СПОСОБ ВЫДЕЛЕНИЯ И ОЧИСТКИ 1,4-ДИАМИНОБУТАНА ИЗ ФЕРМЕНТАЦИОННОГО РАСТВОРА

Настоящее изобретение относится к биотехнологии. Предложены способы выделения и очистки 1,4-диаминобутана из ферментационного раствора. Предложенный способ выделения и очистки 1,4-диаминобутана из ферментационного раствора, содержащего 1,4-диаминобутан, полученного посредством ферментации, включает стадии отделения клеточной массы, добавления щелочного вещества и удаления образовавшихся солей, концентрирования, удаления примесей и извлечения полученного 1,4-диаминобутана. Также предложен способ выделения и очистки 1,4-диаминобутана из ферментационного раствора, содержащего 1,4-диаминобутан, полученного посредством ферментации, включающий отделение клеточной массы, добавление щелочного вещества и удаление образовавшихся солей, низкотемпературное концентрирование, кристаллизацию, фильтрацию, высокотемпературное концентрирование и дистилляцию. Предложенные способы выделения и очистки позволяют получить 1,4-диаминобутан методами биотехнологии из ферментационного раствора с высоким выходом и ...

СПОСОБ ПРОДУЦИРОВАНИЯ ОРГАНИЧЕСКИ СВЯЗАННОГО ВИТАМИНА В

Изобретение относится к биотехнологии. Настоящее изобретение относится к способу конверсии витамина группы B в растениях, предпочтительно выбираемого из витамина B1, B2, B3, B5, B6, B7, B9, B12 или их смесей, из синтетической формы в органически связанную, в котором всхожие семена растений сначала замачивают в растворе соответствующего витамина или смеси витаминов и в процессе прорастания опрыскивают растворами, содержащими соответствующий витамин. Изобретение позволяет получить витамины группы В с улучшенной биодоступностью. 2 н. и 17 з.п. ф-лы, 8 табл., 4 пр.

ПОЛИАМИДНАЯ СМОЛА И ШАРНИРНЫЕ ФОРМОВАННЫЕ ИЗДЕЛИЯ

Изобретение относится к вариантам полиамидной смолы в виде гранул, к композиции полиамидной смолы, к формованному изделию, к вариантам шарнирного формованного изделия, к бандажной ленте, к элементарной нити. По первому варианту полиамидная смола содержит составляющие звенья дикарбоновой кислоты, включающие в себя звенья адипиновой кислоты, и составляющие звенья диамина, включающие в себя звенья пентаметилендиамина и звенья гексаметилендиамина. Массовое отношение количества звеньев пентаметилендиамина к количеству звеньев гексаметилендиамина находится в пределах от 95:5 до 5:95. Звенья пентаметилендиамина образованы из пентаметилендиамина, полученного из лизина с использованием лизиндекарбоксилазы, способных продуцировать лизиндекарбоксилазу клеток, или продукта переработки таких клеток. По второму варианту полиамидная смола содержит составляющие звенья диамина при массовом отношение количества звеньев пентаметилендиамина к количеству звеньев гексаметилендиамина в пределах от 95:5 до 60: ...

СПОСОБ ПОЛУЧЕНИЯ АКРИЛАМИДА

Изобретение относится к получению акриламида. Способ осуществляют за счет применения биокатализатора для получения акриламида непрерывно из акрилонитрила, причем акриламид вводят в реактор, в который непосредственно подан акрилонитриловый раствор и инициируют непрерывные реакции привнесением акрилонитрила в контакт с биокатализатором. Технический результат - получение акриламида с низкими затратами с упрощенными процедурами, при непрерывном извлечении акриламидного раствора с требуемым уровнем концентрации. 4 з.п. ф-лы, 2 табл., 5 пр.

ФРАГМЕНТ НУКЛЕИНОВОЙ КИСЛОТЫ, КОДИРУЮЩИЙ ПОЛИПЕПТИД, ОБЛАДАЮЩИЙ АКТИВНОСТЬЮ НИТРИЛГИДРОТАЗЫ, РЕКОМБИНАНТНАЯ ПЛАЗМИДНАЯ ДНК PPCL 4, КОДИРУЮЩАЯ ПОЛИПЕПТИД, ОБЛАДАЮЩИЙ АКТИВНОСТЬЮ НИТРИЛГИДРАТАЗЫ, ШТАММ БАКТЕРИЙ ESCHERICHIA COLI - ПРОДУЦЕНТ ПОЛИПЕПТИДА, ОБЛАДАЮЩЕГО АКТИВНОСТЬЮ НИТРИЛГИДРАТАЗЫ, СПОСОБ ПОЛУЧЕНИЯ НИТРИЛГИДРАТАЗЫ

Изобретение относится к биотехнологии и генетической инженерии, в частности к получению ферментов клетчатого метаболизма. Сущность изобретения состоит в том, что из Pseudomonas chlororaphis В23 получен фрагмент ДНК, который кодирует полипептид, обладающий активностью нитрилгидратазы и способный гидратировать нитрилы в амиды. Получена рекомбинантная ДНК, содержащая этот ген, и трансформант, образованный путем трансформации штамма E. coli этой рекомбинантной ДНК. Изобретение, кроме того, включает способ получения нитрилгидратазы, используя трансформант. 4 с.п. ф-лы, 10 ил.

СПОСОБ ПОЛУЧЕНИЯ D-ПАНТОТЕНОВОЙ КИСЛОТЫ И/ИЛИ ЕЕ СОЛЕЙ, ИСПОЛЬЗУЕМЫХ В КАЧЕСТВЕ ДОБАВОК ПРИ ПРОИЗВОДСТВЕ КОРМОВ ДЛЯ ЖИВОТНЫХ

... 1. Способ получения D-пантотеновой кислоты и/или ее солей, отличающийся тем, что a) используют, по меньшей мере, один организм, продуцирующий D-пантотеновую кислоту, в котором нарушена регуляция биосинтеза пантотеновой кислоты-(pan) и/или изолейцина/валина-(ilv), и который в процессе ферментации в культуральной среде производит, по меньшей мере, 2 г/л соли D-пантотеновой кислоты, причем в культуральную среду добавляют 0-20 г/л свободного β-аланина и/или солей β-аланина, b) к образовавшемуся D-пантотенату добавляют соли, содержащие многовалентные катионы, в результате чего образуются многовалентные соли D-пантотеновой кислоты, c) раствор, содержащий многовалентные соли D-пантотеновой кислоты подвергают нанофильтрации, в результате чего концентрация многовалентных солей D-пантотеновой кислоты увеличивается, и d) ретентат, полученный после нанофильтрации, содержащий многовалентные соли D-пантотеновой кислоты подвергают затем сушке и/или формовке. 2. Способ по п.1, отличающийся тем, что в культуральную ...

ШТАММ RHODOCOCCUS RHODOCHROUS NCIMB 41164 И ЕГО ПРИМЕНЕНИЕ В КАЧЕСТВЕ ПРОДУЦЕНТА НИТРИЛГИДРАТАЗЫ

... 1. Микроорганизм, представляющий собой штамм Rhodococcus rhodochrous NCIMB 41164 или его мутант. 2. Способ культивирования штамма микроорганизма Rhodococcus rhodochrous NCIMB 41164 или его мутанта в культуральной среде, содержащей мочевину или производное мочевины. 3. Способ по п.2, в котором мочевину или производное мочевины вносят в культуральную среду по меньшей мере через 6 ч после начала выращивания микроорганизма. 4. Способ по п.2, в котором культуральная среда содержит менее 0,2 г/л мочевины или производного мочевины в течение по меньшей мере первых 6 ч культивирования микроорганизма, после чего в культуральную среду добавляют мочевину или производное мочевины. 5. Способ по п.2, в котором культуральная среда содержит менее 0,2 г/л мочевины или производного мочевины в течение по меньшей мере первых 12 ч культивирования микроорганизма, после чего в культуральную среду добавляют мочевину или производное мочевины. 6. Способ по п.2, в котором мочевину или производное мочевины добавляют ...

СПОСОБ ПРОДУЦИРОВАНИЯ ОРГАНИЧЕСКИ СВЯЗАННОГО ВИТАМИНА В

... 1. Способ продуцирования органически связанного витамина В, предпочтительно выбираемого из витамина B, В, В, В, В, В, В, Вили их смесей, в растениях, в котором всхожие семена растений сначала замачиваются в растворе соответствующего витамина или смеси витаминов и в процессе прорастания опрыскиваются растворами, содержащими соответствующий витамин.2. Способ по п.1, отличающийся тем, что семена растения проращиваются путем их предварительного замачивания в растворе соответствующего витамина и вследствие последующего опрыскивания процесс прорастания заканчивается через несколько дней.3. Способ по п.1, отличающийся тем, что семена растения пропитываются раствором в течение от 1 до 24 ч, предпочтительно в течение максимум 16 ч.4. Способ по п.1, отличающийся тем, что массовое отношение семян растения к содержащему соответствующий витамин раствору для их пропитки составляет от 1:1 до 1:20, предпочтительно до 1:10, наиболее предпочтительно до 1:5.5. Способ по п.1, отличающийся тем, что количество ...

СПОСОБ ПОЛУЧЕНИЯ АЛКИЛ(МЕТ)АКРИЛАТОВ С ИСПОЛЬЗОВАНИЕМ ФЕРМЕНТАТИВНОГО ГИДРОЛИЗА ЦИАНГИДРИНА

... 1. Способ получения алкил(мет)акрилатов, отличающийся тем, что способ включает в себя одну стадию, на которой циангидрин подвергается гидролизу с помощью фермента, остаточная активность которого после превращения метакрилнитрила в присутствии 20 мМ ионов цианида при температуре 20°С по истечении 30 мин составляет, по меньшей мере, 90% остаточной активности фермента, который при прочих одинаковых условиях использовался в отсутствие ионов цианида. ! 2. Способ по п.1, отличающийся тем, что остаточная активность после превращения в присутствии 50 мМ ионов цианида составляет, по меньшей мере, 60%. ! 3. Способ по п.1, отличающийся тем, что используют микроорганизмы, производящие и содержащие фермент, или их лизат. ! 4. Способ по п.3, отличающийся тем, что используется неподвижная клетка микроорганизма. ! 5. Способ по п.1, отличающийся тем, что используется очищенный фермент. ! 6. Способ по п.1, отличающийся тем, что фермент происходит из микроорганизмов рода Pseudomonas. ! 7. Способ по п.6, отличающийся ...

СПОСОБ ПОЛУЧЕНИЯ D-ПАНТОТЕНОВОЙ КИСЛОТЫ И/ИЛИ ЕЕ СОЛЕЙ, ИСПОЛЬЗУЕМЫХ В КАЧЕСТВЕ ДОБАВОК ПРИ ПРОИЗВОДСТВЕ КОРМОВ ДЛЯ ЖИВОТНЫХ

... 1. Способ получения D-пантотеновой кислоты и/или ее солей, отличающийся тем, что a) используют, по меньшей мере, один организм, продуцирующий D-пантотеновую кислоту, в котором нарушена регуляция биосинтеза пантотеновой кислоты-(pan) и/или изолейцина/валина-(ilv), и который в процессе ферментации в культуральной среде производит, по меньшей мере, 2 г/л соли D-пантотеновой кислоты, причем в культуральную среду добавляют 0-20 г/л свободного β-аланина и/или солей β-аланина, b) ферментационный раствор, содержащий D-пантотенат, пропускают через колонку с анионообменной смолой, c) связанный с анионообменной смолой D-пантотенат элюируют в виде свободной D-пантотеновой кислоты раствором, содержащим неорганические или органические соли кальция и/или магния в виде D-пантотената кальция и/или магния, или раствором соляной кислоты, d) при необходимости, рН элюата, содержащего свободную D-пантотеновою кислоту, добавлением кальциевых и/или магниевых оснований доводят до значения 3-10, при этом получают ...

СПОСОБ ПОЛУЧЕНИЯ СОЕДИНЕНИЯ ПРИ РЕГУЛИРОВАНИИ ТЕМПЕРАТУРЫ РЕАКЦИИ И ИСПОЛЬЗОВАНИИ БИОКАТАЛИЗАТОРА

... 1. Способ непрерывного получения соединения с использованием биокатализатора в двух или множестве реакторов, в котором более позднюю температуру реакции устанавливают выше более ранней температуры реакции между реакторами. 2. Способ получения по п.1, отличающийся тем, что более позднюю температуру реакции устанавливают выше более ранней температуры реакции, по меньшей мере, на 1єC между реакторами. 3. Способ получения по п.1, отличающийся тем, что более позднюю температуру реакции устанавливают выше более ранней температуры реакции, по меньшей мере, на 5єC между реакторами. 4. Способ получения соединения по любому из пп.1-3, отличающийся тем, что биокатализатором являются микробные клетки или продукт их переработки. 5. Способ получения соединения по п.4, отличающийся тем, что получаемым соединением является амидное соединение. 6. Способ получения соединения по п.5, отличающийся тем, что получаемым соединением является акриламид, никотинамид или 5-циановалерамид. 7. Способ получения соединения ...

CПOCOБ ПOЛУЧEHИЯ AMИДOB

Способ непрерывного получения водных растворов акриламида или метакриламида

Изобретение касается производных ненасыщенных кислот, в частности способа непрерывного получения водных растворов акриламида и метакриламида, используемых в производстве полимеров. Цель - упрощение процесса. Синтез ведут гидролизом соответствующего нитрила в водной среде при PH 7 - 9, 0 - 10°С, скорости истока каждого реагента в реакторе 0,4 - 4 ч-1в присутствии штаммов микроорганизмов N=771 или N=774 вида CORYNEBACTERIUM или штамма N 775 вида NOCORDIA (депонированы в Институте ферментных исследований Агентства промышленной науки и техники Министерства мировой торговли и промышленности Японии, г.Ибараги). Исходную смесь предварительно разбавляют частью продукта, отводимого из реактора, при массовом соотношении 1:(1 - 9). В этих условиях достигается снижение количества используемых колонн в процессе с 8 до 1.

Способ получения амида

Изобретение относится к биотехнологии и может быть использовано в производстве амида микробиологическим способом. Цель изобретения - ускорение способа. Амид получают путем гидратации нитрильного соединения с помощью суспензии грамположительных микроорганизмов при 0-20°С и PH среды 6-9, при этом суспензию предварительно облучают светом либо процесс гидратации ведут при освещении, либо осуществляют предварительную обработку светом суспензии и производят таковую по ходу реакции. Для этого используют световую энергию в количестве не менее 1.10-2мкЕ/г клеток в 1 с. с длиной волны 200-800 нм. Реакцию гидратации проводят в сосуде, состоящем по меньшей мере частично из материала, не пропускающего свет. 3 з.п.ф-лы, 6 табл.

DNA-SEQUENZ, DIE EIN POLYPEPTID MIT NITRILHYDRATASE-AKTIVITAET CODIERT, UND VERFAHREN ZUR HERSTELLUNG VON AMIDEN AUS NITRILEN MIT EINER TRANSFORMANTE, DIE DIE DNA-SEQUENZ ENTHAELT

Novel nannocheline and methyl ester(s) - active against Gram positive microorganisms

Nannocheline and its Me esters of formula (I) and their nontoxic salts are new. (In (I), R and R' are each H or Me. Prepn. of cpds. (I) comprises propagation of nannocystis exedens (DSM 5530) in a suitable culture medium in the presence of an anion ion exchange resin; then sepn. of the resin, elution with MeOH, and purification of the active fraction by chromatographic sepn. on sephadex and then silica gel. USE - (I) are new active against Gram positive microorganisms, but incompletely so against Escherichia coli and Saccharomyces cerevisiae.

Neue Phthalaldehyd-Derivate, Verfahren zu deren Herstellung und deren Verwendung

METHOD FOR PURIFYING REACTION SOLUTION OBTAINED BY USING MICROBIAL CELL, IMMOBILIZED MICROBIAL CELL, OR IMMOBILIZED ENZYME

BIOTECHNOLOGISCHE HERSTELLUNG VON POLYGLUTAMINSÄURE

ENZYMATISCHES VERFAHREN ZUR STEREOSELEKTIVEN HERSTELLUNG VON THERAPEUTISCHEN AMIDEN

Verfahren zur Herstellung enantiomerenangereicherter Amine und Amide durch enzymatische Racematspaltung

Die vorliegende Erfindung betrifft ein Verfahren zur Herstellung enantiomerenangereicherter Amine durch Spaltung eines racemischen Gemisches eines Reaktionsproduktes aus einem chiralen Amin und einem Acyldonor oder Umsetzung eines Amins mit einem Acyldonor in Gegenwart einer Hydrolase und anschließende Trennung des enantiomerenangereicherten Amids vom enantiomerenangereicherten Amin.

HERSTELLUNG VON AMIDEN

Aflatoxin production inhibitor, and method for controlling aflatoxin contamination using the same

Pantolactone hydrolase

PROCESS FOR PRODUCING ACRYLAMIDE OR METHACYLAMIDE UTILIZING MICRO-ORGANISMS

DIFLUOROBENZAMIDE

Compounds and pharmaceutically acceptable salts thereof

AI-77 compounds, pharmaceutically acceptable salts thereof, and a process for the preparation thereof are described, said compounds having the formulae (I) (II) wherein: X is NR6 or O; Y is NHR5 or combine with Z to provide a link for bonding C and C; Z is H or combines with Y to provide a link for bonding C and C; R1, R3 and R5 are each H, R', -CH2R, or -COR; R6 is H or R; R7 is H, R or CH2R; R is a hydrocarbon group consisting of a saturated or unsaturated straight or branched aliphatic group of C1 to C17, an aromatic group of C6 to C10, a cage type group of C7 to C10, a monocyclic aliphatic group of C3 to C8, an aromatic-aliphatic group of C7 to C15, a heterocyclic hydrocarbon of C1 to C9, wherein the above hydrocarbons can be substituted with one or more groups selected from halogen, oxo, carboxyl, hydroxyl, a saturated or unsaturated straight or branched aliphatic group of C1 to C5, an aromatic group of C6 to C10, a monocyclic aliphatic group of C3 to C8, an aromatic-aliphatic group ...

Preparation of pregabalin and related compounds

Process for preparing enantiomerically enriched N-derivatised lactams.

The present invention relates to a process for the production of substantially enantiomcrically pure intermediates of formula (IV), wherein P is an activating and protecting group, from their racemates by treating the mixture with an acylasc enzyme derived from Bacillus sp..

