung injury comprising the steps of:(a) administering rhSCGB3A2 to a patient having an acute lung injury,wherein the patient is not re-hospitalized for at least ten months after administration of rhCC10.8. A method of preventing hospitalization due to a severe respiratory exacerbation in a patient with who experiences frequent respiratory exacerbations comprising the steps of:(a) administering rhSCGB3A2 to a patient having frequent respiratory exacerbations,wherein the patient is not re-hospitalized for at least two months after administration of rhSCGB3A2.9. A method of preventing hospitalization due to a severe respiratory exacerbation in a patient with a chronic respiratory condition comprising the steps of:(a) administering rhSCGB3A2 to a patient having a chronic respiratory condition,wherein the patient is not re-hospitalized for at least one month after administration of rhSCGB3A2.10. The method of wherein the severe respiratory exacerbation is caused by pulmonary fibrosis or bronchiectasis.11. The method of wherein the severe respiratory exacerbation is caused by pulmonary fibrosis or bronchiectasis.12. The method of wherein the severe respiratory exacerbation is caused by pulmonary fibrosis or bronchiectasis.1318.-. (canceled) This application ...

POLYPEPTIDES HAVING PROTEASE ACTIVITY

The present invention relates to isolated polypeptides having protease activity and isolated nucleic acid sequences encoding the proteases. The invention also relates to nucleic acid constructs, vectors, and host cells, including plant and animal cells, comprising the nucleic acid sequences, as well as methods for producing and using the proteases, in particular the use of the proteases in animal feed. 125-. (canceled)26. An isolated polypeptide having protease activity , selected from the group consisting of:(a) a polypeptide having at least 84% sequence identity to the polypeptide of SEQ ID NO: 5, SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 2 or SEQ ID NO: 4;(b) a polypeptide having at least 83% sequence identity to the polypeptide of SEQ ID NO: 19, or the mature polypeptide of SEQ ID NO: 16 or SEQ ID NO: 18;(c) a polypeptide having at least 85% sequence identity to the polypeptide of SEQ ID NO: 24, or the mature polypeptide of SEQ ID NO: 21 or SEQ ID NO: 23; (i) the mature polypeptide coding sequence of SEQ ID NO: 1,', '(ii) the mature polypeptide coding sequence of SEQ ID NO: 3,', '(iii) the mature polypeptide coding sequence of SEQ ID NO: 15,', '(iv) the mature polypeptide coding sequence of SEQ ID NO: 17,', '(v) the mature polypeptide coding sequence of SEQ ID NO: 20,', '(vi) the mature polypeptide coding sequence of SEQ ID NO: 22,', '(vii) the full-length complementary strand of (i), (ii), (iii), (iv), (v) or (vi);, '(d) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions, or very high stringency conditions with'}(e) a polypeptide encoded by a polynucleotide having at least 84% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or SEQ ID NO: 3;(f) a polypeptide encoded by a polynucleotide having at least 83% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 15 or SEQ ID NO: 17;(g) a polypeptide encoded by a polynucleotide having at least 85% sequence identity to ...

METHOD FOR SYNTHESIZING PROTEIN NANOPARTICLES USING WASTE CHICKEN FEATHERS

The embodiments herein disclose a method of synthesizing protein nanoparticles from waste chicken feathers by enzymatic hydrolysis followed by ultrasonic treatment. The steps for the synthesis of the protein nanoparticles include pretreatment of the chicken feathers. The next step is hydrolyzing the feather fibres enzymatically. Further the effect of the enzyme concentration, hydrolysis time and substrate concentration are analyzed. Also the protein nanoparticles are characterized. The effects of enzyme concentration, hydrolysis time, and substrate concentration on particle mean size are analyzed to optimize the best condition in order to attain the smallest particles by a Box-Behnken Design. It was found that minimum particle size can be obtained by using 5 g/l feather and 3.6% enzyme at hydrolysis time of 243 h. A validation assay confirmed the predictive response value under the optimal conditions. 1. A method of synthesizing protein nanoparticles from waste chicken feathers , the method comprising steps of:pretreating the plurality of waste chicken feathers;hydrolyzing the plurality of waste chicken feather fibers enzymatically;analyzing the effects of an enzyme concentration, a hydrolysis time and a substrate concentration; andsynthesizing the feather nanoparticles according to a particle shape, a particle size, crystallinity index and thermal stability characteristics.2. The method according to claim 1 , wherein the steps of pretreating the chicken feathers comprises:washing the plurality of waste chicken feathers in an aqueous solution for 30 minutes at 60° C., and wherein the aqueous solution comprises 1 g/L of a non-ionic detergent and 1 weight % of sodium carbonate at liquid to fiber ratio of 40 ml/g;rinsing the chicken feathers and drying;Soxhert extracting the chicken feathers by boiling the chicken feathers in petroleum ether for 12 hours to remove grease to obtain de-fattened feather fibers, and wherein the chicken feathers are boiled in petroleum ...

Compositions and Methods of Use for Recombinant Human Secretoglobins

Methods of synthetically producing, formulating and using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 are provided. Methods of using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 as therapeutic agents to affect long term patient outcomes, such as preventing severe respiratory exacerbations of underlying conditions that require medical intervention, including hospitalization are provided. Methods of producing recombinant human secretoglobins, analytical methods, pharmaceutical compositions, and methods of use to prevent the long term sequelae of acute and chronic respiratory conditions are provided. 1. A method of preventing hospitalization due to a severe respiratory exacerbation in a patient with acute lung injury comprising the steps of:administering recombinant human SCGB3A2 (rhSCGB3A2) protein, wherein the rhSCGB3A2 protein consists of the amino acid sequence set forth in SEQ ID NO: 3, to a patient having an acute lung injury,wherein the patient is not re-hospitalized for at least ten months after administration of rhSCGB3A2.2. A method of preventing hospitalization due to a severe respiratory exacerbation in a patient who experiences frequent respiratory exacerbations comprising the steps of:administering recombinant human SCGB3A2 (rhSCGB3A2) protein, wherein the rhSCGB3A2 protein consists of the amino acid sequence set forth in SEQ ID NO: 3, to a patient having frequent respiratory exacerbations,wherein the patient is not re-hospitalized for at least two months after administration of rhSCGB3A2.3. A method of preventing hospitalization due to a severe respiratory exacerbation in a patient with a chronic respiratory condition comprising the steps of:administering recombinant human SCGB3A2 (rhSCGB3A2) protein, wherein the rhSCGB3A2 protein consists of the amino acid sequence set forth in SEQ ID NO: 3, to a patient having a chronic respiratory condition,wherein the patient is not re-hospitalized for at least one month after administration of rhSCGB3A2.4. The ...