Preparation of pregabalin and related compounds

Process for preparing enantiomerically enriched N-derivatised lactams

Preparation of pregabalin and related compounds

Preparation of pregabalin and related compounds

Process for preparing enantiomerically enriched N-derivatised lactams

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

AMINO-ALCOHOL DERIVATIVES

PROCEDURE FOR THE PRODUCTION OF NEW PHOSPHONIC ACIDS

PROCEDURE FOR THE PRODUCTION OF D-PANTOTHENSÄURE ONES AND/OR THEIR SALTS AS ADDITIVE FOR ANIMAL ANIMAL FEEDS

CITRULLIMYCINE, PROCEDURE FOR YOUR PRODUCTION AND YOUR USE AS DRUGS

MANUFACTURING PROCESS FOR OPTICALLY ACTIVE CHIRALE OF AMINES

PROCEDURE FOR THE PRODUCTION OF THEANIN

PROCEDURE FOR THE PRODUCTION OF CHIRALEN ALCOHOLS

VERFAHREN ZUR HERSTELLUNG VON CHIRALEN ALKOHOLEN

The invention relates to a method for producing an enantiopure alcohol of general formula (Ia) or (Ib), wherein R1, R2, R3, R4, R5 and R6 each represent hydrogen, halogen, a C1-C6 alkyl or C1-C6 alkoxy group, with the proviso that at least one of the groups R1, R2, R3, R4, R5 and R6 is different from the remaining five groups and with the additional proviso that at least one of the groups R1, R2, R3, R4, R5 and R6 is a halogen. The invention is characterized in that a ketone of general formula (II), wherein R1, R2, R3, R4, R5 and R6 are defined as above, is enzymatically reduced in the presence of an S-specific or R-specific dehydrogenase/oxidoreductase using NADH or NADPH as the cofactor.

ENZYMATIC PROCEDURES FOR the PRODUCTION 4SUBSTITUIERTER 3-HYDROXYBUTTERSÄURE-DERIVATE

VERFAHREN ZUR PRODUKTION VON ORGANISCH GEBUNDENEM VITAMIN B

PROCEDURE FOR THE PRODUCTION OPTICALLY OF AN ENRICHMENT OF TERTIARY ALCOHOL

VERFAHREN ZUR STEREOSPEZIFISCHEN HERSTELLUNG VON D-CARBAMYLDERIVATEN VON ALPHA-HYDROXYSAEUREN

VERFAHREN ZUM HERSTELLEN DES NEUEN ANTIBIOTIKUMS BMG162-AF2 ODER EINES SEINER PHARMAZEUTISCH ANNEHMBARER SALZE

TRUNK OF RHODOCOCCUS RHODOCHROUS NCIMB 41164 AS WELL AS ITS USE AS NITRILHYDRATASEPRODUZENT

CYANIDE-TOLERANT NITRILHYDRATASEN

CHEMICAL-ENZYMATIC PEPTIDSYNTHESE OF MEANS OF CTERMINALER ESTER TRANSFORMATION

PROCEDURE FOR THE PRODUCTION OF AMINO ACID DERIVATIVES FROM HYDROXYIMINOSÄUREN

PROCEDURE AND MICROORGANISM FOR THE IMPROVED PRODUCTION OF PANTOTHENAT

PROCEDURE FOR THE STEREOSPEZIFI PRODUCTION OF D-CARBAMYLDERIVATEN OF ALPHA HYDROXYSAEUREN

PROCEDURE FOR THE PRODUCTION OF A WITH B VITAMINS ENRICHMENT PLANT

PROCEDURE FOR THE PRODUCTION OF NEW PHOSPHON SAEUREN

PROCEDURE FOR THE PRODUCTION OF ORGANIC CHEMICAL COMPOUNDS.

PROCEDURE FOR THE REDUCTION OF KETONEN.

ANTI-PARASITIC MATERIALS AND PROCESSES TO YOUR PRODUCTION AS WELL AS ANTI-PARASITIC EFFECTIVE PREPARATIONS, IN PARTICULAR ANTI-MALARIA MEANS, WHICH THESE MATERIALS CONTAINING.

PROCEDURE FOR the PRODUCTION OF 2-HYDROXYMETHYLPYROLIDIN-3,4-DIOLEN

PROCEDURE FOR THE PRODUCTION OF D-PANTOTHENSÄURE ONES AND/OR THEIR SALTS AS ADDITIVE FOR ANIMAL ANIMAL FEEDS

PROCEDURE FOR THE PRODUCTION OF OPTICALLY ACTIVE ONE AMINOALKOHOLEN THROUGH RACEMATSPALTUNG

SEPARATION FROM CHIRALEN AMINES

TRANSFORMANT CAPABLE OF the PRODUCTION OF D AMINOSÄUREOXIDASEN

D-PANTOTHENSÄURE ONE AND/OR ONE OF THEIR SALTS ABSTENTIONS OF ANIMAL FEED ADDITIVES AND PROCEDURES FOR THEIR PRODUCTION

PROCEDURE FOR THE CLEANING OF VIRUSES WITH CHROMATOGRAPHY

ENZYMATIC SYNTHESIS OPTICAL OF ACTIVE HYDROXAMIC ACIDS AND YOUR CONVERSION TO OPTICAL ACTIVE ONE PRIMARY AMINES BY LOT EN RELOCATION

PROCEDURE FOR THE PRODUCTION OF DIHYDROXYPYRIMIDIN DERIVATIVES

PRODUCTION OF D-PANTOINSÄURE ONES AND D PANTOTHENSÄURE

ENZYMATIC PRODUCTION OF PHOSPHOLIPIDEN IN AQUEOUS MEDIA

PROCEDURE FOR THE PRODUCTION OF AMIDES

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

PROCEDURE FOR THE PRODUCTION OF AN AQUEOUS ACRYLAMIDE SOLUTION WITH A BIOCATALYST

Process for producing a polyacrylamide solution with increased viscosity

The present invention relates to a method for producing a polyacrylamide solution having increased viscosity. In particular, the present invention is related to the separation of a biocatalyst from an aqueous acrylamide solution prepared utilizing the biocatalyst prior to polymerization of the aqueous acrylamide solution to polyacrylamide. A polyacrylamide solution having increased viscosity is well suited to be used in tertiary oil recovery. Accordingly, the present application provides means and methods to crucially improve the quality of polyacrylamide solutions for use in tertiary oil recovery.

Enzymatic preparation of phospholipids in aqueous media

Enzyme-catalyzed racemic cleavage of primary amines

Process for producing polymers

Transformant producing secondary metabolite modified with functional group and novel biosynthesis genes

A process for carrying out enzymatically catalyzed conversions of organic compounds

Physiologically active substances tkr2449, process for producing the same, and microorganism

Preparation of alpha-ketopimelic acid

The present invention relates to a method for preparing alpha-ketopimelic acid, comprising converting 2-hydroxyheptanedioic acid into alpha-ketopimelic acid, which conversion is catalysed using a biocatalyst. Further, the invention relates to a heterologous cell, comprising a nucleic acid sequence encoding an enzyme having catalytic activity in the conversion of 2-hydroxyheptanedioic acid into alpha-ketopimelic acid. Further, the invention relates to the use of a heterologous cell according to the invention in the preparation of caprolactam, diaminohexane or adipic acid.

Methods of producing carbamoyl phosphate and urea

The present invention relates to a method of producing carbamoyl phosphate, the method comprising reacting ammonia, ATP, bicarbonate and CO ...

Process for the preparation of benzoxazine derivatives and intermediates therefor

Method of manufacturing compound with biocatalyst by using controlled reaction temperature

Process for preparing enantiomerically enriched N-derivatised lactams

Mutants which produce a potentiator of bacillus pesticidal activity

ENATIOSELECTIVE REDUCTION OF KETONES BY MICROORGANISMS TO FORM SECONDARY ALCOHOLS IN PARTICULAR FOR SYNTHESIS OF RENIN INHIBITORS

Physiologically active substances tkr1785's, process for the preparation thereof, and microbe

Process for producing amide compounds

Preparation of adipic acid

The invention relates to a method for preparing adipic acid, comprising converting alpha-ketoglutaric acid (AKG) into alpha-ketoadipic acid (AKA), converting alpha-ketoadipic acid into alpha-ketopimelic acid (AKP), converting alpha-ketopimelic acid into 5-formylpentanoic acid (5-FVA), and converting 5-formylpentanoic acid into adipic acid, wherein at least one of these conversions is carried out using a heterologous biocatalyst.The invention further relates to a heterologous cell, comprising one or more heterologous nucleic acid sequences encoding one or more heterologous enzymes capable of catalysing at least one reaction step in said method.

Methods of increasing dihydroxy acid dehydratase activity to improve production of fuels, chemicals, and amino acids

The present invention is directed to recombinant microorganisms comprising one or more dihydroxyacid dehydratase (DHAD)-requiring biosynthetic pathways and methods of using said recombinant microorganisms to produce beneficial metabolites derived from said DHAD-requiring biosynthetic pathways. In various aspects of the invention, the recombinant microorganisms may be engineered to overexpress one or more polynucleotides encoding one or more Aft proteins or homologs thereof. In some embodiments, the recombinant microorganisms may comprise a cytosolically localized DHAD enzyme. In additional embodiments, the recombinant microorganisms may comprise a mitochondrially localized DHAD enzyme. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces Glade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Preparation of alpha-ketopimelic acid

The invention relates to a method for preparing alpha-ketopimelic acid, comprising converting alpha-ketoglutaric acid into alpha-ketoadipic acid and converting alpha-ketoadipic acid into alpha-ketopimelic acid, wherein at least one of these conversions is carried out using a heterologous biocatalyst. The invention further relates to a heterologous cell, comprising one or more heterologous nucleic acid sequences encoding one or more heterologous enzymes capable of catalysing at least one reaction step in the preparation of alpha-ketopimelic acid from alpha-ketoglutaric acid.

Aflatoxin production inhibitor and method for controlling aflatoxin contamination using the same

The present invention relates to provide an aflatoxin production inhibitor that inhibits aflatoxin production specifically and efficiently, is highly safe, and is practical, and an efficient production method thereof; and a method for controlling aflatoxin contamination that uses the aflatoxin production inhibitor, specifically relating to an aflatoxin production inhibitor that includes at least one of a dioctatin represented by the following formula (I) and a derivative thereof, as an active ingredient: where, in the formula (I), R represents one of hydrogen and a methyl group.

Process for preparing a carbamate compound

There is provided a process for preparing a carbamate compound, which is easy and commercially advantageous in that a carbamate compound can be produced with high yield from an amine compound and a carbonate compound. A process for preparing a carbamate compound which comprises the step of reacting an amine compound which has at least one amino group per molecule wherein the amine compound is selected from the group consisting of an aliphatic amine which may be substituted by an alicyclic group or an aromatic group or which may be interrupted by an alicyclic group or an aromatic group, and an alicyclic amine which may be substituted by an aliphatic group, with a carbonate compound in the presence of at least one organic solvent selected from the group consisting of a saturated cyclic hydrocarbon, an unsaturated cyclic hydrocarbon, and a non-cyclic ether by using a hydrolase.

Novel amidase, gene for the same, vector, transformant, and method for production of optically active carboxylic acid amide and optically active carboxylic acid by using any one of those items

The present invention has its object to provide a novel polypeptide having amidase activity to selectively hydrolyze S-enantiomer in racemic nipecotamide, a DNA encoding the polypeptide, a vector containing the DNA, a transformant transformed with the vector, and a method for producing an optically active carboxylic acid amide and an optically active carboxylic acid in which a racemic carboxylic acid amide is hydrolyzed with the polypeptide or the transformant.

Process for Producing Aliskiren

A new route of synthesis of the compound Aliskiren of formula (I), used in the treatment of hypertension, is described.

Host Cells and Methods for Producing Cinnamoyl Anthranilate and Analogs Thereof

The present invention provides for a method of producing a cinnamoyl anthranilate, or analog thereof, in a genetically modified host cell. 1. A genetically modified host cell comprising an hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT , EC 2.3.1.144) , or functional fragment thereof , capable of catalyzing the formation of a cinnamoyl anthranilate , or analog thereof , from a cinnamoyl-CoA , or analog thereof , and an anthranilate , or analog thereof.2Dianthus caryophyllus. The host cell of claim 1 , wherein the HCBT has an amino acid sequence at least 70% identical to the amino acid sequence of HCBT.3Dianthus caryophyllus. The host cell of claim 2 , wherein the HCBT is a HCBT.4. The host cell of further comprising a 4-coumarate:CoA ligase (4CL claim 1 , EC 6.2.1.12) claim 1 , or functional fragment thereof claim 1 , capable of catalyzing the formation of a cinnamic acid claim 1 , or analog thereof claim 1 , into a corresponding cinnamoyl-CoA thioester claim 1 , or analog thereof.5Arabidopsis thaliana.. The host cell of claim 1 , wherein the 4CL has an amino acid sequence at least 70% identical to the amino acid sequence of 4-coumarate:CoA ligase 5 (4CL5) of6Arabidopsis thaliana.. The host cell of claim 5 , wherein the 4CL is a 4-coumarate:CoA ligase 5 (4CL5) of7. The host cell of claim 4 , wherein the cinnamic acid claim 4 , or analog thereof claim 4 , is a hydroxycinnamic acid claim 4 , and the host cell lacks any enzyme capable of catalyzing the decarboxylation of a hydroxycinnamic acid.8. The host cell of claim 7 , wherein the host cell is a yeast cell and the pad1 gene is deleted or has a reduction of expression claim 7 , or the host cell has a reduced capability to catabolize claim 7 , metabolize claim 7 , or modify the hydroxycinnamic acid claim 7 , or analog thereof.10. A method of producing a cinnamoyl anthranilate claim 1 , or analog thereof claim 1 , comprising: culturing the genetically modified host cell of under ...

ENZYMATIC RESOLUTION OF RACEMIC (2R,S)-2-(ACETYLAMINO)-3-METHOXY-N-(PHENYLMETHYL)PROPANAMIDE

The present invention is concerned with a process of preparing (R)-lacosamide. The process comprises providing an (R,S)-lacosamide precursor and contacting the same with at least an enzyme in the presence of a solvent. The enzyme either stereoselectively hydrolyzes or acetylates an (R)- or (S)-enantiomer of the (R,S)-lacosamide precursor. The process further comprises where appropriate also concurrently, or successively, employing one or more reagents capable of converting the hydrolysed or acetylated (R)- or (S)-enantiomer to (R)-lacosamide. 140.-. (canceled)42. The process of claim 41 , wherein step (ii) comprises contacting the (R claim 41 ,S)-compound of formula (II) with one or more enzymes in the presence of a solvent and optionally an acetyl donor compound to form an enantiomerically enriched or enantiomerically pure (R)-enantiomer of the compound formula (II) wherein Ris CHC(═O)— claim 41 ,{'sub': '1', 'wherein the one or more enzymes is capable of stereoselectively hydrolyzing the (R)- or (S)-enantiomer of a compound of formula (II) when the solvent is a protic solvent or is capable of stereoselectively acetylating the (R)-enantiomer of a compound of formula (II) when Ris a first intermediate moiety and when an acetyl donor compound is present, and'}{'sub': '2', 'wherein step (iii) comprises optionally further comprising contacting the (R,S)-compound of formula (II) or the enriched or enantiomerically pure (R)-enantiomer of the compound formula (II) with one or more reagents capable of converting a second intermediate moiety into —NHCHPh.'}45. The process of claim 41 , wherein the enzyme capable of stereoselectively acetylating either the (R)- or (S)-enantiomer of a compound of formula (II) is a lipase.46Candida antarcticaCandida antarcticaPseudomonas cepacia. The process of claim 45 , wherein the lipase is selected from the group consisting of lipase A (CAL-A) claim 45 , lipase B (CAL-B) claim 45 , and lipase.47. The process of claim 43 , wherein the ...

CHEMICAL AND BIOLOGICAL AGENTS FOR THE CONTROL OF MOLLUSCS

Compositions and methods for controlling molluscs, members of the Gastropoda and Bivalvia classes which includes but is not limited to lactones, lactams, carbamates, amides, and/or carboxylic acid containing compounds as active ingredients and/or compounds derived from and/or . Also provided are methods and compositions for increasing the efficacy of chemical and biological control for invasive molluscs in open waters, power plants, and drinking water treatment facilities under coldwater conditions. 1. A combination comprising at least one or more substances effective for controlling a member of a Gastropoda and Bivalvia class and an inert material.2PseudomonasPseudomonas. The combination according to claim 1 , wherein said substances are derived from a species or cell suspension or toxin derived from a species.3Pseudomonas protegensPseudomonas protegens.. The combination according to claim 1 , wherein said substances are derived from a or cell suspension derived from4PseudomonasPseudomonas. The combination according to claim 1 , wherein said substances are derived from a strain claim 1 , having the identifying characteristics of ATCC 55799.5Pseudomonas. The combination according to claim 1 , wherein the composition comprises a substance that is a cell suspension comprising cells having the toxin producing characteristics of ATCC 55799.6. The combination of claim 1 , wherein said combination is a composition.7ErwiniaPseudomonas. A method for controlling one or more molluscs in a location where control is desired comprising introducing into said location at least one of (a) a cell suspension or extract derived from sp. Cells; (b) one or more compounds claim 1 , wherein said compounds are lactone claim 1 , lactam claim 1 , carbamate claim 1 , carboxylic acid and/or amide compounds or composition comprising said compounds claim 1 , with the proviso that said compounds are not gamma-octalactone claim 1 , gamma-nonalactone claim 1 , gamma-decanolactone claim 1 , gamma- ...

Method for producing gardenia blue pigment

Provided is a method for producing a gardenia blue pigment resistant to discoloration which may occur in colored sugar-coated food or pharmaceutical products. The method for producing such a gardenia blue pigment comprises performing membrane separation using a membrane (for example, an ultrafiltration membrane) with a molecular weight cut-off of 3000 Da or larger for removal of low-molecular compounds from a solution resulting from β-glucosidase treatment of an iridoid glycoside (for example, geniposide etc.) in the presence of a protein hydrolysate (for example, a casein protein hydrolysate), the iridoid glycoside being obtainable by extraction from fruits of Gardenia jasminoides (Rubiaceae). Also provided is a sugar-coated food or pharmaceutical product (for example, a sugar-coated tablet, a sugar-coated chewing gum, etc.) having a sugar-coating layer colored with the gardenia blue pigment obtainable by the above-described method.

IMMOBILIZATION METHOD OF BIOACTIVE MOLECULES USING POLYPHENOL OXIDASE

A method is provided for immobilizing a bioactive molecule onto a surface using polyphenol oxidase. In the presence of polyphenol oxidase, a bioactive molecule containing a phenol or catechol group can be simply in situ oxidized within a short time to dopa or dopaquinone which forms a coordinate bond with a metal or polymer substrate, thus immobilizing the bioactive molecule onto the surface with stability. Based on the surface immobilization of bioactive molecules using polyphenol oxidase, various bioactive molecules such as osteogenetic peptides and growth factors can be simply immobilized to medical metal or polymer substrate surfaces such as orthopedic or dental implants which can be then effectively used to induce rapid osteogenesis after being transplanted. Also, antithrombotic agents and/or entothelialization inducing agents may be immobilized to medical substrates for vascular systems, such as stents and artificial blood vessels, thus guaranteeing hemocompatibility to the medical substrates. 1. A method for immobilization of a bioactive molecule on a surface , comprising:preparing a phenol-, catechol- or its derivative containing bioactive molecule (step 1); and treating a substrate surface with the bioactive molecule in presence of polyphenol oxidase to immobilize the bioactive molecule onto the surface (step 2).2. The method according to claim 1 , wherein the bioactive molecule contains a cell adhesion peptide containing a tyrosine.3. The method according to claim 2 , wherein the cell adhesion peptide is selected from the group consisting of peptides of SEQ ID NO. 1 (RGD-Y) claim 2 , SEQ ID NO. 2 (KQAGDV-Y) claim 2 , SEQ ID NO. 3 (YIGSR) claim 2 , SEQ ID NO. 4 (REDV-Y) claim 2 , SEQ ID NO. 5 (IKVAN-Y) claim 2 , SEQ ID NO. 6 (RNIAEIIKDI-Y) claim 2 , SEQ ID NO. 7 (KHIFSDDSSE-Y) claim 2 , SEQ ID NO. 8 (VPGIG-Y) claim 2 , SEQ ID NO. 9 (FHRRIKA-Y) claim 2 , SEQ ID NO. 10 (KRSR-Y) claim 2 , SEQ ID NO. 11 (NSPVNSKIPKACCVPTELSAI-Y) claim 2 , SEQ ID NO. 12 (APGL-Y) ...