METHOD FOR PURIFYING TARGET PROTEIN

Disclosed is a method for purifying a target protein, comprising steps of: bringing a fusion protein containing an amino acid sequence of a peptide tag, an amino acid sequence of a cleavable site of a protease and an amino acid sequence of a target protein, into contact with the protease in a solution, to cleave the peptide tag from the fusion protein; and bringing the solution containing the peptide tag, the target protein and the protease into contact with an ion exchange resin to separate the target protein and the peptide tag, thereby acquiring a solution containing the target protein, wherein, in the fusion protein, the amino acid sequence of a cleavable site of the protease exists between the amino acid sequence of the peptide tag and the amino acid sequence of the target protein, and the peptide tag is polyanionic or polycationic. 1. A method for purifying a target protein , comprising steps ofbringing a fusion protein comprising an amino acid sequence of a peptide tag, an amino acid sequence of a cleavable site of a protease and an amino acid sequence of a target protein, into contact with the protease in a solution, to cleave the peptide tag from the fusion protein, andbringing the solution comprising the peptide tag, the target protein and the protease into contact with an ion exchange resin to separate the target protein and the peptide tag, thereby obtaining a solution comprising the target protein,whereinin the fusion protein, the amino acid sequence of a cleavable site is located between the amino acid sequence of the peptide tag and the amino acid sequence of the target protein, andthe peptide tag is polyanionic or polycationic.2. The method for purifying a target protein according to claim 1 , wherein the peptide tag is polyanionic claim 1 , and the ion exchange resin is an anion exchange resin.3. The method for purifying a target protein according to claim 1 , wherein the polyanionic peptide tag comprises 2 or more acidic amino acid residues.4. The ...

Portable Fluidic Platform For Rapid Cell-Free Production of Protein Biologics

A portable fluidic platform for rapid and flexible end-to-end production of recombinant protein biologics includes a bioreactor system hosting stable and robust cell-free translation systems that is fluidically integrated with modular protein separation functionalities (e.g., size exclusion, ion exchange or affinity chromatography systems) for purification of the cell-free expressed product and which are configurable for process-specific isolation of different proteins, as well as for formulation. The bioreactor utilizes lysates from engineered eukaryotic (e.g., yeast) or prokaryotic (e.g., bacterial) strains that contain factors for protein folding and posttranslational modifications. Combination of various purification modules on the same fluidic platform allows flexibility of re-routing for purification of different proteins depending on specific target requirements. Protein synthesis and purification modules are integrated into self-contained disposable fluidic cartridge that eliminates cross-contamination between runs. The platform allows for flexible production of protein biologics within 24 hours (from DNA to purified product). 1. A cell-free method for producing an amount of a protein biologic formulation at pharmaceutical dosage levels at a point of need , the method comprising:loading by a loading module a reaction solution including one or more reaction reagents to a reactor module;loading by a feeder module a feeding solution to the reactor module, the reactor module having a first chamber and a second chamber divided by a membrane;receiving at the first chamber the one or more reaction agents from the loading module;receiving at the second chamber the feeding solution from the feeder module;producing by the reactor module a target protein via a process selected from the group consisting of: (i) continuous exchange cell-free (CECF) and (ii) batch protein synthesis;separating by a chromatography module the target protein from one or more components of the ...

METHOD OF PRODUCING A RECOMBINANT PEPTIDE

Methods of producing a recombinant peptide, such as vessel dilator peptide (VSDL), are disclosed, including a particular method involving expressing a fusion polypeptide comprising concatemeric repeats of the peptide wherein the peptide is flanked by peptide cleavage sites, and cleaving the fusion polypeptide at the peptide cleavage sites with a cleaving agent(s) so as to release the peptide from the fusion polypeptide. Expression constructs and host cells for producing the fusion polypeptide and purification methods for the recombinant peptide are also disclosed. 2. The method of claim 1 , whereinsaid fusion polypeptide comprises concatemeric copies of said peptide.3. (canceled)4. The method of claim 2 , wherein said peptide has a native sequence.5. The method of claim 2 , wherein said peptide comprises a vessel dilator (VSDL) peptide or a variant or modified peptide thereof.6. The method of claim 2 , wherein said peptide is a VSDL peptide comprising the amino acid sequence according to SEQ ID NO: 2.7. The method of claim 2 , wherein the peptide cleavage sites are trypsin cleavage sites.8. The method of claim 2 , wherein the cleaving agent is trypsin or a trypsin-like enzyme.9. The method of claim 2 , wherein said peptide comprises a vessel dilator (VSDL) peptide or a variant or modified peptide thereof claim 2 ,wherein the peptide cleavage sites are trypsin cleavage sites and wherein the cleaving agent is trypsin or a trypsin-like enzyme.10. The method of claim 9 , wherein the peptide is a VSDL peptide comprising the amino acid sequence according to SEQ ID NO: 2.11. (canceled)12. The method of claim 2 , further comprising the steps of:adjusting the pH of the cleaved fusion polypeptide to below 5; andpurifying the peptide from the digested fusion polypeptide using at least one anion exchange technique.13. The method of claim 2 , further comprising spray drying or freeze drying the peptide.14. The method of claim 2 , wherein the fusion polypeptide comprises from two ...

METHOD FOR THE PREPARATION OF A PROTEIN PEPTIDE, A PROTEIN PEPTIDE AND USE THEREOF

The present invention discloses a method for the preparation of a protein peptide, peptide powder prepared thereby, and use of the peptide powder in hypouricemic food products or health care products. The peptide powder is capable of inhibiting the activity of xanthine oxidase and effectively reducing uric acid. The method in an example of the present invention comprises: mincing the tuna, heating with steam or water to obtain a pretreated tuna, enzymolysing the pretreated tuna, deactivating the enzyme, centrifuging, removing impurities to obtain a supernatant, concentrating, and drying to obtain the peptide powder of interest. 1. A method for the preparation of a protein peptide , characterized in that comprising:(1) mincing tuna, heating after adding water or heating with water steam to obtain pretreated tuna;(2) enzymolysing the pretreated tuna, deactivating the enzyme, centrifuging to obtain a supernatant;(3) concentrating the supernatant and drying to obtain the peptide powder of interest, which is the protein peptide.2. The method according to claim 1 , characterized in that the step of heating after adding water or heating with steam comprises heating at 80 to 100° C. for 5 to 30 min with steam or water.3. The method according to claim 1 , characterized in that the steps of enzymolysis and deactivating the enzyme comprise: mixing the pretreated tuna with protease at a temperature of 50 to 60° C. for 3.0 to 9.0 h; deactivating the enzyme at a temperature of 90 to 100° C. for 10 to 30 min after completion of the reaction.4. The method according to claim 1 , characterized in that the protease used in enzymolysis is one or more of acid protease claim 1 , papain claim 1 , pepsin claim 1 , trypsin claim 1 , proteolytic enzyme claim 1 , neutrase claim 1 , flavourzyme and Alcalase claim 1 , and the protease is used in an amount of 0.2% to 3.2% by weight of the pretreated tuna.5. The method according to claim 1 , characterized in that further comprises removing ...

ISOLATED POLYPEPTIDES, KITS COMPRISING THE SAME AND USES THEREOF

Disclosed herein are isolated polypeptides, kits comprising the same, and uses thereof. The polypeptides are respectively isolated from and are capable of catalyzing the cleavage of a peptide linkage located between the C-terminus of the distal ubiquitin (UB) and any of the first methionine (M1), the 48(K48) or the 63lysine (K63) of the proximal UB in a UB chain, which is conjugated to a substrate protein. 1. An isolated polypeptide capable of cleaving a peptide linkage located between the C-terminus of the distal ubiquitin (UB) and any of the first methionine (M1) , the 48(K48) or the 63lysine (K63) of the proximal UB in a UB chain conjugated with a substrate protein , comprising:an amino acid sequence at least 85% identical to any of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9.2Arabidopsis thalianaA. thaliana. The isolated polypeptide of claim 1 , wherein the polypeptide is isolated from ().3. The isolated polypeptide of claim 2 , wherein the isolated polypeptide has the amino acid sequence at least 85% identical to SEQ ID NO: 1 claim 2 , and mutations at amino acid residues 92 claim 2 , 89 and 288 of the isolated polypeptide abolish its activity to cleave the peptide linkage located between the C-terminus of the distal UB and the M1 claim 2 , K48 claim 2 , or K63 of the proximal UB in the UB chain.4. The isolated polypeptide of claim 2 , wherein the isolated polypeptide has the amino acid sequence at least 85% identical to SEQ ID NO: 4 claim 2 , and a mutation at amino acid residue 136 of the isolated polypeptide abolish its activity to cleave the peptide linkage located between the C-terminus of the distal UB and the K48 or K63 of proximal UB in the UB chain.5. The isolated polypeptide of claim 4 , wherein the isolated polypeptide has the amino acid sequence of SEQ ID NO: 5.6. The isolated polypeptide of claim 2 , wherein the isolated polypeptide has the amino acid sequence at ...

METHODS FOR PRODUCING DIKETOPIPERAZINES AND COMPOSITIONS CONTAINING DIKETOPIPERAZINES

Methods of making increased amounts of diketopiperazines (DKP) such as DA-DKP in pharmaceutical compositions of proteins and peptides are disclosed. The disclosure further provides methods of making a DKP, including (1) contacting albumin with an enzyme (such as a dipeptidyl peptidase N (DPP-IV)) that cleaves a pair of N-terminal amino acids from the albumin, and (2 heating the albumin under conditions effective to cause the formation of the DKP. Further, treatment of DKP- and albumin-containing streams to produce improved, higher value, DKP compositions and purified albumin compositions for therapeutic uses is also disclosed. In addition to a first therapeutic DKP composition comprising a low albumin content, a second valuable therapeutic composition is also produced characterized by a high albumin concentration. 1. A method for treating a feed stream comprising albumin and aspartic acid-alanine diketopiperazine (DA-DKP) to produce compositions , the method comprising:processing the feed stream to produce a first albumin-lean stream and a first albumin-rich stream, wherein the first albumin-lean stream comprises a first portion of the DA-DKP present in the feed stream, and the first albumin-rich stream comprises a second portion of the DA-DKP present in the feed stream;reacting the first albumin-rich stream in order to produce DA-DKP resulting in a reaction stream comprising albumin and DA-DKP; andprocessing the reaction stream to produce a second albumin-lean stream and a second albumin-rich stream, wherein the second albumin-lean stream comprises a portion of the DA-DKP present in the reaction stream, and the second albumin-rich stream comprises a second portion of the DA-DKP present in the reaction stream.2. The method of claim 1 , wherein at least one of the first and second albumin-lean streams possess therapeutic value.3. The method of claim 1 , wherein at least one of the first and second albumin-rich streams possess therapeutic value.4. The method of claim ...