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenyl-methyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 80% sequence identity to SEQ ID NO: 2 and one or more amino acid differences relative to SEQ ID NO: 2 selected from the following:X143C, E, F, G, H, K, M, P, Q, S, T, or W;X246A, E, G, I, L, N, P, S, T, or V;X277C, D, E, G, H, L, M, P, S, T, V, or W;X278A, C, G, H, K, N, Q, S, T, or V;X280L, T, or W;X281A, C, H, K, L, M, N, R, T, V, W, or Y;X326A, D, E, F, G, H, L, M, N, P, R, V, or W;X426G, Q, or T;X432E, I, K, N, Q, T, V, or W;X433 S;X435G, K, V, or Y;X490A, C, D, E, G, I, L, M, N, S, or Y; andX532M.2. The polypeptide of claim 1 , wherein the polypeptide is capable of converting the acid substrate compound (1b) to compound (2b) (R-enantiomer) or its opposite enantiomer compound (S-enantiomer) with at least 2-fold improved activity relative to the wild-type polypeptide of SEQ ID NO: 2.3. The polypeptide of claim 1 , wherein the amino acid sequence comprises one or more amino acid differences relative to SEQ ID NO: 2 selected from: X143G claim 1 , K claim 1 , M claim 1 , P claim 1 , Q claim 1 , S claim 1 , or W; X246A claim 1 , E claim 1 , G claim 1 , I claim 1 , L claim 1 , P claim 1 , S claim 1 , T claim 1 , or V; X278G claim 1 , or N; X426Q claim 1 , or T; X432E claim 1 , I claim 1 , L claim 1 , Q claim 1 , S claim 1 , T claim 1 , V claim 1 ...

Microorganisms and methods for the biosynthesis of adipate, hexamethylenediamine and 6-aminocaproic acid

The invention provides a non-naturally occurring microbial organism having a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in the respective 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The invention additionally provides a method for producing 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid. The method can include culturing a 6-aminocaproic acid, caprolactam or hexametheylenediamine producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway enzyme in a sufficient amount to produce the respective product, under conditions and for a sufficient period of time to produce 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid.

Process for the preparation of lacosamide

There is provided a process for the preparation of Lacosamide (which is a useful medicament) of formula I, which comprises an enantioselective enzymatic acylation.

Ketol-acid reductoisomerase enzymes and methods of use

Provided herein are polypeptides having ketol-aid reductoisomerase activity as well as microbial host cells comprising such polypeptides. Polypeptides provided herein may be used in biosynthetic pathways, including, but not limited to, isobutanol biosynthetic pathways.

METHOD FOR PRODUCING AQUEOUS ACRYLAMIDE SOLUTION

There is provided a method for producing an aqueous acrylamide solution by reacting a composition including acrylonitrile with water to produce acrylamide, in which the composition including acrylonitrile includes 3 to 15 mg of propionitrile per 1 kg of the total weight of the composition including acrylonitrile. According to the present invention, a production method with which it is possible to suppress acrylamide polymerization without lowering quality and thereby obtain a stable aqueous acrylamide solution can be provided. 1. A method for producing an aqueous acrylamide solution , the method comprising reacting a composition comprising acrylonitrile and water to produce acrylamide , wherein the composition further comprises 3 to 15 mg of propionitrile per 1 kg of the total weight of the composition.2. The method according to claim 1 , wherein the reacting occurs in the presence of a biocatalyst.3. The method according to claim 1 , wherein the composition further comprises 2 to 20 mg/kg of acetonitrile per 1 kg of the total weight of the composition.4. The method according to claim 1 , wherein a concentration of the acrylamide in the aqueous acrylamide solution is 30 to 60% by mass relative to the total mass of the aqueous acrylamide solution.5. The method according to claim 2 , wherein the composition further comprises 2 to 20 mg/kg of acetonitrile per 1 kg of the total weight of the composition.6. The method according to claim 2 , wherein a concentration of the acrylamide in the aqueous acrylamide solution is 30 to 60% by mass relative to the total mass of the aqueous acrylamide solution.7. The method according to claim 3 , wherein a concentration of the acrylamide in the aqueous acrylamide solution is 30 to 60% by mass relative to the total mass of the aqueous acrylamide solution. The present invention relates to a method for producing an aqueous acrylamide solution.The present application claims priority to Japanese Patent Application No. 2011-112428, which ...

MICROORGANISMS AND METHODS FOR PRODUCING ACRYLATE AND OTHER PRODUCTS FROM HOMOSERINE

This invention relates to microorganisms that convert a carbon source to acrylate or other desirable products using homoserine and 2-keto-4-hydroxybutyrate as intermediates. The invention provides genetically engineered microorganisms that carry out the conversion, as well as methods for producing acrylate by culturing the microorganisms. Also provided are microorganisms and methods for converting homoserine to 3-hydroxypropionyl-CoA, 3-hydroxypropionate (3HP), poly-3-hydroxypropionate and 1,3-propanediol. 1. A method for converting homoserine to 3-hydroxypropionyl-CoA comprising the steps of:{'smallcaps': L', 'L, 'a) converting homoserine to 2-keto-4-hydroxybutyrate, wherein this conversion is catalyzed by at least one enzyme selected from the group consisting of an aminotransferase, an -amino acid oxidase and an -amino acid dehydrogenase; and'}b) converting 2-keto-4-hydroxybutyrate to 3-hydroxypropionyl-CoA, wherein this conversion is catalyzed by at least one enzyme selected from the group consisting of a 2-ketoacid dehydrogenase and a combination of a 2-ketoacid decarboxylase and a dehydrogenase.2. The method of in which a recombinant microorganism overexpresses one or more genes to convert homoserine to 3-hydroxypropionyl-CoA.3. The method of in which the microorganism expresses a poly-3-hydroxyalkanoate synthase to further convert 3-hydroxypropionyl-CoA to a poly-3-hydroxyalkanoate containing 3-hydroxypropionate monomers.4. The method of further comprising the steps of:c) converting 3-hydroxypropionyl-CoA to acryloyl-CoA, wherein this conversion is catalyzed by a dehydratase; andd) converting acryloyl-CoA to acrylic acid, wherein this conversion is catalyzed by at least one enzyme selected from the group consisting of a thioesterase, a CoA-transferase, and a combination of a phosphate transferase and kinase.5. The method of in which a recombinant microorganism converts homoserine to acrylic acid.6. The method of in which 3-hydroxypropionyl-CoA is further ...

IN VIVO AND IN VITRO OLEFIN CYCLOPROPANATION CATALYZED BY HEME ENZYMES

The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention. 182-. (canceled)84. The reaction mixture of claim 83 , wherein each Rand Ris independently an optionally substituted Calkyl.85. The reaction mixture of claim 83 , wherein each Ris independently an optionally substituted Calkyl.86. The reaction mixture of claim 83 , wherein Ris a phenyl group.87. The reaction mixture of claim 83 , wherein the carbene precursor is a diazo reagent selected from the group consisting of diazomethane claim 83 , an α-diazoester claim 83 , an α-diazoamide claim 83 , an α-diazonitrile claim 83 , an α-diazoketone claim 83 , and an α-diazoaldehyde.89. The reaction mixture of claim 87 , wherein the diazo reagent is ethyl diazoacetate.90. The reaction mixture of claim 83 , wherein the olefinic substrate and the carbene precursor are part of the same substrate.91. The reaction mixture of claim 83 , wherein the cyclopropanation product is produced in vitro.92. The reaction mixture of claim 83 , wherein the reaction mixture further comprises a reducing agent.93. The reaction mixture of claim 83 , wherein the heme enzyme is localized within a whole cell and the cyclopropanation product is produced in vivo.94. The reaction mixture of claim 83 , wherein the cyclopropanation product is produced under anaerobic ...

Polypeptides Having Peroxygenase Activity and Polynucleotides Encoding Same

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polynucleotide encoding a non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO: 136 , and one or more amino acid substitutions compared to the naturally occurring polypeptide at one or more positions corresponding to positions in SEQ ID NO: 136 , selected from the group consisting of 75 , 79 , 82 , 99 , 110 , 166 , 172 , 208 , 216 , 273 , 324 , 364 , 395 , 412 , 491 , 503 , and 504.3. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 136 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO: 136.5. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 136 claim 1 , wherein said polypeptide further comprises one or more substitutions corresponding to substitutions in SEQ ID NO: ...

BIOCATALYSTS FOR THE PREPARATION OF HYDROXY SUBSTITUTED CARBAMATES

The present disclosure relates to engineered ketoreductase polypeptides for the preparation of hydroxyl substituted carbamate compounds, and polynucleotides, vectors, host cells, and methods of making and using the ketoreductase polypeptides. 2. The engineered ketoreductase polypeptide of claim 1 , wherein the amino acid sequence of said ketoreductase polypeptide comprises the substitution X40R claim 1 , and one or more residue differences as compared to SEQ ID NO:4 selected from: X7S; X17M; X17Q; X17R; X23V; X27L; X29G; X60I; X64V; X71P; X87L; X94A; X94P; X94S; X95M; X105G; X113I; X122A; X127R; X131S; X144V; X145L; X147I; X147L; X147Q; X150Y; X152G; X153G; X157C; X173L; X196M; X198S; X208R; X216R; X221S; X243S; X245I; X249F; X249G; and X249Y.3. The engineered ketoreductase polypeptide of claim 1 , wherein the amino acid sequence of said ketoreductase polypeptide comprises X40R claim 1 , and at least one or more residue differences as compared to SEQ ID NO:4 selected from: X17Q/R/M; X64V; X94P; X96L/Y; X144V; X147Q/UL; X157C; X195A/G; X196M; X199H; and X206L/F.4. The engineered ketoreductase polypeptide of claim 1 , wherein the ketoreductase polypeptide is capable of converting the substrate compound (2) to the product compound (1) with at least 10 fold the activity of the reference polypeptide of SEQ ID NO:4 claim 1 , wherein the amino acid sequence comprises the substitution X40R claim 1 , and one or more residue differences as compared to SEQ ID NO:4 selected from: X601; X71P; X94P; X94A; X95M; X96L; X96Y; X127R; X144V; X145I; X150Y; X152G; X153G; X157C; X195A; X195G; X196M; X198S; X199H; X206F/L claim 1 , X216R claim 1 , X2451 claim 1 , X245F; X249Y; and X249F.5. The engineered ketoreductase polypeptide of claim 1 , wherein the ketoreductase polypeptide has increased thermal stability as compared to the reference polypeptide of SEQ ID NO:4 or 32 claim 1 , wherein the amino acid sequence comprises the substitution X40R claim 1 , and one or more residue ...

Methods and Materials for Producing Hybrid Cell Lines

Methods and materials are described for producing immortalized hybrid cells composed of a fusion of immortalized multi-lineage progenitor cells (MLPC) and primary hepatocytes. The hybrid cells express the biological activity of the primary hepatocytes and the immortality and expansion capacities of the immortalized MLPC. The methods of culture and expansion of the resultant hybrid cells and the methods to confirm the characteristics of the hybrid cells are described. 1. A cell population comprising a plurality of hybrid cells , wherein each hybrid cell is composed of an immortalized multi-lineage progenitor cell (MLPC) and a primary somatic cell.2. The cell population of claim 1 , wherein said hybrid cell is created by the fusion of said immortalized MLPC and said primary somatic cell.3. The cell population of claim 1 , wherein said immortalized MLPC comprises a nucleic acid encoding a telomerase reverse transcriptase.4. The cell population of claim 1 , wherein said primary somatic cell is a hepatocyte.5. The cell population of claim 1 , wherein said hybrid cell has the biological activity associated with the primary somatic cell.6. The cell population of claim 1 , wherein the hybrid cell is immortalized and has the expandability of said immortalized MLPC.7. The cell population of claim 1 , wherein the hybrid cells can be expanded continuously in an expansion medium claim 1 , said expansion medium comprising hydrocortisone claim 1 , bovine serum albumin claim 1 , insulin claim 1 , transferrin claim 1 , selenium claim 1 , epithelial growth factor claim 1 , basic fibroblast growth factor claim 1 , fibroblast growth factor 4 claim 1 , hepatocyte growth factor claim 1 , stem cell factor claim 1 , oncostatin M claim 1 , bone morphogenic protein 4 claim 1 , and interleukin 1 beta.8. The cell population of claim 7 , the expansion medium further comprising an antibiotic.9. The cell population of claim 4 , wherein said fusion cells are positive for alkaline phosphatase claim ...

METHOD OF PRODUCING AMIDE COMPOUND

Provided is a method of producing an amide compound, the method including: obtaining a reaction solution containing an amide compound by bringing a microbial cell containing nitrile hydratase, or a processed product of the microbial cell, into contact with a nitrile compound in an aqueous medium in a first reactor; and causing the obtained reaction solution containing an amide compound to react in a second reactor having a plug-flow region, in which the Reynolds number in the second reactor is controlled to from 5 to 1,000. 1. A method of producing an amide compound , the method comprising:obtaining a reaction solution including an amide compound by bringing a microbial cell including nitrile hydratase, or a processed product of the microbial cell, into contact with a nitrile compound in an aqueous medium in a first reactor; andcausing the obtained reaction solution including an amide compound to react in a second reactor having a plug-flow region,a Reynolds number in the second reactor being controlled to from 5 to 1,000.2. The method of producing an amide compound according to claim 1 , wherein the Reynolds number in the second reactor is 10 or higher.3. The method of producing an amide compound according to claim 1 , wherein the Reynolds number in the second reactor is 100 or lower.4. The method of producing an amide compound according to claim 1 , wherein the second reactor comprises a tube reactor that has a tube diameter of 10 cm or more.5. The method of producing an amide compound according to claim 1 , wherein the Reynolds number in the second reactor is from 11 to 100.6. The method of producing an amide compound according to claim 1 , wherein the second reactor comprises a tube-type reactor that has a tube diameter of 12 cm or more. The present disclosure relates to a method of producing an amide compound.The hydration method using a nitrile compound as a raw material is used as one of the major process for producing an amide compound in many cases. ...

Regulation Method for Preparing Gamma-Polyglutamic Acid by Sludge Substrate Fermentation

A regulation method for preparing γ-polyglutamic acid by sludge substrate fermentation includes: 1) extraction of glutamic acid from sludge protein (high pressure hydrothermal treatment, gravity pressure filtration treatment), 2) secondary metabolic synthesis of γ-polyglutamic acid (activation of domesticated strains and secondary metabolic fermentation strains); and 3) preparation of pure γ-polyglutamic acid (acidification, centrifugation, filtration, precipitation based on polar repulsion, purification, impurity removal and drying). The present invention realizes a recycling of high-value carbon and nitrogen sources of sludge without secondary pollution, and has advantages of simplified operation, good feasibility, and low preparation cost. The synthesized γ-polyglutamic acid has high economic value and broad application prospect. 1. A regulation method for preparing γ-polyglutamic acid by sludge substrate fermentation , comprising the following steps:1) extraction of glutamic acid from a sludge proteinA. pretreatment: putting a sludge with a solid content of 10%-15% into a high-pressure reactor for a pretreatment to obtain a sludge slurry;B. gravity pressure filtration treatment: performing a gravity pressure filtration on the sludge slurry obtained in step A through a filter press, to obtain a dried sludge with a moisture content of 40%-60% and a first supernatant, keeping the first supernatant, and recycling the dried sludge after performing a harmless treatment on the dried sludge;2) synthesis of the γ-polyglutamic acid by a secondary metabolism{'i': Bacillus', 'Bacillus;, 'C. activation of domesticated strains: using a standard seed medium to activate and cultivate fermentation strains to obtain activated fermentation strains, and then using a fermentation medium added with fermentation raw materials to domesticate and cultivate the activated fermentation strains to obtain domesticated fermentation strains, and to amplify and propagate the domesticated ...

Generation of acyl amino acids

Engineered polypeptides useful in synthesizing acyl amino acids are provided. Also provided are methods of making acyl amino acids using engineered polypeptides. In certain embodiments, an acyl amino acid produced using compositions and/or methods of the present invention comprises cocoyl glutamate.

Xylose isomerases and their uses

This disclosure relates to novel xylose isomerases and their uses, particularly in fermentation processes that employ xylose-containing media.

NOVEL COMPOUNDS

Disclosed are compounds of the general formula I and II (as further defined herein) are useful in the production of inhibitors of sphingolipid synthesis the production of sphingolipids. Suitable sphingolipids, include, but not limited to, sphingosine and compounds incorporating sphingosine or that may use sphingosine as an intermediate or a starting material in their synthesis (including, but not limited to, sphingosine-1-P, ceramide, gangliosides and sphigomyelin). In one contemplated use, compounds of the general formula I and II are useful in the production of sphingosine. In another contemplated use, compounds of the general formula I and II are useful in the production of a sphingofugin. Methods of manufacturing each of the above compounds are also provided. 227-. (canceled)29. The compound of claim 28 , wherein Ris a silyl ether claim 28 , an alkyl ether claim 28 , an alkoxymethyl ether claim 28 , a tetrahydropyranyl ether claim 28 , a methylthiomethyl ethers claim 28 , an ester or a carbonate.30. The compound of claim 28 , wherein Ris OR claim 28 , wherein Ris a substituted or unsubstituted alkyl claim 28 , a substituted or unsubstituted alkenyl group claim 28 , a substituted or unsubstituted alkynyl group claim 28 , a substituted or unsubstituted aryl claim 28 , a substituted or unsubstituted aralkyl a substituted or unsubstituted heterocycle or a substituted or unsubstituted heterocycloalkyl.31. The compound of claim 28 , wherein Ris OR claim 28 , wherein Ris a substituted or unsubstituted alkyl claim 28 , a substituted or unsubstituted alkenyl group claim 28 , or a substituted or unsubstituted alkynyl group.32. The compound of claim 28 , wherein Ris an O—CHgroup or a benzyl group.33. The compound of claim 28 , wherein Ris selected from the group consisting of: H claim 28 ,{'sub': 2', '2', 'm', '3', '3', '3', 'm', '2', 'm', '2', '2', 'm', '2', '3', '2', 'm', '2', 'm', '3', '2', 'm', '2', '2', 'm', '2', '2', '2', 'm', '2', '2, '—CH(CH)(CH)(CH), —CH(CH)(CH2) ...