LIPASE VARIANTS AND POLYNUCLEOTIDES ENCODING SAME

The present invention relates to a lipase variant of a parent lipase, which variant has lipase activity, at least 75% but less than 100% sequence identity to SEQ ID NO: 3 and comprises a substitution at one or more positions corresponding to positions 1; 2; 3; 4; 5; 6; 7; 9; 10; 11; 12; 16; 19; 30; 31; 34; 36; 37; 39; 40; 42; 44; 51; 52; 53; 54; 56; 58; 59; 70; 71; 72; 73; 83; 84; 86; 88; 90; 92; 93; 95; 96; 100; 101; 102; 104; 106; 109; 110; 112; 117; 119; 124; 125; 127; 128; 131; 132; 133; 134; 135; 137; 158; 159; 160; 161; 162; 163; 165; 166; 167; 168; 170; 181; 182; 183; 189; 190; 192; 194; 196; 202; 210; 211; 212; 220; 225; 227; 228; 229; 230; 231; 233; 237; 238; 239; 240; 242; 246; 247; 248; 252; 259; 262; 264; 269 of SEQ ID NO: 3. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants. 1. A lipase variant which has lipase activity , at least 75% but less than 100% sequence identity to SEQ ID NO: 3 , and comprises an amino acid substitution at one or more positions corresponding to positions 1; 2; 3; 4; 5; 6; 7; 9; 10; 11; 12; 16; 19; 30; 31; 34; 36; 37; 39; 42; 44; 51; 52; 53; 54; 56; 58; 59; 70; 71; 72; 73; 83; 88; 92; 93; 95; 96; 100; 101; 102; 104; 106; 109; 110; 112; 117; 119; 124; 125; 127; 128; 131; 132; 133; 134; 135; 137; 158; 159; 160; 161; 162; 163; 165; 166; 167; 168; 170; 181; 182; 183; 189; 190; 192; 194; 196; 202; 210; 211; 212; 220; 225; 227; 228; 229; 230; 231; 233; 237; 238; 239; 240; 242; 246; 247; 248; 252; 259; 262; 264; 269 of SEQ ID NO: 3.2. The lipase variant of claim 1 , which comprises an amino acid substitution at one or more positions corresponding to positions 1(C claim 1 ,F claim 1 ,G claim 1 ,H claim 1 ,I claim 1 ,L claim 1 ,M claim 1 ,P claim 1 ,Q claim 1 ,R claim 1 ,V claim 1 ,W claim 1 ,Y); 2(A claim 1 ,C claim 1 ,D claim 1 ,M claim 1 ,N claim 1 ,P claim 1 ,Q claim 1 ,R claim 1 ,S claim 1 ,T ...

OYSTER PEPTIDE CAPABLE OF ENHANCING SEXUAL FUNCTION, AND PREPARATION METHOD AND APPLICATION THEREOF

An oyster peptide capable of enhancing a sexual function, and a preparation method and application thereof are provided. In the method, oyster meat is pre-treated by using calcium salt before enzymatic hydrolysis, so as to activate and release endogenous enzymes of oysters, such that enzymatic preparations consumed in subsequent enzymatic hydrolysis can be reduced. 1. A method for preparing a sexual function-improving oyster peptide , comprising:1) grinding oyster flesh, calcium salt and water to obtain oyster flesh pulp;2) enzymatic hydrolyzing oyster flesh pulp, centrifuging and collecting the supernatant to obtain an oyster enzyme-hydrolyzed raw solution;3) decoloring the oyster enzyme-hydrolyzed raw solution and removing impurities to obtain a fine oyster peptide solution; and4) concentrating and spay drying the fine oyster peptide solution to obtain the oyster peptide.2. The method according to claim 1 , wherein the calcium salt is food grade calcium chloride claim 1 , calcium lactate claim 1 , calcium carbonate claim 1 , calcium hydrogen phosphate or calcium citrate.3. The method according to claim 1 , wherein the amount of the calcium salt in step 1) is 0.1%˜0.3% of the oyster flesh mass.4. The method according to claim 1 , wherein the step 2) comprises:enzymatic hydrolyzing the oyster flesh pulp by neutral proteases or alkaline proteases;adding flavourzyme for further hydrolyzation;inactivating enzymes after enzymatic hydrolyzation; andcentrifuging and collecting the supernatant to obtain an oyster enzyme-hydrolyzed raw solution.5. The method according to claim 4 , wherein the step 2) comprises:stirring the oyster flesh pulp at 35° C.˜45° C. for 1 h˜2 h;adding neutral protease or alkaline protease;adjusting the pH and heat to 50° C.˜60° C. to continue hydrolysis for 5 h˜8 h;adjusting the pH to 5.0 to 5.5 and adding flavourzyme to continue hydrolysis at 50° C.˜60° C.;inactivating the enzymes 2 h—3 h after the hydrolysis; andcentrifuging and collecting the ...

Chemo-Enzymatic Site-Specific Modification of Peptides and Proteins to Form Cleavable Conjugates

A method is provided for reversibly modifying a protein or peptide on its glutamine residue(s) by performing a reaction, such as a transglutaminase-catalyzed reaction, between the protein or peptide and an amine-containing reagent, whereby the reagent is linked through its amine function to a side chain of the glutamine residue. Subjecting the modified protein to an appropriate stimulus regenerates the protein or peptide in its original form. 1. A method for modifying a protein or peptide having one or more glutamine residues , the method comprising:performing a transglutaminase-catalyzed reaction between the protein or peptide and an amine-containing reagent, thereby producing a first derivative of the protein or peptide, wherein the reagent becomes covalently linked through its amine function to a side chain of at least one of said one or more glutamine residues; wherein the original protein or peptide, or a second derivative thereof, can be obtained by a treatment of the first derivative of the protein or peptide.2. The method of claim 1 , further comprising carrying out said treatment of the first derivative.3. The method of claim 1 , wherein the treatment comprises a photolysis reaction.5. The method of claim 3 , wherein the amine-containing reagent comprises: (1) a hydroxylamine group claim 3 , and the reagent is linked through the hydroxylamine group to the side chain of the at least one glutamine residue; or (2) a hydrazine group claim 3 , and the reagent is linked through the hydrazine group to the side chain of the at least one glutamine residue.8. The method of claim 1 , wherein the treatment comprises a reaction catalyzed by a metal in elemental form claim 1 , a metal in ionic form claim 1 , or a metal that is part of a complex.10. The method of claim 8 , wherein the amine-containing reagent is a reagent comprising a hydroxylamine group claim 8 , wherein the reagent is linked through the hydroxylamine group to a side chain of the at least one glutamine ...