METHOD FOR PRODUCING ACRYLAMIDE AQUEOUS SOLUTION

There is provided a method for producing an aqueous acrylamide solution by producing acrylamide by reacting a composition including acrylonitrile with water, in which the composition including acrylonitrile includes 20 to 80 mg of methacrylonitrile per 1 kg of the total weight of the composition including acrylonitrile. According to the present invention, a production method allowing stable obtainment of an aqueous acrylamide solution can be provided as polymerization of the acrylamide can be suppressed without causing a decreas+455e in quality. 1. A method for producing an aqueous acrylamide solution , the method comprising reacting a composition comprising acrylonitrile with water to produce acrylamide ,wherein the composition comprises 20 to 80 mg of methacrylonitrile per 1 kg of a total weight of the composition.2. The method according to claim 1 , wherein the reacting of the composition with water is performed in the presence of a biocatalyst.3. The method for according to claim 1 , wherein the composition further comprises 2 to 20 mg of acetonitrile per 1 kg of the total weight of the composition.4. The method for according to claim 1 , wherein a concentration of the acrylamide in the aqueous acrylamide solution is from 30 to 60% by mass relative to a total mass of the aqueous acrylamide solution. The present invention relates to a method for producing an aqueous acrylamide solution.The present application claims priority to Japanese Patent Application No. 2011-112429, which has been filed in Japan on May 19, 2011, which is hereby incorporated by reference in its entirety.Acrylamide has many applications, such as flocculating agents, petroleum recovering agents, paper strength enhancers in the paper producing industry, and thickeners for papermaking, and is a useful substance as a raw material for polymers.Among industrial processes for acrylamide production, used long time ago is a sulfuric acid hydrolysis process which consists of the step of heating ...

ENGINEERED IMINE REDUCTASES AND METHODS FOR THE REDUCTIVE AMINATION OF KETONE AND AMINE COMPOUNDS

The present disclosure provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds. 1. An engineered polynucleotide encoding an engineered polypeptide having imine reductase activity , wherein said engineered polypeptide has at least 90% sequence identity to SEQ ID NO:2 , wherein the amino acid residue at the position corresponding to position 198 of SEQ ID NO:2 has been replaced with an aliphatic , non-polar , polar , or acidic residue.2. The engineered polynucleotide of claim 1 , wherein said encoded engineered polypeptide has at least 90% sequence identity to SEQ ID NO:2 claim 1 , comprising a substitution of alanine claim 1 , histidine claim 1 , proline claim 1 , serine claim 1 , or glutamic acid at position 198. The present application is a Continuation of U.S. patent application Ser. No. 15/048,887, filed Feb. 19, 2016, which is a Continuation of U.S. patent application Ser. No. 14/887,943, filed Oct. 20, 2015, now U.S. Pat. No. 9,296,993, which is a Divisional of U.S. patent application Ser. No. 13/890,944, filed May 9, 2013, now U.S. Pat. No. 9,193,957, which claims benefit under 35 U.S.C. §119(e) of U.S. Pat. Appln. Ser. No. 61/646,100, filed May 11, 2012, the contents of all of which are incorporated herein by reference.The disclosure relates to engineered polypeptides having imine reductase activity useful for the conversion of various ketone and amine substrates to secondary and tertiary amine products.The official copy of the Sequence Listing is submitted concurrently with the specification as an ASCII formatted text file via EFS-Web, with a file name of “CX2-120U52_ST25.txt”, a creation date of May 8, 2013, and a size of 1,729,414 bytes. The Sequence Listing filed ...

Ketol-Acid Reductoisomerase Enzymes and Methods of Use

Provided herein are polypeptides having ketol-acid reductoisomerase activity as well as microbial host cells comprising such polypeptides. Polypeptides provided herein may be used in biosynthetic pathways, including, but not limited to, isobutanol biosynthetic pathways. 132-. (canceled)33. A polypeptide having ketol-acid reductoisomerase activity , wherein the polypeptide comprisesa. the amino acid sequence of SEQ ID NO: 42;b. at least 99% identity to SEQ ID NO: 42;c. at least 99% identity to SEQ ID NO: 42 and at least one of the following substitutions: M3011, Y239H, Y113F, S322A, A71K, N114G, A329R, A69T, N114S, G72W, A295V, E264K, R280D, A329E, S157T, M238I, Q272T, K335M, R280H, and a combination thereof; ord. an active fragment of any one of (a) to (c).34. The polypeptide of further comprising a substitution at at least one of position L33 claim 33 , P47 claim 33 , F50 claim 33 , F61 claim 33 , 180 claim 33 , and V156.35. The polypeptide of further comprising at least one of the following substitutions: I13L claim 33 , P47Y claim 33 , F50A claim 33 , V53A claim 33 , S86A claim 33 , A268E claim 33 , V76I claim 33 , L88V claim 33 , and a combination thereof.36. A recombinant microbial host cell comprising the polypeptide of .37. The recombinant microbial host cell of claim 36 , wherein the recombinant microbial host cell further comprises reduced or eliminated acetolactate reductase activity.38. The recombinant microbial host cell of claim 37 , wherein the recombinant microbial host cell further comprises at least one deletion claim 37 , mutation claim 37 , and/or substitution in fra2.39. The recombinant microbial host cell of further comprising the substrate to product conversions:a. pyruvate to acetolactateb. 2,3-dihydroxyisovalerate to α-ketoisovaleratec. α-ketoisovalerate to isobutyraldehyde; andd. isobutyraldehyde to isobutanol.40. The recombinant microbial host cell of claim 36 , wherein the recombinant microbial host cell is a yeast host cell. ...

Methods for Making L-Glufosinate

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2-amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased. 1. A composition comprising D-glufosinate , PPO , and L-glufosinate , wherein L-glufosinate is present in the composition at an amount of 80% by weight or greater based on the total amount of D-glufosinate , PPO , and L-glufosinate.2. The composition of claim 1 , wherein the amount of L-glufosinate is 90% by weight or greater based on the total amount of D-glufosinate claim 1 , PPO claim 1 , and L-glufosinate.3. The composition of claim 1 , wherein the amount of L-glufosinate is 95% by weight or greater based on the total amount of D-glufosinate claim 1 , PPO claim 1 , and L-glufosinate.4. The composition of claim 1 , wherein the composition is a dried powder.5. A method for selectively controlling weeds in an area comprising applying an effective amount of the composition of .6. A method for selectively controlling weeds in an area comprising applying an effective amount of the composition of .7. A method for selectively controlling weeds in an area comprising applying an effective amount of the composition of . This application is a division of U.S. application Ser. No. 15/445,254, filed Feb. 28, 2017, which claims priority to U.S. Provisional Application No. 62/302,421, filed Mar. 2, 2016; U.S. Provisional Application No. 62/336,989, filed May 16, 2016; and U.S. Provisional Application No. 62/413,240, filed Oct. 26, 2016. These applications are incorporated herein by reference in their ...

ENGINEERED IMINE REDUCTASES AND METHODS FOR THE REDUCTIVE AMINATION OF KETONE AND AMINE COMPOUNDS

The present disclosure provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds. 1. An engineered polypeptide having imine reductase activity , wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2 , and comprises a substitution at a position corresponding to position 156 of SEQ ID NO:2 , wherein the amino acid at position 156 has been replaced with a non-polar , aliphatic , or polar residue.2. The engineered polypeptide having imine reductase activity of claim 1 , wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2 claim 1 , wherein the amino acid at the position corresponding to position 156 of SEQ ID NO:2 has been replaced with threonine.8. An engineered polynucleotide encoding the engineered polypeptide of .9. A vector comprising the engineered polynucleotide of .10. The vector of claim 9 , further comprising at least one control sequence.11. A host cell comprising the vector of .12. A host cell comprising the vector of . The present application is a continuation of co-pending U.S. patent application Ser. No. 16/391,036, filed on Apr. 22, 2019, which is a continuation of U.S. patent application Ser. No. 16/195,480, filed on Nov. 19, 2018, now U.S. Pat. No. 10,308,966, which is a divisional of U.S. patent application Ser. No. 16/054,843, filed on August 3, 2018, now U.S. Pat. No. 10,160,983, which is a Continuation of co-pending U.S. patent application Ser. No. 15/899,834, filed on Feb. 20, 2018, now U.S. Pat. No. 10,066,250, which is a Continuation of U.S. patent application Ser. No. 15/792,446, filed Oct. 24, 2017, now U.S. Pat. No. 9,932,613, which is a Continuation of U.S. patent application Ser. No. 15/710,462, filed Sep. 20, ...

NITRILE HYDRATASE

Improving wild nitrile hydratase enables the provision of a protein which has nitrile hydratase activity and which has further improved heat resistance, amide compound resistance and high temperature accumulation properties. Use protein (A) or (B), (A) being a protein characterised by having nitrile hydratase activity and by including an amino acid sequence in which a specific amino acid residue in an amino acid sequence in wild nitrile hydratase has been substituted by another amino acid residue, and (B) being a protein characterised by having nitrile hydratase activity and by including an amino acid sequence in which one or several amino acid residues in the amino acid sequence of protein (A), other than the abovementioned specific amino acid residue, is deleted, substituted and/or added. 1. A protein (A) or (B):(A) a protein, comprising nitrile hydratase activity,wherein in the amino-acid sequence of a wild-type nitrile hydratase, amino-acid residues at positions 167, 219, 57, 114, 107, and 17 counted downstream from an N-terminal amino-acid residue in an amino-acid sequence of a β subunit are substituted with other amino-acid residues, andan amino-acid residue at position 218, 190, 168, 144, 133, 112, 105, 95, or 15 counted downstream from an N-terminal amino-acid residue in an amino-acid sequence of a β subunit, an amino acid residue at position 67 or 17 counted downstream from a furthermost-downstream side C residue of an amino-acid sequence C(S/T)LCSC that forms a binding site with a prosthetic molecule in an amino-acid sequence of an α subunit, or any combination thereof is substituted with another amino-acid residue; or(B) a protein, comprising nitrile hydratase activity,wherein one or a plurality of amino-acid residues are deleted, substituted, added, or any combination thereof in an amino-acid sequence of protein (A) in addition to amino-acid residue substitutions of protein (A).2. An isolated DNA encoding the protein according to .3. An isolated DNA ...

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO:136 , and one or more amino acid substitutions at one or more positions in SEQ ID NO: 136 , selected from the group consisting of 37 , 277 , 278 , 280 , 281 , 326 , 432 , 433 , 435 , and 490.3. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 136 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO:136.4. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring further is capable of converting the acid substrate of compound (1b) to the R-enantiomer compound (2b) in at least 50% enantiomeric excess.5. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 136 claim 1 , wherein said polypeptide further comprises one or more substitutions selected from the group consisting of a glycine at position 143 claim 1 , glycine at ...

DILUTE CHEMICAL REACTION PROCESS

Disclosed is a process for carrying out a cyclisation reaction, a polymerization reaction, an enzymatic reaction showing substrate inhibition, an enzymatic reaction showing product inhibition, a reaction showing precipitation of the substrate or of a reactant, the process comprising the steps of 261062. The process according to claim 1 , wherein the first filtration membrane () has a rejection of at least one compound selected from the reaction product claim 1 , catalyst and one or more of the reactants which are caused to react with the substrate of 60-95% claim 1 , and wherein the at least one compound is returned from the retentate side () of the first filtration membrane () to the reactor ().36. The process according claim 1 , wherein the first membrane () has a substrate rejection of at least 95%.4. (canceled)55277222172. The process according to wherein the diluting substrate feed system () for supplying substrate to the reactor () comprises a second filtration membrane () which is permeable to the solvent (S) claim 1 , wherein the permeability of the second filtration membrane () for the substrate (X) is selected such that the permeate (P) of the second membrane has a desired concentration of the substrate (X) in the solvent (S) claim 1 , wherein permeate (P) with the desired concentration of the substrate (X) in the solvent (S) is supplied from the permeate side () of the second filtration membrane () to the reactor ().67207887. The process according to claim 5 , wherein the second filtration membrane () comprises a retentate side () claim 5 , and substrate (X) which is rejected by the second filtration membrane () is supplied to a substrate feed tank () and is further mixed with solvent claim 5 , and wherein a mixture containing solvent and substrate is supplied from the substrate feed tank () to the second filtration membrane ().711687. The process according to claim 5 , comprising the returning of solvent (S) from the permeate side () of the first ...

PREPARATION OF 6-AMINOCAPROIC ACID FROM 5-FORMYL VALERIC ACID

The invention relates to a method for preparing 6-aminocaproic acid (hereinafter also referred to as ‘6-ACA’) using a biocatalyst. The invention further relates to a method for preparing ε-caprolactam (hereafter referred to as ‘caprolactam’) by cyclising such 6-ACA. The invention further relates to a host cell, a micro-organism, or a polynucleotide which may be used in the preparation of 6-ACA or caprolactam.

METHODS FOR MAKING L-GLUFOSINATE

Methods for the production of L-glufosinate (also known as phosphinothricin or (S)-2 -amino-4-(hydroxy(methyl)phosphonoyl)butanoic acid) are provided. The methods comprise a two-step process. The first step involves the oxidative deamination of D-glufosinate to PPO (2-oxo-4-(hydroxy(methyl)phosphinoyl)butyric acid). The second step involves the specific amination of PPO to L-glufosinate, using an amine group from one or more amine donors. By combining these two reactions, the proportion of L-glufosinate in a mixture of L-glufosinate and D-glufosinate can be substantially increased.

ENGINEERED IMINE REDUCTASES AND METHODS FOR THE REDUCTIVE AMINATION OF KETONE AND AMINE COMPOUNDS

The present disclosure provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds. 1. An engineered polypeptide having imine reductase activity , wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2 , and comprises a substitution at a position corresponding to position 111 of SEQ ID NO:2 , wherein the amino acid at position 111 has been replaced with a non-polar , polar , or basic residue.2. The engineered polypeptide having imine reductase activity of claim 1 , wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2 claim 1 , wherein the amino acid at the position corresponding to position 111 of SEQ ID NO:2 has been replaced with methionine claim 1 , arginine claim 1 , glutamine claim 1 , or serine.8. An engineered polynucleotide encoding the engineered polypeptide of .9. A vector comprising the engineered polynucleotide of .10. The vector of claim 9 , further comprising at least one control sequence.11. A host cell comprising the vector of .12. A host cell comprising the vector of . The present application is a continuation of co-pending U.S. patent application Ser. No. 17/002,671, filed on Aug. 25, 2020, which is a continuation of U.S. patent application Ser. No. 16/655,547, filed on Oct. 17, 2019, now U.S. Pat. No. 10,787,689, which is a continuation of U.S. patent application Ser. No. 16/391,036, filed on Apr. 22, 2019, now U.S. Pat. No. 10,494,656, which is a continuation of U.S. patent application Ser. No. 16/195,480, filed on Nov. 19, 2018, now U.S. Pat. No. 10,308,966, which is a divisional of U.S. patent application Ser. No. 16/054,843, filed on Aug. 3, 2018, now U.S. Pat. No. 10,160,983, which is a Continuation of co-pending U ...

SINGLE STEP BIOCATALYTIC AMIDATION

The present invention relates to a process for biocatalytic amidation of non-protected amino acids comprising the step of contacting a non-protected amino acid with an ammonia source in the presence of an organic solvent and an enzyme. 1. A process for biocatalytic amidation of non-protected amino acids comprising the step of contacting a non-protected amino acid with an ammonia source in the presence of an organic solvent and an enzyme.2. The process according to claim 1 , wherein the enzyme is a lipase.3Candida antarctica. The process according to claim 2 , wherein the lipase is lipase B (CalB) or a variant thereof.4. The process according to claim 3 , wherein the CalB variant comprises the amino acid substitutions T57A/A89T/G226R/R168K with reference to the numbering of SEQ ID NO:1.5. The process according to claim 1 , wherein the enzyme is immobilized.6. The process according to claim 1 , wherein the ammonia source is ammonia (NH) claim 1 , ammonium carbamate claim 1 , ammonium carbonate claim 1 , benzylamine claim 1 , ethanolamine claim 1 , hexylamine claim 1 , 2-phenylethylamine claim 1 , or 3-phenylpropylamine claim 1 , or any mixtures thereof.7. The process according to claim 1 , wherein the organic solvent is selected from the group consisting of dioxane claim 1 , 2-methyl-2-butanol claim 1 , and t-butanol claim 1 , or from ionic liquids.8. The process according to claim 1 , wherein the unprotected amino acid is L-proline.9. The process according to claim 1 , wherein the process is carried out at a temperature in the range of 60 to 80° C.10. The process according to claim 1 , wherein ammonia is comprised in the organic solvent.11. The process according to claim 1 , wherein the water content in the reaction media is below 1.0 v/v % claim 1 , or below 0.5 v/v %.12. The process according to claim 1 , wherein the process is performed in batch or continuous mode.13. The process according to claim 12 , wherein the unprotected amino acid in the batch mode is ...

PROCESSES AND RECOMBINANT MICROORGANISMS FOR THE PRODUCTION OF CADAVERINE

Recombinant microorganisms comprising DNA molecules in a deregulated form which improve the production of cadaverine or N-acetylcadaverine, as well as recombinant DNA molecules and polypeptides used to produce the microorganisms are provided. Said microorganisms comprise an intracellular lysine decarboxylase activity and a deregulated cadaverine export activity, or comprise a decreased cadaverine export activity and an enhanced N-acetylcadaverine forming activity. Processes for the production of cadaverine N-acetylcadaverine using the recombinant microorganisms are also provided. 1. A microorganism comprising a deregulated cadaverine exporter activity.2. A microorganism as claimed in claim 1 , wherein the deregulated cadaverine exporter activity is at least partially due to deregulation of one or more cadaverine exporter polypeptides comprising an amino acid sequence being at least 80% identical to SEQ ID NO: 13. A microorganism as claimed in claim 1 , wherein the cadaverine exporter activity is enhanced.4. A microorganism as claimed in claim 1 , wherein the lysine decarboxylase activity is enhanced5. A microorganism as claimed in claim 3 , wherein the lysine decarboxylase activity is due to expression of one or more lysine decarboxylase polypeptides comprising an amino acid sequence being at least 80% identical to SEQ ID NO: 3 or 4.6. A microorganism as claimed in claim 3 , having at least one deregulated gene selected from the group consisting of the genes of aspartokinase claim 3 , aspartatesemialdehyde dehydrogenase claim 3 , dihydrodipicolinate synthase claim 3 , dihydrodipicolinate reductase claim 3 , tetrahydrodipicolinate succinylase claim 3 , succinyl-amino-ketopimelate transaminase claim 3 , succinyl-diamino-pimelate desuccinylase claim 3 , diaminopimelate epimerase claim 3 , diaminopimelate dehydrogenase claim 3 , arginyl-tRNA synthetase claim 3 , diaminopimelate decarboxylase claim 3 , pyruvate carboxylase claim 3 , phosphoenolpyruvate carboxylase claim ...

METHOD FOR PRODUCING ACRYLAMIDE

The present invention relates to a method for producing acrylamide comprising: supplying a raw material water to a reactor, supplying acrylonitrile to the reactor, and hydrating acrylonitrile using a biocatalyst, wherein a temperature of the raw material water in the supplying step of the raw material water to the reactor is equal to or more than freezing point of the raw material water and lower than the reaction temperature by 5° C. or more. The present invention can be provided to the method of producing acrylamide which can be removed the reaction heat produced at the hydration reaction of acrylonitrile effectively with a low cost. 1. A method for producing acrylamide , comprising:supplying a raw material water to a reactor,supplying acrylonitrile to the reactor, andhydrating acrylonitrile using with a biocatalyst,wherein a temperature of the raw material water in the supplying the raw material water is equal to or more than a freezing point of the raw material water and lower than a reaction temperature by 5° C. or more.2. The method according to claim 1 ,wherein a temperature of acrylonitrile in the supplying acrylonitrile to the reactor is equal to or more than the freezing point of the raw material water and lower than the reaction temperature by 5° C. or more.3. The method according to claim 1 ,wherein the hydrating acrylonitrile with the biocatalyst is performed by a semi batch reaction or a continuous reaction.4. The method according to claim 3 , wherein the hydrating acrylonitrile with the biocatalyst is performed by a semi batch reaction.5. The method according to claim 3 , wherein the hydrating acrylonitrile with the biocatalyst is performed by a continuous reaction. The present invention relates to a method for producing acrylamide from acrylonitrile using a biocatalyst. The present invention claims priority on the basis of Japanese Patent Application No. 2011-121849 filed in Japan on May 31, 2011, the contents of which are incorporated herein by ...

GENERATION OF ACYL AMINO ACIDS

In certain embodiments, the present invention comprises compositions and methods useful in the generation of acyl amino acids. In certain embodiments, the present invention provides an engineered polypeptide comprising a peptide synthetase domain; in some such embodiments, the engineered polypeptide comprises only a single peptide synthetase domain. In some embodiments, the present invention provides an engineered peptide synthetase that is substantially free of a thioesterase domain, and/or a reductase domain. In certain embodiments, the present invention provides an acyl amino acid composition comprising a plurality of different forms of an acyl amino acid. In some such compositions, substantially all of the acyl amino acids within the composition contain the same amino acid moiety and differ with respect to acyl moiety. We also described populations where the fatty acid si for example 95% one length (C14, myristic). 1. A method of making an acyl amino acid composition by contacting an engineered peptide synthetase with an amino acid substrate and an acyl entity substrate for the engineered peptide synthetase , under conditions and for a time sufficient for an acyl amino acid composition to be made.2. The method of claim 1 , wherein the engineered peptide sythetase includes an adenylation (A) domain claim 1 , a thiolation (T) domain claim 1 , and a condensation (C) domain.3. The method of claim 1 , wherein the engineered peptide synthetase lacks thioesterase domain claim 1 , and/or a reductase domain.4. The method of claim 1 , wherein the engineered peptide synthetase contains only a single peptide synthetase domain.5. The method of claim 1 , wherein the engineered peptide synthetase is or comprises a peptide synthetase domain found in as a first domain in a peptide synthetase that synthesizes a lipopeptide.6. The method of claim 1 , wherein acyl amino acid composition includes claim 1 , as a prominent component claim 1 , an acyl amino acid whose amino acid moiety ...

GENERATION OF ACYL AMINO ACIDS

Engineered polypeptides useful in synthesizing acyl amino acids are provided. Also provided are methods of making acyl amino acids using engineered polypeptides. In certain embodiments, an acyl amino acid produced using compositions and/or methods of the present invention comprises cocoyl glutamate. 1. A method of producing an acyl amino acid , comprising steps of:providing an engineered polypeptide comprising a fatty acid linkage domain, a peptide synthetase domain, and a thioesterase domain;which fatty acid linkage domain, peptide synthetase domain and thioesterase domain are covalently linked;providing fatty acid recognized by the fatty acid linkage domain;providing an amino acid recognized by the peptide synthetase domain;incubating the engineered polypeptide, fatty acid, and amino acid under conditions and for a time sufficient for the fatty acid linkage domain to link the fatty acid to the amino acid to generate an acyl amino acid;incubating the engineered polypeptide and the acyl amino acid under conditions and for a time sufficient for the thioesterase domain to catalyze release of the acyl amino acid from the engineered polypeptide.2. A method of producing an acyl amino acid , comprising steps of:providing an engineered polypeptide comprising a fatty acid linkage domain, a peptide synthetase domain, and a reductase domain;which fatty acid linkage domain, peptide synthetase domain and reductase domain are covalently linked;providing a fatty acid recognized by the fatty acid linkage domain;providing an amino acid recognized by the peptide synthetase domain;incubating the engineered polypeptide, fatty acid, and amino acid under conditions and for a time sufficient for the fatty acid linkage domain to link the fatty acid to the amino acid to generate an acyl amino acid;incubating the engineered polypeptide and the acyl amino acid under conditions and for a time sufficient for the reductase domain to catalyze release of the acyl amino acid from the engineered ...

NITRILE HYDRATASE

Provided is an improved nitrile hydratase with improved catalytic activity. Also provided are DNA for coding the improved nitrile hydratase, a recombinant vector that contains the DNA, a transformant that contains the recombinant vector, nitrile hydratase acquired from a culture of the transformant, and a method for producing the nitrile hydratase. Also provided is a method for producing an amide compound that uses the culture or a processed product of the culture. The improved nitrile hydratase contains an amino acid sequence represented by SEQ ID NO: 50 (GXXXXDXXR) in a beta subunit, and is characterized in that Xis an amino acid selected from a group comprising cysteine, aspartic acid, glutamic acid, histidine, isoleucine, lysine, methionine, asparagine, proline, glutamine, serine and threonine. 1. (canceled)2RhodococcusNocardia. A modified bacterial or bacterial nitrile hydratase , comprising in the β subunit , an amino-acid sequence as shown in SEQ ID NO: 81:{'sub': 1', '2', '3', '4', '5', '6', '7', '8', '9', '10', '11', '12', '13', '14', '15', '16', '17', '18, 'claim-text': wherein', 'W is tryptophan,', 'E is glutamic acid,', 'D is aspartic acid,', {'sub': 1', '6', '8', '13', '15', '18, 'X˜X, X˜Xand X˜Xeach independently indicate any amino-acid residue,'}, {'sub': '7', 'Xis an amino acid selected from the group consisting of alanine, valine, aspartic acid, threonine, phenylalanine, isoleucine and methionine, and'}, {'sub': '14', 'Xis glycine.'}], 'WEXXXXXXXXXXXXXXXXXXD (SEQ ID NO: 81),'}3RhodococcusNocardia. The modified bacterial or bacterial nitrile hydratase according to claim 2 , wherein Xis G (glycine) claim 2 , Xis R (arginine) claim 2 , Xis T (threonine) claim 2 , Xis L (leucine) claim 2 , Xis S (serine) claim 2 , Xis I (isoleucine) claim 2 , Xis T (threonine) claim 2 , Xis W (tryptophan) claim 2 , Xis M (methionine) claim 2 , Xis H (histidine) claim 2 , Xis L (leucine) claim 2 , Xis K (lysine) claim 2 , and Xis G (glycine) in SEQ ID NO: 81. ...

ENGINEERED IMINE REDUCTASES AND METHODS FOR THE REDUCTIVE AMINATION OF KETONE AND AMINE COMPOUNDS

The present disclosure provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds. 1. An engineered polypeptide having imine reductase activity , wherein said polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO:2 , and comprises a substitution at a position corresponding to position 280 of SEQ ID NO:2 , wherein the amino acid at position 280 has been replaced with an aliphatic or non-polar residue.2. The engineered polypeptide having imine reductase activity of claim 1 , wherein said polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO:2 claim 1 , wherein the amino acid at the position corresponding to position 280 of SEQ ID NO:2 has been replaced with leucine.9. The process of claim 8 , in which Rand Rare linked to form a 3-membered to 10-membered ring.10. The process of claim 8 , in which the substrate compound of formula (II) is selected from methylamine claim 8 , dimethylamine claim 8 , isopropylamine claim 8 , butylamine claim 8 , isobutylaminel claim 8 , L-norvaline claim 8 , aniline claim 8 , (S)-2-aminopent-4-enoic acid claim 8 , pyrrolidine claim 8 , and hydroxypyrrolidine.11. The process of claim 8 , in which at least one of Rand Rof the compound of formula (I) is linked to at least one of Rand Rof the amine compound of formula (II) claim 8 , whereby the process for preparing the amine compound of formula (III) comprises an intramolecular reaction.12. The process of claim 8 , in which the suitable reaction conditions comprise:(a) substrate loading at about 10 g/L to 100 g/L;(b) about 0.1 g/L to about 50 g/L of the engineered polypeptide;(c) about 0.05 g/L (0.001 M) to about 2.5 g/L (0.050 M) of NAD(P)H;(d) a pH of ...

Isolated Codon Sequence

The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular/modified/synthetic gene(s) of select enzymes are processed and inserted into an expression system (vector, cosmid, BAC, YAC, phage, etc.) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product. 1. A method for producing enzymes and products , wherein the method comprises:inserting a gene into an expression system and thereby yielding a modified host;optimizing the modified host and thereby yielding the enzymes and the products;purifying the enzymes and products; andwherein the gene contains a codon-optimized nucleic acid sequence of SEQ ID NO: 34 or a nucleic acid sequence that is 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 34; or an codon-optimized nucleic acid sequence of nucleic acids 5052 to 6671 of SEQ ID NO: 34 or a nucleotide sequence that is 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more homologous to the nucleic acids 5052 to 6671 of SEQ ID NO: 34.2Cannabis. The method of claim 1 , wherein the products are found in a plant.3. The method of claim 1 , wherein the products comprise at least one of: cannabinoids claim 1 , terpenoids claim 1 , stilbenoids claim 1 , flavonoids claim 1 , phenolic amides claim 1 ...

Isolated Codon Sequence

The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular/modified/synthetic gene(s) of select enzymes are processed and inserted into an expression system (vector, cosmid, BAC, YAC, phage, etc.) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product. 1. A host organism , wherein the host organism comprises:the host organism that is in a modified form in comparison to a natural state of the host organism via gene insertion, and thereby contains an increased yield of product in comparison to a yield of product in the natural state of the host organism;a plurality of pathways in the host organism, wherein a first pathway of the plurality of pathways generates the product;a plurality of substrates for deriving the product; anda codon-optimized nucleic acid sequence of SEQ ID NO: 34 or a nucleic acid sequence that is 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 34;2. The host organism of claim 2 , wherein the host organism is a bacteria claim 2 , plant claim 2 , alga claim 2 , or fungus.3S. cerevisiae, E. coli, P. pastorisis, N. crassa, S. pombe, R. palmatum, C. Tonga, O. sativa, A. arborescens, A. terrus, C. sativa, S. griseus, S. erythera, S. coelicolor, S. toxytruini, S. cellulosum, P. ...

BIOCATALYSTS FOR THE PREPARATION OF HYDROXY SUBSTITUTED CARBAMATES

The present disclosure relates to engineered ketoreductase polypeptides for the preparation of hydroxyl substituted carbamate compounds, and polynucleotides, vectors, host cells, and methods of making and using the ketoreductase polypeptides. 2. The engineered polynucleotide encoding an engineered ketoreductase polypeptide of claim 1 , wherein the amino acid sequence of said ketoreductase polypeptide comprises the substitution X206F/L and one or more residue differences as compared to SEQ ID NO:4 selected from: X7S; X17M; X17Q; X17R; X23V; X27L; X29G; X40R; X60I; X64V; X71P; X87L; X94A; X94P; X94S; X95M; X105G; X113I; X122A; X127R; X131S; X144V; X145L; X147I; X147L; X147Q; X150Y; X152G; X153G; X157C; X173L; X196M; X198S; X208R; X216R; X221S; X243S; X245I; X249F; X249G; and X249Y.3. The engineered polynucleotide encoding an engineered ketoreductase polypeptide of claim 1 , wherein the amino acid sequence of said ketoreductase polypeptide comprises X206F/L claim 1 , and at least one or more residue differences as compared to SEQ ID NO:4 selected from: X17Q/R/M; X40R; X64V; X94P; X96L/Y; X144V; X147Q/I/L; X157C; X195A/G; X196M; and X199H.4. The engineered polynucleotide encoding an engineered ketoreductase polypeptide of claim 1 , wherein the ketoreductase polypeptide is capable of converting the substrate compound (2) to the product compound (1) with at least 10 fold the activity of the reference polypeptide of SEQ ID NO:4 claim 1 , wherein the amino acid sequence comprises the substitution X206F/L claim 1 , and one or more residue differences as compared to SEQ ID NO:4 selected from: X40R; X60I; X71P; X94P; X94A; X95M; X96L; X96Y; X127R; X144V; X145I; X150Y; X152G; X153G; X157C; X195A; X195G; X196M; X198S; X199H; X216R claim 1 , X245I claim 1 , X245F; X249Y; and X249F.5. The engineered polynucleotide encoding an engineered ketoreductase polypeptide of claim 1 , wherein the ketoreductase polypeptide has increased thermal stability as compared to the reference ...

GENERATION OF ACYL AMINO ACIDS

Engineered polypeptides useful in synthesizing acyl amino acids are provided. Also provided are methods of making acyl amino acids using engineered polypeptides. 1. A method of making an acyl amino acid composition by contacting an engineered peptide synthetase with an amino acid substrate and an acyl entity substrate for the engineered peptide synthetase , under conditions and for a time sufficient for an acyl amino acid composition to be made.2. The method of claim 1 , wherein the engineered peptide synthetase includes an adenylation (A) domain claim 1 , a thiolation (T) domain claim 1 , and a condensation (C) domain.3. The method of claim 1 , wherein the engineered peptide synthetase lacks thioesterase domain claim 1 , and/or a reductase domain.4. The method of claim 1 , wherein the engineered peptide synthetase contains only a single peptide synthetase domain.5. The method of claim 1 , wherein the engineered peptide synthetase is or comprises a peptide synthetase domain found in as a first domain in a peptide synthetase that synthesizes a lipopeptide.6. The method of claim 1 , wherein acyl amino acid composition includes claim 1 , as a prominent component claim 1 , an acyl amino acid whose amino acid moiety is from an amino acid selected from the group consisting of glycine or glutamate and whose acyl moiety is from a fatty acid selected from the group consisting of myristic acid and or lauric acid.7. The method of claim 1 , wherein the step of contacting comprises providing a cell engineered to express at least one engineered peptide synthetase.8. A cell engineered to express at least one engineered peptide synthetase that synthesizes an acyl amino acid.9. An acyl amino acid composition produced by an engineered peptide synthetase.10. The composition of claim 9 , wherein substantially all of the acyl amino acids in the composition contain the same amino acid component.11. The composition of claim 9 , wherein acyl amino acids in the composition comprise ...

MICROBIAL PRODUCTION OF ALKANOLAMIDES AND AMIDOAMINES AND USES THEREOF

The disclosure relates to a recombinant microorganism engineered to express an enzyme which catalyzes the conversion of a primary amine and an acyl thioester to a fatty amide. The disclosure further encompasses a method of producing a fatty amide by culturing the recombinant microorganism in the presence of a carbon source. 1. A recombinant microorganism comprising a nucleic acid sequence encoding a polypeptide that catalyzes a conversion of a primary amine and an acyl thioester to a fatty amide , wherein the microorganism is cultured in the presence of a carbon source.2. The recombinant microorganism of claim 1 , wherein the polypeptide is a palmitoylputrescine synthase (PPS) polypeptide.3. The recombinant microorganism of claim 2 , wherein the PPS polypeptide comprises an amino acid sequence that has at least about 70% claim 2 , at least about 75% claim 2 , at least about 80% claim 2 , at least about 85% claim 2 , at least about 90% claim 2 , at least about 91% claim 2 , at least about 92% claim 2 , at least about 93% claim 2 , at least about 94% claim 2 , at least about 95% claim 2 , at least about 96% claim 2 , at least about 97% claim 2 , at least about 98% claim 2 , or at least about 99% sequence identity to an amino acid sequence of SEQ ID NO: 1.4. The recombinant microorganism of claim 2 , wherein the PPS polypeptide has an amino acid sequence comprising SEQ ID NO: 1.5. The recombinant microorganism of claim 2 , wherein the PPS polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 2.6. The recombinant microorganism of claim 1 , wherein the polypeptide is a N-(4-amino-2-hydroxylbutyl)tetradecanamide synthase (AhtS) polypeptide.7. The recombinant microorganism of claim 6 , wherein the AhtS polypeptide comprises an amino acid sequence that has at least about 70% claim 6 , at least about 75% claim 6 , at least about 80% claim 6 , at least about 85% claim 6 , at least about 90% claim 6 , at least about 91% claim 6 , at least about 92% claim 6 , ...

Method for Producing 3-Acetylamino-4-Hydroxybenzoic Acid

The present invention provides a method for conveniently and efficiently producing a 3-acetylamino-4-hydroxybenzoic acid-type compound that is a stable compound by a process using a microorganism. Specifically the present invention provides a microorganism having an ability to produce 3-amino-4-hydroxybenzoic acid, that is modified so as to increase an activity to form 3-amino-4-hydroxybenzoic acid from dihydroxyacetone phosphate and aspartate semialdehyde, wherein the microorganism is modified so as to increase an N-hydroxyarylamine O-acetyltransferase (NhoA) activity, as well as a method for producing the 3-acetylamino-4-hydroxybenzoic acid-type compound using such a microorganism. 1. A microorganism that is able to produce 3-amino-4-hydroxybenzoic acid , wherein the microorganism is modified so as to increase formation of 3-amino-4-hydroxybenzoic acid from dihydroxyacetone phosphate and aspartate semialdehyde , wherein the microorganism is modified so as to increase an N-hydroxyarylamine O-acetyltransferase (NhoA) activity.2. The microorganism according to claim 1 , wherein the N-hydroxyarylamine O-acetyltransferase activity is increased by transformation with a recombinant vector comprising a DNA encoding the NhoA.3. The microorganism according to claim 2 , wherein the NhoA is a protein selected from the group consisting of:(I) a protein comprising the amino acid sequence of SEQ ID NO:2;(II) a protein comprising an amino acid sequence having one or several amino acid substitutions, deletions, insertions or additions in the amino acid sequence of SEQ ID NO:2 and having an N-hydroxyarylamine O-acetyltransferase activity; and(III) a protein comprising an amino acid sequence having 70% or more identity to the amino acid sequence of SEQ ID NO:2 and having an N-hydroxyarylamine O-acetyltransferase activity.4. The microorganism according to claim 2 , wherein the DNA encoding the NhoA is a DNA selected from the group consisting of:(i) a DNA comprising the nucleotide ...

ENGINEERED IMINE REDUCTASES AND METHODS FOR THE REDUCTIVE AMINATION OF KETONE AND AMINE COMPOUNDS

The present disclosure provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds. 1. An engineered polynucleotide encoding an engineered polypeptide having imine reductase activity , wherein said polypeptide has at least 90% sequence identity to the polypeptide SEQ ID NO:2 , and comprises a substitution at a position corresponding to position 293 of SEQ ID NO:2 , wherein the amino acid at position 293 has been replaced with an aromatic , non-polar , aliphatic , basic , or polar residue.2. The engineered polynucleotide encoding the engineered polypeptide having imine reductase activity of claim 1 , wherein said polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO:2 claim 1 , wherein the amino acid at the position corresponding to position 293 of SEQ ID NO:2 has been replaced with leucine.9. The process of claim 8 , in which Rand Rare linked to form a 3-membered to 10-membered ring.10. The process of claim 8 , in which the substrate compound of formula (II) is selected from methylamine claim 8 , dimethylamine claim 8 , isopropylamine claim 8 , butylamine claim 8 , isobutylaminel claim 8 , L-norvaline claim 8 , aniline claim 8 , (S)-2-aminopent-4-enoic acid claim 8 , pyrrolidine claim 8 , and hydroxypyrrolidine.11. The process of claim 8 , in which at least one of Rand Rof the compound of formula (I) is linked to at least one of Rand Rof the amine compound of formula (II) claim 8 , whereby the process for preparing the amine compound of formula (III) comprises an intramolecular reaction.12. The process of claim 8 , in which the suitable reaction conditions comprise:(a) substrate loading at about 10 g/L to 100 g/L;(b) about 0.1 g/L to about 50 g/L of the ...