LOW CROSS-LINKING GELATINE

The present invention relates to a process for the preparation of a gelatine blend comprising the steps of a) providing a gelatine extract or a series of gelatine extracts, differing in bloom and viscosity; b) subjecting a portion of one or more of the gelatine extracts of a) to an enzymatic treatment to obtain one or more enzymatically treated extracts having a viscosity that is lower than that of the respective extract before said enzymatic treatment, while maintaining the bloom substantially unaffected, the enzymatically treated extracts comprising less than 1.5 w/w % of gelatine-derived peptides having a molecular weight of less than 2000 Da; c) selecting one or more gelatine extracts of step a); d) combine the selected one or more gelatine extracts of step c) and blend with one or more enzymatically treated extracts of step b) to provide a gelatine composition having a lower viscosity as compared to that of the said combined selected extracts of step c) without being blended with said selected one or more enzymatically treated gelatine extracts of step b), while maintaining the bloom substantially unaffected. The present invention also relates to a gelatine composition, and a gelatine product comprising said gelatine composition. 1. Process for the preparation of a gelatine blend comprising the steps of:a) providing a gelatine extract or a series of gelatine extracts, differing in bloom and viscosity,b) subjecting a portion of one or more of the gelatine extracts of step a) to an enzymatic treatment to obtain one or more enzymatically treated extracts having a viscosity that is lower than that of the respective extract before said enzymatic treatment, while maintaining the bloom substantially unaffected, the enzymatically treated extracts comprising less than 1.5 w/w % of gelatine-derived peptides having a molecular weight of less than 2000 Da,c) selecting one or more gelatine extracts of step a),d) combine the selected one or more gelatine extracts of step c) ...

SELF-LABELING MINIPROTEINS AND CONJUGATES COMPRISING THEM

Disclosed are reactive miniproteins, and peptides comprising them. The miniproteins and peptides are amino acid sequences not found in nature that are able to undergo SAr chemistry and other nucleophilic based reactions. Also disclosed are conjugates comprising at least one of the reactive miniproteins or peptides, and methods of forming these conjugates. 1. A peptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-158 , or a peptide consisting of an amino acid sequence having at least 95% homology with a peptide selected from the group consisting of SEQ ID NOs: 1-158.2. The peptide of claim 1 , wherein the peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-34.3. The peptide of claim 1 , wherein the peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 35-59.4. The peptide of claim 1 , wherein the peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 60-158.5. The peptide of claim 1 , wherein the peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 12 claim 1 , 13 claim 1 , 61-64 claim 1 , 66 claim 1 , 67 claim 1 , 69-71 claim 1 , 81-113 claim 1 , and 115-158.6. The peptide of claim 1 , wherein the peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 12 claim 1 , 13 claim 1 , 61-64 claim 1 , 66 claim 1 , 67 claim 1 , 69-71 claim 1 , 81 claim 1 , 82 claim 1 , 90 claim 1 , 97 claim 1 , 98 claim 1 , 104 claim 1 , 106-113 claim 1 , 115-123 claim 1 , 133 claim 1 , and 144-158.7. The peptide of claim 1 , wherein the peptide consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 124-132 and 136-143.8. The peptide of claim 1 , wherein the peptide consists of an amino acid sequence having 1 claim 1 , 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , or 6 point mutations compared to a peptide of SEQ ID NO: 13. ...

一种抗左旋氧氟沙星抗体的F(ab)2片段及其制备方法和应用

本发明公开了一种抗左旋氧氟沙星抗体的F(ab) 2 片段，所述F(ab) 2 片段具有通过二硫键连接在一起的两个Fab部分，每个Fab部分包含轻链和重链Fd部分，所述轻链的氨基酸序列如SEQ ID NO:3所示，所述重链Fd部分如SEQ ID NO:4所示。本发明通过胃蛋白酶酶解，采用两步纯化法纯化酶解产物，首次获得抗左旋氧氟沙星抗体F(ab) 2 。本发明方法流程简洁，操作简单，高产出，所述抗左旋氧氟沙星抗体的F(ab) 2 片段对左旋氧氟沙星的半抑制浓度为0.33ng/mL，检测限为0.04ng/mL，检测范围为0.05～2.15ng/mL，具有较大的应用前景。

一种羊乳乳清高f值寡肽的制备方法

本发明涉及高F值寡肽制备技术领域，尤其涉及一种羊乳乳清高F值寡肽的制备方法。所述方法包括步骤：(1)将羊乳依次进行脱脂、脱酪蛋白、脱小分子物质后冻干，制得乳清蛋白粉；(2)对所述乳清蛋白粉依次进行酶解、脱芳、离心过滤，得到羊乳乳清高F值寡肽液，即得。本发明使用两种酶分步水解，再使用活性炭将芳香族氨基酸去除，本发明得到了一种无异味，透明液体状的羊乳乳清高F值寡肽，其F值高达26.32，是一种高支链氨基酸低芳香族氨基酸的寡肽体系。

一种羊胎盘活性肽的制备方法

本发明涉及多肽技术领域，具体公开一种羊胎盘活性肽的制备方法。所述制备方法，包括如下步骤：选取新鲜羊胎盘，经粉碎处理得到胎盘浆液；加入脂肪酶、无花果蛋白酶及复合风味蛋白酶，酶解得到酶解液；酶解液经过滤、离心脱脂、纳滤、脱盐和浓缩、离子纯化、冷冻干燥，得到羊胎盘活性肽。本发明提供的羊胎盘活性肽的制备方法，提高水解度，使高分子多肽水解为低分子的活性肽，得到高浓度、高活性且分子量均一性好的低分子量的生物活性肽。

Application of protein hydrolysates for stabilisation of detergent compositions of metalloprotease