Amphoteric compounds

Disclosed are a variety of amphoteric compounds containing a quaternary nitrogen group, a covalently bound counterion, and an ester or amide group. These amphoteric compounds can be advantageously prepared via a chemoenzymatic green process, and exhibit good surfactant properties.

Novel system based on a new nitrile hydratase for highly efficient catalytic hydration reaction of aliphatic dinitriles

The invention belongs to the technical field of green chemistry, and provides a novel system based on a new nitrile hydratase for highly efficient catalytic conversion of aliphatic dinitriles. The invention discloses a new application of nitrile hydratase using CCM 2595 in catalyzing aliphatic dinitrile. In particular, the enzyme can regioselectivity catalyze the formation of 5-cyanopyramides from adiponitrile with high reaction rate under mild reaction conditions, which provides a new method for the industrial production of 5-cyanopyramides. 1Rhodococcus erythropohs. A novel system based on a new nitrile hydratase for highly efficient catalytic hydration reaction of aliphatic dinitriles , wherein the concentration of the recombinant bacteria with nitrile hydratase derived from the CCM2595 is 1-3 g/L , the concentration of aliphatic dinitriles is 20-50 mM/L , and conversion system comprises phosphate buffer saline solution with pH 7-8 , temperature of 25° C. and oscillation with 200 rpm for reaction; the reaction is then quenched by adding equal volume of methanol after 5 min cultivation; then supernatant is collected after centrifugation; the supernatant is filtered for high performance liquid chromatography detection; the regioselectivity of the recombinant bacteria with nitrile hydratase towards aliphatic dinitriles is more than 90%.2. The system according to claim 1 , wherein the preparation steps of the recombinant bacteria with nitrile hydratase are as follows:{'i': Rhodococcus erythropohs', 'E. coli, '(1) plasmid construction: the gene sequence of nitrile hydratase from the strain CCM2595 contains 2596 nucleotides; plasmid pET-24a (+) is used as the expression vector, according to the characteristics of restriction sites of the plasmid, NdeI and Hind III restriction sites are selected to insert the nitrile hydratase gene which is obtained by PCR; after digestion, the corresponding DNA fragment is recovered and purified, and inserted the kanamycin KanR ...

Nitrile hydratase

Provided is an improved nitrile hydratase with improved catalytic activity. Also provided are DNA for coding the improved nitrile hydratase, a recombinant vector that contains the DNA, a transformant that contains the recombinant vector, nitrile hydratase acquired from a culture of the transformant, and a method for producing the nitrile hydratase. Also provided is a method for producing an amide compound that uses the culture or a processed product of the culture. The improved nitrile hydratase contains an amino acid sequence represented by SEQ ID NO: 50 (GX1X2X3X4DX5X6R) in a beta subunit, and is characterized in that X4 is an amino acid selected from a group comprising cysteine, aspartic acid, glutamic acid, histidine, isoleucine, lysine, methionine, asparagine, proline, glutamine, serine and threonine.

Isoprene oligomer, polyisoprene, processes for producing these materials, rubber composition, and pneumatic tire

The invention relates to an isoprene oligomer that contains a trans structural moiety and a cis structural moiety, which can be represented by the following formula (1), wherein at least 1 atom or group in the trans structural moiety is replaced by another atom or group. The invention also relates to a polyisoprene, which is biosynthesized using the isoprene oligomer and isopentenyl diphosphate. Further, this invention provides a rubber composition comprising the isoprene oligomer and/or the polyisoprene, and a pneumatic tire, including tire components (e.g., treads and sidewalls) formed from the rubber composition. wherein n represents an integer from 1 to 10; m represents an integer from 1 to 30; and Y represents a hydroxy group, a formyl group, a carboxy group, an ester group, a carbonyl group, or a group represented by the following formula (2):

NITRILE HYDRATASE

Provided is an improved nitrile hydratase with improved catalytic activity. Also provided are DNA for coding the improved nitrile hydratase, a recombinant vector that contains the DNA, a transformant that contains the recombinant vector, nitrile hydratase acquired from a culture of the transformant, and a method for producing the nitrile hydratase. Also provided is a method for producing an amide compound that uses the culture or a processed product of the culture. The improved nitrile hydratase contains an amino acid sequence represented by SEQ ID NO: 50 (GXXXXDXXR) in a beta subunit, and is characterized in that Xis an amino acid selected from a group comprising cysteine, aspartic acid, glutamic acid, histidine, isoleucine, lysine, methionine, asparagine, proline, glutamine, serine and threonine. 1. (canceled)4RhodococcusNocardia. The modified bacterial or bacterial nitrile hydratase according to claim 3 , wherein Xis M (methionine) claim 3 , Xis A (alanine) claim 3 , Xis S (serine) claim 3 , Xis L (leucine) claim 3 , Xis Y (tyrosine) claim 3 , Xis A (alanine) and Xis E (glutamic acid) in SEQ ID NO: 119.5RhodococcusNocardia. The modified bacterial or bacterial nitrile hydratase according to claim 3 , further comprising an amino-acid sequence in SEQ ID NO: 120 comprising the amino-acid sequence in SEQ ID NO: 119.7RhodococcusNocardia. The modified bacterial or bacterial nitrile hydratase according to claim 6 , further comprising an amino-acid sequence of the α subunit in SEQ ID NO: 132 claim 6 , wherein Xis an amino acid selected from the group consisting of cysteine claim 6 , phenylalanine claim 6 , histidine claim 6 , isoleucine claim 6 , lysine claim 6 , methionine claim 6 , glutamine claim 6 , arginine claim 6 , threonine and tyrosine.8RhodococcusNocardia. The modified bacterial or bacterial nitrile hydratase according to claim 6 , wherein Xis M (methionine) claim 6 , Xis A (alanine) claim 6 , Xis S (serine) claim 6 , Xis L (leucine) claim 6 , Xis Y (tyrosine) ...

PREPARATION OF 6-AMINOCAPROIC ACID FROM 5-FORMYL VALERIC ACID

The invention relates to a method for preparing 6-aminocaproic acid (hereinafter also referred to as ‘6-ACA’) using a biocatalyst. The invention further relates to a method for preparing ϵ-caprolactam (hereafter referred to as ‘caprolactam’) by cyclising such 6-ACA. The invention further relates to a host cell, a micro-organism, or a polynucleotide which may be used in the preparation of 6-ACA or caprolactam. 1. (canceled)2. Method for preparing 6-aminocaproic acid , wherein the 6-aminocaproic acid is prepared from 5-formylpentanoate , using at least one biocatalyst.325-. (canceled) This application is a divisional of U.S. patent application Ser. No. 14,105,705, filed Dec. 13, 2013, which is a continuation of U.S. patent application Ser. No. 12/921,733, filed Dec. 21, 2010, now issued U.S. Pat. No. 8,673,599, issued Mar. 18, 2014, which is a U.S. National Stage Application under 35 U.S.C. § 371 of International Patent Application No. PCT/NL2009/050117, filed Mar. 11, 2009, which claims the benefit of European Patent Application No. 08152584.2, filed Mar. 11, 2008, the entire contents of which are each incorporated herein by reference.The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on May 17, 2017, is named Sequence_Listing_12956-416-999.txt and is 249,748 bytes in size.The invention relates to a method for preparing 6-aminocaproic acid (hereinafter also referred to as ‘6-ACA’). The invention further relates to a method for preparing ϵ-caprolactam (hereafter referred to as ‘caprolactam’) from 6-ACA. The invention further relates to a host cell which may be used in the preparation of 6-ACA or caprolactam.Caprolactam is a lactam which may be used for the production of polyamide, for instance nylon-6 or nylon-6,12 (a copolymer of caprolactam and laurolactam). Various manners of preparing caprolactam from bulk chemicals are known in ...

METHOD AND APPARATUS FOR PRODUCING ACRYLAMIDE

Provided is a technique which can easily realize the retention time of the reaction mixture in the reactor suitable for the production quantity by controlling the amount of the reaction liquid in accordance with the production quantity and thus can suppress the amount of a biocatalyst used in a method for producing acrylamide from acrylonitrile by using a biocatalyst. Provided is a method for producing acrylamide from acrylonitrile through a continuous reaction using a biocatalyst in reactors by using two or more reactors connected in series, wherein one reactor A and a reactor B connected to the reactor A on an upstream side are communicated with each other below liquid faces of reaction liquids in both reactors, and the producing method comprises controlling a liquid volume of a reaction liquid in the reactor B by controlling a level of a reaction liquid in the reactor A to be between a disposed position of a communicating port with the reactor B and a full level position. 1: A method for producing acrylamide from acrylonitrile through a continuous reaction using a biocatalyst in two or more reactors connected in series , the method comprisingcontrolling a liquid volume of a reaction liquid in a reactor B by controlling a level of a reaction liquid in a reactor A to be between a disposed position of a communicating port with the reactor B and a full level position,whereinthe reactor B is connected to the reactor A on an upstream side, andthe reactor A and the reactor B are communicated with each other below liquid faces of the reaction liquids in both reactors.2: The producing method according to claim 1 , whereinthe reactor A includes a circulating line to circulate the reaction liquid and a discharge line to discharge the reaction liquid, andthe level of the reaction liquid in the reactor A is controlled by adjusting a liquid volume of the reaction liquid to be discharged and/or a liquid volume of the reaction liquid to return to the reactor A through ...

Activity of Fe-S Cluster Requiring Proteins

The present invention is related to a recombinant host cell, in particular a yeast cell, comprising a dihydroxy-acid dehydratase polypeptide. The invention is also related to a recombinant host cell having increased specific activity of the dihydroxy-acid dehydratase polypeptide as a result of increased expression of the polypeptide, modulation of the Fe—S cluster biosynthesis of the cell, or a combination thereof. The present invention also includes methods of using the host cells, as well as, methods for identifying polypeptides that increase the flux in an Fe—S cluster biosynthesis pathway in a host cell.

Xylose Isomerases and Their Uses

This disclosure relates to novel xylose isomerases and their uses, particularly in fermentation processes that employ xylose-containing media.

BACTERIAL STRAIN RHODOCOCCUS AETHERIVORANS VKM Ac-2610D PRODUCING NITRILE HYDRATASE, METHOD OF ITS CULTIVATION AND METHOD FOR PRODUCING ACRYLAMIDE

The invention relates to a bacterial strain belonging to the genus which is a producer of a nitrile hydratase. The invention also relates to a method for producing acrylamide by hydration of acrylonitrile using a biomass of the bacterial strain and to a method of culturing the bacterial strain. 1Rhodococcus aetherivorans. A bacterial strain VKM Ac-2610D.2. The bacterial strain according to claim 1 , which is a producer of a nitrile hydratase.3RhodococcusRhodococcus aetherivorans. A method for producing acrylamide by hydration of acrylonitrile using a biomass of a strain belonging to the genus and having a nitrile hydratase activity claim 1 , wherein the hydration is carried out at a working concentration of acrylonitrile which is no more than 0 claim 1 ,5% claim 1 , using a biomass of the bacterial strain VKM Ac-2610D.4Rhodococcus aetherivorans. The method for producing acrylamide according to claim 3 , wherein acrylonitrile is mixed in an aqueous suspension of a biomass of the bacterial strain VKM Ac-2610D to form a reaction solution; and hydration is carried out so that the working concentration of acrylonitrile in the reaction solution is maintained at no more than 0 claim 3 ,5%.5. The method for producing acrylamide according to claim 3 , wherein the hydration is followed by isolation of the final acrylamide product.6Rhodococcus aetherivorans. The method for producing acrylamide according to claim 3 , wherein the biomass of the bacterial strain VKM Ac-2610D is at about 100-1000 g claim 3 , or 200-1000 g claim 3 , or 200-800 g claim 3 , or 300-600 g claim 3 , or about 400-500 g claim 3 , or less than 1000 g claim 3 , or less than 500 g claim 3 , or less than 400 g on a dry weight of strain per 1 ton of the final product.7. The method for producing acrylamide according to claim 6 , wherein the final product contains acrylamide at a concentration of at least 40%; or at least 41%; or at least 42%; or at least 43%; or at least 44%; or 40-55%; or 41-55%; 42-55%; or 43 ...

ACETYLTRANSFERASE FROM WICKERHAMOMYCES CIFERRII

The invention relates to novel enzymes that provide acetylated sphingoid bases. 1. An isolated nucleic acid having a sequence selected from:a sequence with at least 90% identity to SEQ ID NO: 1 and comprising at least one substitution, addition, inversion and/or deletion relative to SEQ ID NO: 1, anda sequence which hybridizes with SEQ ID NO: 1 under stringent conditions.2. An isolated nucleic acid having a sequence selected from:a sequence with at least 90% identity to SEQ ID NO: 3 and comprising at least one substitution, addition, inversion and/or deletion relative to SEQ ID NO: 3, anda sequence which hybridizes with SEQ ID NO: 3 under stringent conditions.3. A genetically modified cell , the genetic modification comprising the introduction of one or more expression vectors resulting in increased expression at least one enzyme comprising:the polypeptide sequence of SEQ ID NO: 2or the polypeptide sequence of SEQ ID NO: 4.4. The genetically modified cell of claim 3 , wherein said genetic modification results in increased expression of the enzyme of SEQ ID NO: 2 and the enzyme of SEQ ID NO: 4.5Saccharomyces cerevisiae, Kluyveromyces lactis, Hansenula polymorpha, Pichia pastoris, Pichia ciferrii, Yarrowia lipolytica, Candida albicans, Candida utilisAshbya gossypii.. The genetically modified cell of or claim 3 , selected from and6. (canceled)7. A method for the production of sphingoid bases and/or sphingolipids claim 3 , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00003', 'claim 3'}, 'a) contacting the genetically modified cell of with a medium containing a carbon source,'}b) culturing the cell under conditions which enable the cell to form sphingoid bases and/or sphingolipids from the carbon source andc) optionally isolating the sphingoid bases and/or sphingolipids formed.8. A method for the production of N-acetylated claim 3 , primary aliphatic amines claim 3 , comprising the steps of: [{'sub': '1', 'an enzyme Ewith a polypeptide sequence in which up to % ...

PROCESS FOR PRODUCING A POLYACRYLAMIDE SOLUTION WITH INCREASED VISCOSITY

The present invention relates to a method for producing a polyacrylamide solution having increased viscosity. In particular, the present invention is related to the separation of a biocatalyst from an aqueous acrylamide solution prepared utilizing the biocatalyst prior to polymerization of the aqueous acrylamide solution to polyacrylamide. A polyacrylamide solution having in creased viscosity is well suited to be used in tertiary oil recovery. Accordingly, the present application provides means and methods to crucially improve the quality of polyacrylamide solutions for use in tertiary oil recovery. 1: A method for producing a polyacrylamide solution having an increased viscosity relative to a reference solution , the method comprising:(a) preparing an aqueous acrylamide solution by converting acrylonitrile to acrylamide using a biocatalyst,(b) separating the biocatalyst from the aqueous acrylamide solution of (a) to obtain an aqueous acrylamide solution having an optical density at 600 nm equal to or less than 0.6, and(c) polymerizing the aqueous acrylamide solution obtained in (b) to obtain a polyacrylamide,wherein the reference solution is a polyacrylamide solution prepared from an aqueous acrylamide solution having an optical density at 600 nm of more than 0.6 and wherein the reference solution is prepared by the same method without separating the biocatalyst.2: A method for preparing a polyacrylamide solution , the method comprising:(a) preparing an aqueous acrylamide solution by converting acrylonitrile to acrylamide using a biocatalyst,{'sup': '2', '(b) separating the biocatalyst from the aqueous acrylamide solution of (a) by disc stack separation performed with a specific settling area of 19.67 mh/l or more to obtain an aqueous acrylamide solution, and'}(c) polymerizing the aqueous acrylamide solution obtained in (b) to obtain a polyacrylamide.3: The method according to claim 1 , further comprising at least one of:(d) drying the polyacrylamide;(e) shredding ...

IMMOBILIZED PROTEINS AND USE THEREOF

The invention relates to an immobilized protein material comprising a protein that is immobilized on a glass material or organic polymer through affinity tag binding. The glass material may be a porous glass material such as (hybrid) controlled porosity glass. The invention also relates to the use of an immobilized enzyme material as a heterogeneous biocatalyst in chemical synthesis. The invention further relates to a method for the immobilization of affinity tagged proteins on a glass material or organic polymer, and to a method for the purification and isolation of affinity tagged proteins by the immobilization of such proteins on a glass material or organic polymer. 1. A method for catalyzing an enzyme-catalyzed cascade reaction , comprising(a) providing a solid support and two or more enzymes immobilized on said solid support,wherein the solid support has pores with diameter ranging between about 10 and 300 nm,wherein the solid support comprises non-swelling carrier material,wherein the non-swelling carrier material comprises a chelated metal ion, and wherein each of said immobilized enzymes comprises an affinity tag that binds to the chelated metal ion; and(b) bringing said immobilized enzymes into contact with a continuous flow of at least one substrate, thereby catalyzing the cascade reaction.2. The method according to claim 1 , wherein at least three enzymes are immobilized on said solid support.3. The method according to claim 1 , wherein the non-swelling carrier material is a porous organic polymer.4. The method according to claim 3 , wherein the porous organic polymer is chosen from the group consisting of polyethylene claim 3 , ultra-high molecular weight polyethylene (UHMWPE) claim 3 , high-density polyethylene (HDPE) claim 3 , polypropylene (PP) claim 3 , polytetrafluoroethylene (PTFE) claim 3 , polyvinylidene fluoride (PVDF) claim 3 , polystyrene claim 3 , polymethacrylate and poly(methyl methacrylate).5. The method according to claim 3 , wherein the ...

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO: 136 , and one or more amino acid substitutions compared to the naturally occurring polypeptide at one or more positions corresponding to positions in SEQ ID NO: 136 , selected from the group consisting of 75 , 79 , 82 , 99 , 110 , 166 , 172 , 208 , 216 , 273 , 324 , 364 , 395 , 412 , 491 , 503 , and 504.3. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 136 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO: 136.5. The non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 136 claim 1 , wherein said polypeptide further comprises one or more substitutions corresponding to substitutions in SEQ ID NO: 136 selected from the group consisting of a glycine at position 143 claim 1 , glycine at position 278 claim 1 , an arginine at position 326 claim 1 , and ...