FIELD: biotechnologies. SUBSTANCE: composition of neutral metalloprotease stabilised with an inhibitor is proposed to produce a liquid detergent solution. The composition contains from approximately 0.001% to approximately 10% wt of neutral metalloprotease and a competitive inhibitor. Besides, the competitive inhibitor represents a protein hydrolysate and is connected with at least approximately 90% of molecules of the specified neutral metalloprotease. Also the method is proposed to produce a composition of neutral metalloprotease stabilised with an inhibitor. The mixture is incubated, which contains at least one neutral metalloprotease and a protein substrate, in the water buffer at pH in the range from approximately 6.5 to approximately 11 and at the temperature from approximately 22°C to approximately 37°C. In process of incubation the substrate protein is split when exposed to metalloprotease, which results in formation of the hydrolysis product. The hydrolysis product is extracted with molecular weight of less than approximately 5000 Da and combined with neutral metalloprotease. EFFECT: higher stability of detergent compositions during storage without reduction of desired activity of neutral metalloprotease in process of application. 12 cl, 3 dwg, 5 tbl, 3 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) 2 484 138 (13) C2 (51) МПК C12N C11D C11D 9/96 (2006.01) 3/38 (2006.01) 3/386 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (21)(22) Заявка: 2009144091/10, 23.04.2008 (24) Дата начала отсчета срока действия патента: 23.04.2008 (72) Автор(ы): ЛИ Санг-Киу (US), ВИНЕЦКИ Дебора С. (US) (43) Дата публикации заявки: 10.06.2011 Бюл. № 16 2 4 8 4 1 3 8 (45) Опубликовано: 10.06.2013 Бюл. № 16 2 4 8 4 1 3 8 R U (85) Дата начала рассмотрения заявки PCT на национальной фазе: 30.11.2009 C 2 C 2 (56) Список документов, цитированных в отчете о поиске: WO 2007044993 А2, 19.04.2007. KAMMOUN R. ET AL. Protein size distribution and ...

一种用水蛭制取降血脂肽的方法

本发明公开了一种用水蛭制取降血脂肽的方法，包括将天然水蛭净化、磨浆、酶解、灭酶、离心分离、超滤和冷冻干燥，最终得到的水蛭制取广州旭远照明科技有限公司降血脂肽。本发明的有益效果：降血脂肽是由65～66氨基酸组成的低分子多肽，其分子量仅为7000道尔顿，极易溶于水和生理盐水，在常温下非常稳定，单纯的温度升高(100℃水浴)，或pH改变(1～13)均不影响其活性。

A kind of preparation method of spleen aminopeptide

本发明涉及动物脾脏的深加工技术领域，尤其涉及一种脾氨肽的制备方法。该方法采用新鲜牲畜脾脏为主要原料，经过前处理、酶解、固液分离、脱脂纯化、纳滤、浓缩纯化，得到脾氨肽的浓缩液。该浓缩液可经冻干后即得到冻干产品。本发明所提供的制备方法生产周期短、成本低、对环境无污染，而且所得产品安全性高，纯度高、活性高、分子量可控，可广泛应用于食品、特医食品、保健品、药品等领域。

A kind of efficient preparation process of the anti-oxidant calcium ion chelating peptide in Optimization of Low Value Fish visceral protein source

本发明公开了一种低值鱼内脏蛋白源抗氧化钙离子螯合肽的高效制备工艺，涉及微波预处理；超声波辅助酶解；超滤处理；CaCl 2 螯合；以及凝胶柱处理；本发明通过Ca 2+ 与抗氧化肽相结合，提高了抗氧化肽的结构稳定性，形成抗氧化钙离子螯合肽，具有较强的抗氧化活性以及在加工和贮藏过程中的抗氧化稳定性，并以金线鱼内脏为原料，来源广，成本低，解决了鱼糜加工过程中产生副产物的问题，对进一步提高金线鱼的经济价值值具有的理论意义；此外，本发明通过微波处理金线鱼内脏蛋白，降低蛋白中的有序结构并提高其的溶解性，在此基础上，将超声波处理和蛋白酶酶解结合，提高水解度和得率；同时通过多次分离纯化、获得可靠的金线鱼内脏抗氧化钙离子螯合肽。

Method for extracting natural antimicrobial peptides from leukocyte-erythrocyte-platelet blood mass

FIELD: biotechnology. SUBSTANCE: invention relates to the field of biotechnology. Disclosed is a method of extracting natural antimicrobial peptides (AMP) from leukocyte-erythrocyte-platelet blood mass. Used is a leukocyte-erythrocyte-platelet blood mass pre-exposed to trypsin haemolysis. Separation of AMP is carried out by liquid chromatography on a separating column with three-fold use of Sephadex G-25. EFFECT: invention enables to optimize the technology of extracting the most complete fraction of natural low-molecular weight peptides containing AMP, to use Sephadex G-25 several times after washing, regeneration and drying, to increase their biological value and to reduce the cost of production thereof. 1 cl, 2 tbl, 3 dwg РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 729 016 C1 (51) МПК C07K 1/12 (2006.01) C12P 21/06 (2006.01) G01N 30/02 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C07K 1/12 (2020.02); C12P 21/06 (2020.02); G01N 30/02 (2020.02) (21)(22) Заявка: 2019134760, 29.10.2019 (24) Дата начала отсчета срока действия патента: Дата регистрации: 04.08.2020 (45) Опубликовано: 04.08.2020 Бюл. № 22 Адрес для переписки: 355017, г. Ставрополь, ул. Мира, 310, Медуниверситет, Научно-аналитический отдел, Молдовановой Л.С. 2 7 2 9 0 1 6 C 1 (56) Список документов, цитированных в отчете о поиске: RU 2416243 C2, 20.04.2011. RU 2510398 C2, 27.03.2014. JABEEN U. et al. Isolation and characterization of potential food preservative peptide from Momordica charantia L. Arabian Journal of Chemistry. May 2017; 10(Supplement 2): S3982-S3989. DAHOT M.U. Antimicrobial activity of small protein of Moringa oleifera leaves. Journal of Islamic Academy of (см. прод.) (54) СПОСОБ ВЫДЕЛЕНИЯ ПРИРОДНЫХ АНТИМИКРОБНЫХ ПЕПТИДОВ ИЗ ЛЕЙКОЦИТАРНО-ЭРИТРОЦИТАРНО-ТРОМБОЦИТАРНОЙ МАССЫ КРОВИ (57) Реферат: Изобретение относится к области колонке с трехкратным использованием биотехнологии. Предложен способ выделения Сефадекса G-25. Изобретение ...