PICHIA CIFERRII CELLS AND USE THEREOF

The invention relates to genetically modified cells, to the use thereof and to a method of producing sphingoid bases and sphingolipids. 1Pichia ciferrii. An isolated cell , characterized in that the cell has , compared to its wild type , a reduced activity of at least one of the enzymes which are encoded by the intron-free nucleic acid sequences selected from the two groups A) and B) consisting ofA) Seq ID No 1, Seq ID No 3, Seq ID No 5, Seq ID No 7, Seq ID No 9, Seq ID No 11,B) a sequence which is at least 80% identical to any of the sequences Seq ID No 1, Seq ID No 3, Seq ID No 5, Seq ID No 7, Seq ID No 9, Seq ID No 11.2Pichia ciferrii. An isolated cell according to , characterized in that reduction of the enzymic activity is achieved by modifying a gene comprising any of the nucleic acid sequences specified in , the modification being selected from the group comprisinginsertion of foreign DNA into the gene, deletion of at least parts of the gene, point mutations in the gene sequence, exposing the gene to the influence of RNA interference, and replacement of parts of the gene with foreign DNA.3Pichia ciferrii. An isolated cell according to claim 2 , characterized in that the foreign DNA is a selection marker gene claim 2 , preferably one which can be removed without leaving a trace and which leaves a deletion in the target gene.4Pichia ciferriiPichia ciferrii. An isolated cell according to claim 1 , characterized in that the cell derives from strains selected from the group consisting of{'i': 'Pichia ciferrii', 'NRRL Y-1031 F-60-10,'}{'i': 'Pichia ciferrii', 'strains disclosed in WO 95/12683'}{'i': 'Pichia ciferrii', 'CS.PCΔPro2.'}5Pichia ciferrii. An isolated cell according to claim 1 , characterized in that the cell has claim 1 , compared to its wild type claim 1 , an increased enzymic activity of at least one of the enzymes selected from{'sub': '1', 'an enzyme E, which catalyses the reaction of serine and palmitoyl-CoA to give 3-ketosphinganine,'}{'sub': '2', ' ...

Methods Of Producing 6-Carbon Chemicals Via Methyl-Ester Shielded Carbon Chain Elongation

This document describes biochemical pathways for producing adipic acid, 6-aminohexanoic acid, 6-hydroxhexanoic acid, hexamethylenediamine, caprolactam, or 1,6-hexanediol by forming one or two terminal functional groups, comprised of carboxyl, amine or hydroxyl group, in a C6 aliphatic backbone substrate. These pathways, metabolic engineering and cultivation strategies described herein rely on the enzymes or homologs accepting methyl ester shielded dicarboxylic acid substrates. 1. A method for biosynthesizing a product selected from the group consisting of adipic acid , 6-aminohexanoate , 6-hydroxhexanoate , hexamethylenediamine , caprolactam and 1 ,6-hexanediol , said method comprising enzymatically synthesizing a six carbon chain aliphatic backbone from oxalyl-CoA and either (i) acetyl-CoA or malonyl-CoA via two cycles of methyl ester shielded carbon chain elongation or (ii) malonyl-[acp] via two cycles of methyl-ester shielded carbon chain elongation , and enzymatically forming two terminal functional groups selected from the group consisting of carboxyl , amine , and hydroxyl groups in said backbone , thereby forming the product.2. The method of claim 1 , wherein the six carbon chain aliphatic backbone is adipyl-[acp] or adipyl-CoA.3. The method of claim 1 , wherein a malonyl-[acp] O-methyltransferase converts oxalyl-CoA to oxalyl-CoA methyl ester.4. The method of claim 3 , wherein each of said two cycles of carbon chain elongation comprises using (i) a β-ketoacyl-[acp] synthase or a β-ketothiolase claim 3 , (ii) a 3-oxoacyl-[acp] reductase claim 3 , an acetoacetyl-CoA reductase claim 3 , a 3-hydroxyacyl-CoA dehydrogenase or a 3-hydroxybutyryl-CoA dehydrogenase claim 3 , (iii) an enoyl-CoA hydratase or a 3-hydroxyacyl-[acp] dehydratase claim 3 , and (iv) an enoyl-[acp] reductase or a trans-2-enoyl-CoA reductase to produce adipyl-[acp] methyl ester or adipyl-[acp] methyl ester.5. The method of claim 4 , wherein a pimeloyl-[acp] methyl ester methylesterase removes ...

METHOD FOR PRODUCING COMPOUND AND COMPOUND PRODUCTION SYSTEM USED IN PRODUCTION METHOD

This method for producing a compound uses a continuous tank reactor which is provided with two or more reaction tanks for producing the compound and with a reaction liquid feeding pipe that feeds a reaction liquid from an upstream reaction tank to a downstream reaction tank, said method being characterized in that the Reynold's number of the reaction liquid that flows in the reaction liquid feeding pipe is configured to be 1800-22000. Furthermore, this compound production system is used in said method for producing a compound, and is formed by housing at least one of the reaction tanks in a portable container. 1. A method for producing a compound in a continuous tank reactor , the method comprising:feeding a reaction liquid from an upstream reaction tank to a downstream reaction tank, via a reaction liquid feeding pipe ata Reynolds number of 1800 to 22000,wherein the continuous tank reactor comprises two or more reaction tanks and the reaction liquid feeding pipe.2. A compound production system for the method according to claim 1 , wherein at least one tank of the reaction tanks is accommodated in a portable container.3. The compound production system according to claim 2 , whereina total volume of the at least one tank of the reaction tanks accommodated in the portable container is ⅙ to ⅗ of an inner volume of the portable container.4. The method according to claim 1 , wherein at least one tank of the reaction tanks is accommodated in a portable container claim 1 , and a value obtained by dividing an inner volume (m) of the portable container by a flow rate (m/hour) of the reaction liquid flowing in the reaction tank accommodated in the portable container is 5 to 70 hours.5. The compound production system according to claim 2 , wherein another portable container is disposed on the portable container in which the reaction tank is accommodated.6. A process for producing acrylamide using the method according to claim 1 , the process comprising:supplying an ...

Human microbiota derived N-acyl amides for the treatment of human disease

The present invention provides compositions and methods for the modulation of G protein-coupled receptors (GPCRs).

ANTIMICROBIAL AGENTS

The invention provides novel compounds of formula (I) and their pharmaceutically acceptable salts, metabolites, isomers (e.g. stereoisomers) and prodrugs. Such compounds are effective in the treatment of infections caused by Gram-negative bacteria such as . In formula (I), X is O, NR (where R is either H or Calkyl, e.g. CH), or CH; Ris H, F, CI, Br, I, or CH; Ris H, or OH; Rand Rare independently selected from H and OH, or Rand Rtogether are ═O; Ris H, F, CI, Br, I, or CH; Ris H, OH, or —OC(O)NR′(where each R′ is independently H or Calkyl, e.g. CH), preferably Ris H, OH or —OC(O)NH; Ris a 5- or 6-membered, saturated or unsaturated, carbocyclic ring optionally substituted by one or more substituents, or Ris an optionally substituted straight-chained or branched Calkyl group (e.g. Calkyl group); Ris a straight-chained or branched Calkyl group (e.g. Calkyl group), a Ccycloalkyl group, or an optionally substituted aryl or heteroaryl group; and each --- independently represents an optional bond (i.e. each of C-C, C-C, C-C, C-C, C-Cand C-Care independently either C—C (single) or C═C (double) bonds). 2. A compound as claimed in claim 1 , wherein Ris an optionally substituted cyclohexyl or cyclopentyl ring claim 1 , an optionally substituted cyclohexenyl ring claim 1 , or an optionally substituted claim 1 , straight-chained Calkyl group.3. A compound as claimed in or claim 1 , wherein Ris substituted by one or more of the following groups: OH claim 1 , NR(where each Ris independently H or Calkyl claim 1 , e.g. CH) claim 1 , SR(where Ris H or Calkyl claim 1 , e.g. CH) claim 1 , halogen (e.g. F claim 1 , Cl claim 1 , Br claim 1 , or I) claim 1 , Calkyl (e.g. CH) claim 1 , COH (or an ester thereof) claim 1 , POH(or an ester thereof) and SOH(or an ester thereof).4. A compound as claimed in any one of to claim 1 , wherein Ris a straight-chained or branched Calkyl (e.g. Calkyl) group claim 1 , preferably a straight-chained or branched Calkyl claim 1 , more preferably a straight- ...

Biotechnological Method for the Production of Acrylamide and Relative New Bacterial Strain

A bacterial strain of named Palladio 22 and registered at the BCCM-LMG Bacteria Collection under registration number LMG P-29520. A method is provided for the production of acrylamide following hydration of acrylonitrile using a biomass of the bacterial strain. 1Rhodococcus biphenylivorans. A bacterial strain of named Palladio 22 and registered at the BCCM-LMG Bacteria Collection under registration number LMG P-29520.2. The bacterial strain according to claim 1 , which produces a hydratase nitrile enzyme.3. The bacterial strain according to claim 1 , in the form of dried claim 1 , lyophilized or paste biomass having a dry residue from 18 to 30%.4. Use of the bacterial strain according to for the production of an amide.5. The use according to claim 4 , wherein the amide is acrylamide.6. A method for the production of acrylamide by hydration of acrylonitrile using a biomass of the bacterial strain according to claim 1 , wherein the hydration process takes place at a concentration of acrylonitrile smaller than 0.8% of the total weight of the reaction solution.7. The method according to claim 6 , wherein acrylonitrile is added to an aqueous solution containing the biomass of the bacterial strain so that claim 6 , during the hydration process claim 6 , the concentration of free acrylonitrile in solution is never greater than 0.8% of the total weight of the reaction solution.8. The method according to claim 6 , having an acrylamide production yield of between 50 and 57.5% (weight/weight).9. The method according to claim 8 , having an acrylamide production yield of between 50 and 54%.10. The method according to claim 6 , wherein the production of acrylamide takes place in a temperature range of between 14 and 23° C. and pH of between 5.0 and 8.5.11. A method for the production of acrylamide by hydration of acrylonitrile using a biomass of the bacterial strain according to claim 1 , wherein said biomass is immobilized on a solid substrate.12. A method for the production of ...

Yeast Strain with High Yield of Glutathione

The present invention discloses a yeast strain with high yield of glutathione, and belongs to the technical field of microorganisms. tlj2016 provided in the present invention has been deposited in China General Microbiological Culture Collection Center of Microbe Preservation Management Committee, with a preservation number of CGMCC No. 12789. The strain can tolerate glucose with tolerance capability reaching 300 g/L, and has tolerance capability against L-cysteine that is far higher than that of the starting strain, contributing to the production of glutathione at conditions of high concentrations of glucose, with a final concentration of GSH produced through fermentation in a 5-L fermentation tank reaching 3308 mg/L. 1Saccharomyces cerevisiaeSaccharomyces cerevisiaeSaccharomyces cerevisiae. A strain of , characterized in that , the is specifically tlj2016 with accession number of CGMCC No. 12789.2. (canceled) The present invention belongs to the technical field of microorganisms, and particularly relates to a yeast strain with high yield of glutathione.Glutathione (GSH) is a biologically active tripeptide compound formed by condensation of L-glutaminic acid, L-cysteine and glycine, and is widely present in animal, plant and microbial cells. GSH has a variety of important physiological functions within organisms, and particularly plays an important role in the maintenance of a suitable oxidation-reduction environment within the organisms, so that it has wide applications in clinical, food and cosmetic industries.Methods for producing GSH mainly include chemical synthesis methods, enzyme methods, fermentation methods and the like. In the chemical synthesis methods, three precursor amino acids are used as raw materials to carry out chemical synthesis, but it is not easy to separate the resulting active products and thus the product purity is not high, so that the application of these methods is limited. In the enzyme synthesis, three substrate amino acids and GSH ...

IMPROVED NITRILE HYDRATASE

Provided is an improved nitrile hydratase with improved catalytic activity. Also provided are DNA for coding the improved nitrile hydratase, a recombinant vector that contains the DNA, a transformant that contains the recombinant vector, nitrile hydratase acquired from a culture of the transformant, and a method for producing the nitrile hydratase. Also provided is a method for producing an amide compound that uses the culture or a processed product of the culture. The improved nitrile hydratase contains an amino acid sequence represented by SEQ ID NO: 50 (GXXXXDXXR) in a beta subunit, and is characterized in that Xis an amino acid selected from a group comprising cysteine, aspartic acid, glutamic acid, histidine, isoleucine, lysine, methionine, asparagine, proline, glutamine, serine and threonine. 2. (canceled)3. The improved nitrile hydratase according to claim 1 , wherein Xis I (isoleucine) claim 1 , Xis S (serine) claim 1 , Xis W (tryptophan) claim 1 , Xis K (lysine) claim 1 , and Xis S (serine) in SEQ ID NO: 50.4. The improved nitrile hydratase according to claim 1 , further comprising an amino-acid sequence in SEQ ID NO: 51 comprising the amino-acid sequence in SEQ ID NO: 50.5. (canceled)6. The improved nitrile hydratase according to claim 1 , wherein Xis G (glycine) claim 1 , Xis R (arginine) claim 1 , Xis T (threonine) claim 1 , Xis L (leucine) claim 1 , Xis S (serine) claim 1 , Xis I (isoleucine) claim 1 , Xis T (threonine) claim 1 , Xis W (tryptophan) claim 1 , Xis M (methionine) claim 1 , Xis H (histidine) claim 1 , Xis L (leucine) claim 1 , Xis K (lysine) claim 1 , and Xis G (glycine) in SEQ ID NO: 81.7. The improved nitrile hydratase according to claim 1 , further comprising an amino-acid sequence in SEQ ID NO: 82 comprising the amino-acid sequence in SEQ ID NO: 81.8. The improved nitrile hydratase according to claim 1 , wherein Xis M (methionine) claim 1 , Xis A (alanine) claim 1 , Xis S (serine) claim 1 , Xis L (leucine) claim 1 , Xis Y (tyrosine) ...

Novel urethanases for the enzymatic degradation of polyurethanes

The present invention relates to new urethanases for the enzymatic breakdown of polyurethanes and to an enzymatic process for the complete breakdown of polyurethanes into defined monomers.

NOVEL ASPARTOKINASE VARIANT AND METHOD FOR PRODUCING L-AMINO ACID USING THE SAME

An aspartokinase variant, a microorganism comprising the variant, and a method for producing an aspartate-derived L-amino acid or a homoserine derivative thereof using the microorganism. 1. An aspartokinase variant , wherein the amino acid at position 377 in the amino acid sequence of SEQ ID NO: 1 is substituted with L-lysine or L-methionine.2. A polynucleotide encoding the variant of .3Corynebacterium. A microorganism of the genus claim 1 , which produces an aspartate-derived L-amino acid or an amino acid derivative thereof comprising the aspartokinase variant of claim 1 , wherein the aspartate-derived L-amino acid or amino acid derivative thereof is selected from the group consisting of lysine claim 1 , threonine claim 1 , methionine claim 1 , homoserine claim 1 , O-acetylhomoserine claim 1 , O-succinylhomoserine claim 1 , isoleucine claim 1 , and cadaverine.4. (canceled)5CorynebacteriumCorynebacterium glutamicum.. The microorganism according to claim 3 , wherein the microorganism of the genus is6. A method for producing an aspartate-derived L-amino acid or an amino acid derivative thereof claim 3 , comprising:{'claim-ref': {'@idref': 'CLM-00003', 'claim 3'}, 'culturing the microorganism of in a medium; and'}recovering the aspartate-derived L-amino acid or amino acid derivative thereof from the cultured microorganism or cultured medium, wherein the aspartate-derived L-amino acid or amino acid derivative thereof is selected from the group consisting of lysine, threonine, methionine, homoserine, O-acetylhomoserine, O-succinylhomoserine, isoleucine, and cadaverine.7. (canceled)8CorynebacteriumCorynebacterium glutamicum.. The method according to claim 6 , wherein the microorganism of the genus is9. A method for producing L-methionine claim 6 , comprising:{'claim-ref': {'@idref': 'CLM-00003', 'claim 3'}, 'culturing the microorganism of in a medium;'}producing O-acetylhomoserine or O-succinylhomoserine from the cultured microorganism or cultured medium; andconverting ...

COMPOSITIONS AND METHODS FOR MICROBIAL CO-CULTURE

Provided herein are compositions and method for producing γ-polyglutamic acid (PGA). In particular, provided herein are bacterial co-culture systems and methods for producing PGA. 1. A method of producing γ-polyglutamic acid (PGA) , comprising:a) contacting a fungus with a first feedstock;b) fermenting said fungus to generate citric acid;{'i': 'Bacillus', 'c) contacting said citric acid from step b) with a sp. and a second feedstock; and'}{'i': 'Bacillus', 'd) fermenting said sp. to generate PGA.'}2. The method of claim 1 , wherein one or more of steps a-d) further comprises the addition of one or more of glycerol claim 1 , glutamate claim 1 , or glutamine.3. The method of claim 1 , wherein steps b) and d) are conducted in the same or different bioreactors.4Aspergillus niger, Yarrowia lipolyticaCandida oleophila.. The method of claim 1 , wherein said fungus is claim 1 , or5BacillusBacillus subtilisBacillus licheniformis.. The method of claim 1 , wherein said sp. is or6. The method of claim 5 , wherein said Bacillus subtilis is a strain selected from the group consisting of IFO 3335 claim 5 ,TAM-4 claim 5 , C1 claim 5 , C10 claim 5 , chungkookjang claim 5 , NX-2 claim 5 , MR-141 claim 5 , CGMCC 0833 claim 5 , R23 claim 5 , ER1001 claim 5 , ER1007 claim 5 , and ER1012 claim 5 , and RKY3.7Bacillus licheniformis. The method of claim 5 , wherein said is a strain selected from the group consisting of SAB-26 claim 5 , A35 claim 5 , ATCC 9945 claim 5 , CC 12826 claim 5 , WBL-3 claim 5 , and NCIM 2324.8. The method of claim 1 , wherein said citric acid is isolated after said step b).9. The method of claim 1 , wherein said first and second feedstocks are selected from the group consisting of molasses claim 1 , raffinate claim 1 , pomace claim 1 , fruit peels claim 1 , corn starch claim 1 , wheat starch claim 1 , sorghum claim 1 , brewery wastes claim 1 , corn stover claim 1 , spent algae cake claim 1 , and glycerol.10. The method of claim 9 , wherein said first and second ...

APPLICATION OF GLUTAMATE DEHYDROGENASE GDHA OF PEPTOSTREPTOCOCCUS ASACCHAROLYTICUS IN INCREASING YIELD OF POLY- r -GLUTAMIC ACID FROM BACILLUS LICHENIFORMIS

Application of glutamate dehydrogenase GdhA of in increasing the yield of poly-γ-glutamic acid from . The glutamate dehydrogenase GdhA of the WX-02 per se is replaced with the glutamate dehydrogenase derived from the by means of homologous recombination, which significantly increases the level of synthesizing the poly-γ-glutamic acid for the , and the yield of the obtained poly-γ-glutamic acid from strains is increased at least by more than 20% compared with control strains. 1Bacillus licheniformis. A method for increasing yield of poly-γ-glutamic acid produced by through fermentation , comprising:{'i': Bacillus licheniformis', 'Peptostreptococcus asaccharolyticus, 'replacing a glutamate dehydrogenase gene in with a glutamate dehydrogenase gene of to obtain a recombinant strain,'}producing the poly-γ-glutamic acid through fermentation of the recombinant strain.2Bacillus licheniformisBacillus licheniformis. The method according to claim 1 , wherein the is WX-02 with CCTCC Accession No. M208065.3Peptostreptococcus asaccharolyticus. The method according to claim 1 , wherein the glutamate dehydrogenase gene of the is as shown in SEQ ID NO:2.4. The method according to claim 1 , wherein fermentation media used during the fermentation comprises:{'sub': 3', '4', '2', '4', '2', '4', '2', '4', '2', '4', '2', '2, '30-90 g/L of glucose, 0-30 g/L of sodium glutamate, 0-10 g/L of sodium citrate, 5-10 g/L of NaNO, 0-10 g/L of NHCl, 0.5-1 g/L of KHPO.3HO, 0.8-1.2 g/L of MgSO.7HO, 0.8-1.2 g/L of ZnSO.7HO, 0.1-0.2 g/L of MnSO—HO, and 0.8-1.2 g/L of CaCl, wherein at most one of the sodium glutamate, the sodium citrate, and the ammonium chloride is optionally 0 in the content; or'}{'sub': 3', '4', '2', '4', '2', '4', '2', '4', '2', '4', '2', '2, '18-22 g/L of glycerol, 25-35 g/L of sodium glutamate, 8-13 g/L of sodium citrate, 7-12 g/L of NaNO, 8-12 g/L of NHCl, 0.8-1.2 g/L of KHPO.3HO, 0.9-1.2 g/L of MgSO.7HO, 0.8-1.2 g/L of ZnSO.7HO, 0.1-0.25 g/L of MnSO.HO, and 0.5-1.5 g/L of CaCl ...