A kind of preparation method of silver carp fish protein peptide

本发明公开了一种白鲢鱼蛋白肽的制备方法，包括准备、第一次酶解、第二次酶解、灭活、过滤去粗渣、高速离心脱脂、脱色去味、干燥八个步骤。本发明工艺流程相对简单且原材料易得，第一步酶解后无需升温灭活及再降温，简化了工艺，适合大工业化生产；且制得的白鲢鱼蛋白肽是纯天然营养物质，为低分子量多肽，人体极易吸收，含有人体所需的8种必须氨基酸，不但可以维持人体健康，还可以促进生长发育，是高档营养食品、保健食品。

Preparation method of silver carp protein peptide

本发明公开了一种白鲢鱼蛋白肽的制备方法，包括准备、第一次酶解、第二次酶解、灭活、过滤去粗渣、高速离心脱脂、脱色去味、干燥八个步骤。本发明工艺流程相对简单且原材料易得，第一步酶解后无需升温灭活及再降温，简化了工艺，适合大工业化生产；且制得的白鲢鱼蛋白肽是纯天然营养物质，为低分子量多肽，人体极易吸收，含有人体所需的8种必须氨基酸，不但可以维持人体健康，还可以促进生长发育，是高档营养食品、保健食品。

A kind of method that biomimetic method prepares functional protein polypeptide

本发明涉及一种仿生法制备功能性蛋白多肽的方法，是以天然植物蛋白为原料，利用非动物源性蛋白酶，模拟动物消化过程，生产酶解蛋白多肽。包括：将植物蛋白原料预粉碎，加水调制，采用非动物源性复合蛋白酶A、木瓜蛋白酶和菠萝蛋白酶进行一次酶解，再加入复合蛋白酶B、赖氨酸氨肽酶和羧肽酶进行二次酶解，制得二次酶解蛋白浆液，然后均质、喷雾干燥、灭菌，即得。本发明工艺流程简单，成本低，所制备的产品活性小分子肽(相对分子量介于186～1000Da之间)和游离氨基酸(<186Da)含量高，抗营养因子全部降解，体外消化吸收率≥95％以上，能有效替代优质动物源性蛋白，显著促进幼龄动物的生长，具有良好的社会和经济效益。

Segregated folding determinants for small disulfide-rich peptides

The preparation of small peptides with multiple disulfide bonds is accomplished by forming a prepropeptide with an N-terminal excised region separated from the cysteine-rich peptide by one or more cleavable amino acid residues. The excised region preferably consists of an N-terminal end providing a hydrophobic signal sequence domain having up to approximately 25 amino acids, and an intermediate central propeptide domain having a variable length of between about 5-50 amino acids. The N-terminal excised region serves as a folding template to direct the formation of specific disulfide bonds in the cysteine-rich peptide. The cysteine-rich peptide is cleaved by enzymes releasing the biologically active peptide.

Lipase variants and polynucleotides encoding same

The present invention relates to a lipase variant of a parent lipase, which variant has lipase activity, at least 75% but less than 100% sequence identity to SEQ ID NO: 3 and comprises a substitution at one or more positions corresponding to positions 1; 2; 3; 4; 5; 6; 7; 9; 10; 11; 12; 16; 19; 30; 31; 34; 36; 37; 39; 40; 42; 44; 51; 52; 53; 54; 56; 58; 59; 70; 71; 72; 73; 83; 84; 86; 88; 90; 92; 93; 95; 96; 100; 101; 102; 104; 106; 109; 110; 112; 117; 119; 124; 125; 127; 128; 131; 132; 133; 134; 135; 137; 158; 159; 160; 161; 162; 163; 165; 166; 167; 168; 170; 181; 182; 183; 189; 190; 192; 194; 196; 202; 210; 211; 212; 220; 225; 227; 228; 229; 230; 231; 233; 237; 238; 239; 240; 242; 246; 247; 248; 252; 259; 262; 264; 269 of SEQ ID NO: 3. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Lipase variants and polynucleotides encoding same

The present invention relates to a lipase variant of a parent lipase, which variant has lipase activity, at least 75% but less than 100% sequence identity to SEQ ID NO: 3 and comprises a substitution at one or more positions corresponding to positions 125; 1; 2; 3; 4; 5; 6; 7; 9; 10; 11; 12; 16; 19; 30; 31; 34; 36; 37; 39; 40; 42; 44; 51; 52; 53; 54; 56; 58; 59; 70; 71; 72; 73; 83; 84; 86; 88; 90; 92; 93; 95; 96; 100; 101; 102; 104; 106; 109; 110; 112; 117; 119; 124; 127; 128; 131; 132; 133; 134; 135; 137; 158; 159; 160; 161; 162; 163; 165; 166; 167; 168; 170; 181; 182; 183; 189; 190; 192; 194; 196; 202; 210; 211; 212; 220; 225; 227; 228; 229; 230; 231; 233; 237; 238; 239; 240; 242; 246; 247; 248; 252; 259; 262; 264; 269 of SEQ ID NO: 3. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Separation and extraction method of corn peptide for anti-alcohol beverage

一种用于解酒饮料的玉米肽的分离提取方法，经过预处理、酶水解、精制、浓缩、干燥六个步骤提取玉米肽，本发明通过加入有机溶剂无水乙醇将玉米蛋白粉中的蛋白质提取出来，搅拌，离心对蛋白粉进行提纯，同时无水乙醇可更好的减少玉米蛋白粉中的色素、脂肪的含量，使得下述步骤中的酶水解效果更佳，避免碱性水解酶的浪费；玉米蛋白粉结构紧密，不易被蛋白酶活性部位接触而讲解，因为在酶水解之前采用高温预处理破坏其内部空间结构，从而使得酶更好的作用于其作用位点进行讲解，减少水解时间，提高水解效率；该种提取方法最终形成的玉米肽粉，可以直接与其余材料混合形成口感适宜的解酒饮料，方便实用，同时也使得口感更加，易于被人体吸收。

Preparation method of aquatic protein and plant protein compound active peptide

本发明公开了一种水产蛋白-植物蛋白复合型活性肽的制备方法，包括以下主要步骤：将新鲜的水产蛋白绞碎后备用，将植物蛋白粉碎后过60目筛备用；绞碎后的水产蛋白和粉碎的植物饲料蛋白混合，并添加营养物和水混匀作为发酵培养基；往发酵培养基上接入能产耐高温蛋白酶的枯草芽孢杆菌（<i>Bacillus？subtilis</i>）HL-2；接入菌种的培养基于50℃～55℃进行固态好氧发酵38h～48h；然后固态发酵产物加水于55℃酶解2h；液态酶解产物先后经过滤、超滤和纳滤后得到浓缩液，浓缩液经冷冻干燥后得到活性肽成品。本发明技术作用条件温和，更多地保留了制备的功能肽和原料本身所含其它功能性成分的生物学活性，并且简化了制备工序和降低了活性肽的生产成本，有着良好的应用前景。

Fab fragment of anti-dextro-ofloxacin antibody, preparation and application thereof

本发明提供一种抗右旋氧氟沙星抗体的Fab片段及其制备与应用，Fab片段包含重链，所述重链含有如SEQ ID NO:2所示氨基酸序列或与SEQ ID NO:2具有至少80％同源性的氨基酸序列，或者所述重链的氨基酸序列如SEQ ID NO:2所示。本发明的Fab片段纯化流程简洁，操作简单，高产出，制得的抗右旋氧氟沙星Fab片段用于Elisa检测时，灵敏度高。

Use of protein hydrolysates to stabilize metalloprotease detergent formulations

본 발명은 증가된 보관 안정성을 보여주는 메탈로프로테아제 효소 및 단백질 가수분해물 억제제를 포함하는 조성물 및 제제를 제공한다. 한 구현예에서, 본 발명은 세제 제제에 단백질 가수분해물을 포함하여 안정화된, 하나 이상의 메탈로프로테아제 (예를 들어, 바실러스 sp. 중성 메탈로프로테아제) 를 포함하는 액체 세제 제제를 제공한다. 본 발명은 또한, 메탈로프로테아제 효소를 사용해 단백질 기질을 분해함으로써 세제 제제를 안정화시키기 위한 단백질 가수분해물의 제조 방법을 제공한다. 메탈로프로테아제 The present invention provides compositions and formulations comprising a metalloprotease enzyme and a protein hydrolyzate inhibitor exhibiting increased storage stability. In one embodiment, the present invention provides a liquid detergent formulation comprising one or more metalloproteases (e.g., Bacillus sp. Neutral metalloprotease) stabilized with a protein hydrolyzate in a detergent formulation. The present invention also provides a method for producing a protein hydrolyzate for stabilizing a detergent formulation by decomposing a protein substrate using a metalloprotease enzyme. Metalloprotease

Method for enzymatic production of GLP-2(1-33) and GLP-2(1-34) peptides

The invention provides methods for making peptides from a polypeptide containing at least one copy of the peptide using clostripain to excise the peptide from the polypeptide. The methods enable the use of a single, highly efficient enzymatic cleavage to produce any desired peptide sequence.

Method for enzymatic production of GLP-2(1-33) and GLP-2(1-34) peptides

The invention provides methods for making peptides from a polypeptide containing at least one copy of the peptide using clostripain to excise the peptide from the polypeptide. The methods enable the use of a single, highly efficient enzymatic cleavage to produce any desired peptide sequence.

Lipase variants and polynucleotides encoding same

Preparation method of black fungus active peptide

一种黑木耳活性肽的制备方法，它属于黑木耳小分子肽制备方法领域。本发明将黑木耳进行前处理后，进行三步酶解法，称量黑曲霉的纤维素酶和真菌淀粉酶进行酶的活化，活化后的酶加入发酵罐中，进行酶解反应后，灭酶倒入干净的发酵罐中，再称量食用菌水解酶、消化酶、大豆多肽水解酶、植物蛋白水解酶进行酶的活化，活化后的酶加入的发酵罐中，进行酶解反应后，倒入干净的发酵罐中，再称量木瓜蛋白酶、菠萝蛋白酶、碱性蛋白酶、胰蛋白酶进行酶的活化，活化后的酶加入发酵罐中，进行酶解反应后，得到黑木耳酶解液，进行减压蒸馏，得到的浓缩液进行喷雾干燥，以一定的料液比溶于磁化水中，得到黑木耳活性肽液。本发明为澄清淡黄色液体。

A kind of preparation method of leech active oligopeptides

本申请公开了一种水蛭活性寡肽的制备方法，包括：采用碱性蛋白酶和胰蛋白酶对水蛭进行酶解，得到所述水蛭活性寡肽；其中，所述水蛭活性寡肽的分子量低于500Da。本申请中所述方法解决了现有技术中水蛭多肽提取物制备方法难以保证多肽分子量较小及分子量分布集中的技术问题。

Technique for preparing squid skin collagen peptide through enzymic method

本发明涉及一种酶法制备鱿鱼皮胶原蛋白肽的工艺方法，原料鱿鱼皮预处理后，先经碱性蛋白酶水解后，再经复合蛋白酶继续水解得到胶原蛋白水解液，其中，复合蛋白酶由木瓜蛋白酶与风味蛋白酶组成。本发明工艺条件下制备得到的胶原蛋白肽产品，不仅水解度、DPPH清除率、多肽转化率等均较为理想，而且97.63％胶原蛋白肽分子量小于2000Da，具有较好的品质和较高的利用价值；本发明对鱿鱼皮胶原蛋白肽后期进一步的分离纯化及应用具有较高的理论指导意义。