Synthetic spilanthol and use thereof

The present application provides a synthetic spilanthol flavor composition that includes (2E,6Z,8E)-N-(2-methylpropyl)-2,6,8-decatrienamide. The synthetic spilanthol composition can also contain, and at least one of (2E,6E,8E)-N-(2-methylpropyl)-2,6,8-decatrienamide and (2E,6Z,8Z)—N-(2-methylpropyl)-2,6,8-decatrienamide, the (N-(2-methylpropyl)-2,6,8-decatrienamide being present in amounts effective to impart a salivating or tingle effect while reducing the perception of off notes, as compared to the off-notes perceived upon consumption of natural spilanthol (e.g. spilanthol obtained from jambu oleoresin). Methods of increasing salivation and/or providing a tingling sensation upon consuming an orally consumable product that include adding to the product a synthetic spilanthol flavor composition are also provided. Synthetic spilanthol flavor compositions can be added to orally consumed products, such as, but not limited to, foods, beverages, pharmaceuticals, nutraceuticals, or therapeutic compositions, oral personal care products, gums (e.g. chewing gum or bubble gum), candy or lozenges.

Method for preparing an aqueous polyacrylamide solution

A method for preparing an aqueous polyacrylamide solution is disclosed. The method comprises:—hydrating acrylonitrile in water in presence of a biocatalyst capable of converting acrylonitrile to acrylamide so as to obtain an acrylamide solution,—directly polymerizing the acrylamide solution so as to obtain a polyacrylamide gel, and—directly dissolving the polyacrylamide gel by addition of water so as to obtain an aqueous polyacrylamide solution.

Polypeptides Having Peroxygenase Activity and Polynucleotides Encoding Same

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. 129-. (canceled)30. An isolated polypeptide having peroxygenase activity , selected from the group consisting of:(a) a polypeptide having at least 60% sequence identity to the mature polypeptide of SEQ ID NO: 2;(b) a polypeptide encoded by a polynucleotide that hybridizes under medium stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii);(c) a polypeptide encoded by a polynucleotide having at least 60% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1, or the cDNA sequence thereof;(d) a variant of the mature polypeptide of SEQ ID NO: 2 comprising a substitution, deletion, and/or insertion at one or more positions; and(e) a fragment of the polypeptide of (a), (b), (c), or (d) that has peroxygenase activity.31. A composition comprising the polypeptide of .32. A detergent composition claim 30 , comprising a surfactant and a polypeptide according to .33. A method for hydroxylation in position 2 or 3 of either end of a substituted or unsubstituted claim 30 , linear or branched claim 30 , aliphatic hydrocarbon having at least 3 carbons and having a hydrogen attached to the carbon in position 2 or 3 claim 30 , comprising contacting the aliphatic hydrocarbon with hydrogen peroxide and a polypeptide according to .34. A method for hydroxylation in position 2 or 3 of the terminal end of an acyl group of a lipid claim 30 , comprising contacting the lipid with hydrogen peroxide and a polypeptide according to .35. A method for introducing a hydroxy or a keto group at the second or third carbon of at least two ends of a substituted or ...

Recombinant strain producing o-aminobenzoate and fermentative production of aniline from renewable resources via 2-aminobenzoic acid

The invention provides a recombinant microbial host cell capable of converting a raw material comprising a fermentable carbon substrate to o-aminobenzoate biologically. The invention further provides a method for producing aniline, comprising the steps of: a) producing o-aminobenzoate by fermentation of a raw material comprising at least one fermentable carbon substrate using the recombinant microbial host cell of the capable of converting said raw material comprising at least one fermentable carbon substrate to o-aminobenzoate biologically, wherein said o-aminobenzoate comprises anthranilate anion, b) converting said o-aminobenzoate from said anthranilate anion to anthranilic acid by acid protonation, c) recovering said anthranilic acid by precipitation or by dissolving in an organic solvent, and d) converting said anthranilic acid to aniline by thermal decarboxylation in an organic solvent.

METHOD FOR PRODUCING PULVERULENT CERAMIDE

Disclosed herein is a pulverulent ceramide producing method that enables easily and efficiently extracting and separating ceramide from sugar beet pulp, and efficiently pulverizing the ceramide by spray drying. The pulverulent ceramide can be efficiently obtained by a process that includes concentrating, with and/or without adding water, a sugar beet pulp ethanol extract obtained by extraction of a sugar beet pulp (for example, such as a beet fiber) with ethanol, adding pectinase to the resulting concentrate and performing an enzymatic reaction, performing emulsification after inactivating the enzyme, and pulverizing the resulting emulsion using spray drying. 1. A method for producing pulverulent ceramide ,the method comprising:concentrating, with and/or without adding water, a sugar beet pulp ethanol extract obtained through extraction of a sugar beet pulp with ethanol to obtain a concentrate;adding pectinase to thus obtained concentrate thus performing an enzymatic reaction to degrade pectin;inactivating the pectinase and emulsifying the product of the enzymatic reaction to produce an emulsion; andspray drying the resulting emulsion to produce a pulverant ceramide.2. The method according to claim 1 , wherein the enzymatic reaction is performed at a temperature of 10 to 70° C. for 6 minutes or more.3. The method according to claim 1 , wherein the enzymatic reaction is performed at a temperature of 45 to 50° C. for 0.5 to 2 hours.4. The method according to claim 1 , wherein the pectinase is added in 0.0002 weight % or more of an amount of the raw material sugar beet pulp.5. The method according to claim 1 , wherein the pectinase is added in 0.005 to 0.2 weight % of an amount of the raw material sugar beet pulp.6. The method according to claim 1 , wherein an excipient is added as a pulverization auxiliary agent before the emulsification.7. The method according to claim 6 , wherein a processed starch is added as the excipient.8. A method for improving a spray drying ...

FUSION PROTEINS USEFUL LN THE PRODUCTION OF CANNABINOIDS

The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular/modified/synthetic gene(s) of select enzymes are processed and inserted into an expression system (vector, cosmid, BAC, YAC, phage, etc.) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product. 113-. (canceled)14. An isolated codon-optimized nucleic acid sequence of SEQ ID NO: 34 or a nucleotide sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 34: or an isolated codon-optimized nucleic acid sequence of nucleic acids 5841 to 8867 of SEQ ID NO: 34 or a nucleotide sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acids 5841 to 8867 of SEQ ID NO: 34.15. An isolated codon-optimized nucleic acid sequence of SEQ ID NO: 35 or a nucleic acid sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acid sequence of SEQ ID NO: 35; or an isolated codon-optimized nucleic acid sequence of nucleic acids 5052 to 6671 of SEQ ID NO: 35 or a nucleotide sequence that is 95% or more , 96% or more , 97% or more , 98% or more , or 99% or more homologous to the nucleic acids 5052 to 6671 of SEQ ID NO: 35.16. An isolated ...

Process for producing acyl amino acids employing lipases

There is provided a microbial cell for producing at least one acyl amino acid, wherein the cell is genetically modified to comprise: a first genetic mutation that increases the expression relative to a wild type cell of at least one lipase (EC 3.1.1) (E 1 ) capable of hydrolysing at least one glyceride to at least one fatty acid wherein the glyceride is a triglyceride; and a second and a third genetic mutation that increases the expression relative to a wild type cell of: (i) an amino acid-N-acyl-transferase (EC 2.3.1) (E 2 ), and (ii) an acyl-CoA synthetase (EC 6.2.1.3) (E 3 ) respectively that enables the cell to convert the fatty acid to at least one acyl amino acid and wherein the cell has a reduced fatty acid degradation capacity.

Method for Vegetable Oil Deacidification by Enzymatic Amidation

The present invention provides a method for vegetable oil deacidification by enzymatic amidation, which relates to the field of oil refining technology. The present invention is carried out through mixing high acid value vegetable oil with ethanolamine at a certain molar ratio in a solvent or solvent-free system, adding with a certain amount of lipase, and reacting at certain temperature for a period of time. The monoethanolamine has been used as an acyl donor for the first time to react with free fatty acid, which avoids the increasing amount of by-products and great loss of neutral oil in reaction that involved with triglycerides. The method of the present invention has the advantages of high selectivity, high catalytic efficiency, and environment friendly in the reaction. From enzymes recycling, it greatly reduces costs, which shows tremendous potential in the application. 1. A method for vegetable oil deacidification by enzymatic amidation , wherein said method comprises mixing high acid value vegetable oil with ethanolamine at a certain molar ratio in a solvent or solvent-free system , adding with certain amount of lipase , and reacting at certain condition; wherein said molar ratio is 1:(1-3) for free fatty acids and monoethanolamine; wherein said lipase is at an amount of 1˜10% of total quality of high acid oil and monoethanolamine; and wherein said condition is at 50˜100 for 1˜35 h.2. The method of claim 1 , wherein said solvent system comprises one or more compounds selected from a group consisting of hexane claim 1 , petroleum ether claim 1 , ethyl acetate claim 1 , diethyl ether claim 1 , acetonein and a combination thereof.3. The method of claim 2 , the high acid value vegetable oil and organic solvent is at a ratio of 1:(0.5-5) (g/mL).4. The method of claim 1 , wherein said system is solvent-free system claim 1 , and the reaction is under 0.075˜0.1 Mpa.5. The method of claim 1 , wherein said lipase comprises one or more immobilized enzymes selected from ...

IN VIVO AND IN VITRO OLEFIN CYCLOPROPANATION CATALYZED BY HEME ENZYMES

The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention. 1. A reaction mixture for producing a cyclopropanation product , the reaction mixture comprising an olefinic substrate , a carbene precursor , and a heme enzyme.3. The reaction mixture of claim 2 , wherein:{'sup': 1', 'c, 'Ris C(O)O-LR;'}{'sup': '2', 'sub': '6-10', 'Ris selected from the group consisting of H and optionally substituted Caryl; and'}{'sup': 3', '4', '5', '6, 'sub': 1-6', '2-6', '2-6', '6-10, 'R, R, R, and Rare independently selected from the group consisting of H, optionally substituted Calkyl, optionally substituted Calkenyl, optionally substituted Calkynyl, optionally substituted Caryl and halo,'}{'sup': 3', '4, 'or Rforms an optionally substituted 3- to 18-membered ring with R;'}{'sup': 5', '6, 'or Rforms an optionally substituted 3- to 18-membered ring with R.'}10. The reaction mixture of claim 1 , wherein the cyclopropanation product is an intermediate of milnacipran claim 1 , levomilnacipran claim 1 , cilastain claim 1 , or sitafloxacin.12. The reaction mixture of claim 1 , wherein the olefinic substrate is selected from the group consisting of an alkene claim 1 , a cycloalkene claim 1 , and an arylalkene.13. The reaction mixture of claim 12 , wherein the arylalkene is a styrene.16. The reaction mixture of ...

CALB Variants

The invention relates to amino acid sequence variants of a lipase with improved activity for catalyzing synthesis reactions and methods of preparing the variants. The methods include predicting amino acid sites for change based on computational models of the protein structure in non-aqueous conditions, and expressing the protein in a prokaryotic host for subsequent purification and use. The enzyme sequence variants described have a three to nine-fold improvement in synthesis activity over the parent protein sequence.

Manumycin-type metabolite called Colabomycin E which inhibits caspase 1 and creation of interleukins, strain produces the Colabomycin E and a method of a production of the Colabomycin E

A manumycin-type metabolite called Colabomycin E which inhibits caspase 1 and creation of interleukins, strain produces the Colabomycin E and a method of a production of the Colabomycin E. Colabomycin E is a new member of the manumycin-type metabolites produced by the strain Streptomyces aureus SOK1/5-04 deposited in The Czech collection of microorganisms under number CCM8556. The structure of 5 is similar to that of the already known metabolite colabomycin A (3) isolated from Streptomyces griseoflavus . However, the upper polyene chain of 5 is two carbons longer. Therefore, it was named Colabomycin E. Biological activity assays indicated that colabomycin E significantly inhibited IL-1β release from THP-1 cells and might thus potentially act as an anti-inflammatory agent.

Aspartase variants, method of preparing the same and use thereof

The present invention discloses an aspartase variant, method of preparing the same and use thereof. Compared with sequence 2 in the sequence listing, the amino acid sequence of the aspartase variant provided in the present invention has more than 96% identity, and there are mutations in T187I and N326C at positions 187 and 326, respectively, and has improved catalytic activity for the ammoniation of acrylic acid, compared with the aspartase shown in sequence 2. The experiment proves that the aspartase variant provided in the present invention does have improved catalytic activity for the ammoniation of acrylic acid compared with the wild type parent aspartase, and has better thermal stability and pH spectrum. The reaction can greatly increase the conversion ratio of acrylic acid.

PROCESS FOR THE PRODUCTION OF GAMMA-AMINOBUTYRIC ACID

The present invention relates to a novel method for the fermentative production of gamma-aminobutyric acid (GABA) by cultivating a recombinant microorganism expressing an enzyme having a glutamate decarboxylase activity. The present invention also relates to corresponding recombinant hosts, recombinant vectors, expression cassettes and nucleic acids suitable for preparing such hosts as well as to a method for preparing polyamides making use of GABA as obtained fermentative production.

BIOTECHNOLOGICAL SYNTHESIS PROCESS OF OMEGA-FUNCTIONALIZED CARBON ACIDS AND CARBON ACID ESTERS FROM SIMPLE CARBON SOURCES

The subject of the invention is a biotechnological process for the production of ω-functionalized carboxylic acids and of ω-functionalized carboxylate esters from simple carbon sources.

BIOCATALYSTS AND METHODS FOR THE SYNTHESIS OF ARMODAFINIL

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess. 1. A non-naturally occurring polynucleotide encoding a non-naturally occurring polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 90% sequence identity to SEQ ID NO: 2 , and one or more amino acid substitutions compared to the naturally occurring polypeptide at one or more positions corresponding to positions in SEQ ID NO: 2 , selected from the group consisting of 280 , 322 , 325 , 426 , 430 , 432 , 435 , and 532.3. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid substitutions relative to SEQ ID NO: 2 claim 1 , wherein the polypeptide comprises an alanine claim 1 , glutamic acid claim 1 , glycine claim 1 , isoleucine claim 1 , lysine claim 1 , proline claim 1 , serine claim 1 , threonine claim 1 , or valine at a position corresponding to position 246 of SEQ ID NO: 2.5. The non-naturally occurring polynucleotide encoding the non-naturally occurring polypeptide of claim 1 , wherein said non-naturally occurring polypeptide further comprises one or more amino acid differences relative to SEQ ID NO: 2 claim 1 , wherein said polypeptide further comprises one or more substitutions corresponding to substitutions in SEQ ID NO: 2 selected from the group consisting of a glycine at position ...

EFFICIENT SYNTHESIS OF AMINES AND AMIDES FROM ALCOHOLS AND ALDEHYDES BY USING CASCADE CATALYSIS

The present invention relates generally to an eco-friendly methodology for the conversion of alcohols and aldehydes to amines and amides using an integrated enzyme cascade system with metal- and organocatalysis. More specifically, the present invention relates to synthesis of capsaicinoids starting from vanillin alcohol and using a combination of an enzyme cascade system and catalysts. Furthermore, the method also relates to synthesis of capsaicinoids derivatives starting from vanillin alcohol derivatives and using a combination of an enzyme cascade system and catalysts. 1. A method for conversion of an alcohol comprising:a. Converting said alcohol to an aldehyde or a ketone,b. Converting said aldehyde or ketone to an amine, wherein the conversion of said aldehyde or ketone to amine is catalyzed by an enzyme cascade system, andc. Converting said amine to said amide,wherein said enzyme cascade system comprises Amine Transaminase (ATA).2. The method according to claim 1 , wherein said enzyme cascade system comprises ATA and an amine donor.3. The method according to claim 1 , wherein:said alcohol is a primary alcohol,said aldehyde comprises an R group wherein R is chosen from the group consisting of alkyl, aryl, cinnamyl and heterocyclic,said amine comprises an R group wherein R is chosen from the group consisting of alkyl, aryl, cinnamyl and heterocyclic, and{'sup': 3', '3, 'said amide comprises (i) an R group wherein R is chosen from the group consisting of alkyl, aryl, cinnamyl and heterocyclic, and (ii) an acyl group (COR) wherein Ris chosen from the group consisting of alkyl and aryl.'}4. The method according to claim 1 , wherein:said alcohol is an aldol,said aldehyde comprises an R group wherein R is chosen from the group consisting of alkyl, aryl, cinnamyl and heterocyclic,said amine comprises a R group wherein R is chosen from the group consisting of alkyl, aryl, cinnamyl and heterocyclic, and{'sup': 3', '3, 'said amide comprises (i) an R group wherein R is ...

PACTAMYCIN ANALOGS AND METHODS OF MAKING THEREOF

This disclosure describes the molecular cloning of a pactamycin biosynthetic gene cluster from ATCC 27456, characterization of individual genes in the gene cluster and the proteins encoded thereby as well as uses thereof. The pactamycin gene cluster is located within an 86.35 kilobases genetic locus and includes 53 open reading frames, 26 of which are considered to be the core cluster directly involved in the biosynthesis of pactamycin. The present disclosure also relates to the use of the pactamycin biosynthetic genes located within the identified gene cluster for drug design and development purposes, including the development of pactamycin analogs that are more efficacious and less toxic. Also provided are drugs and antibiotics so produced, as well as methods of their use. 2. The compound of claim 1 , wherein Ris carbamoyl (—C(O)NRR).3. The compound of claim 1 , wherein Ris dimethylcarbamoyl (Rand Rare methyl).4. The compound of claim 1 , wherein Ris lower alkyl.5. The compound of claim 4 , wherein the lower alkyl is methyl claim 4 , ethyl or hydroxyalkyl.6. The compound of claim 5 , wherein the lower alkyl is methyl.7. The compound of claim 1 , wherein Ris lower alkyl.8. The compound of claim 7 , wherein the lower alkyl is methyl.9. The compound of claim 1 , wherein Ris dimethylcarbamoyl and Rand Rare methyl.10. The method of making the compound of claim 1 , comprisingtransforming a host cell with one or more expression vectors comprising an isolated nucleic acid molecule encoding a protein consisting of SEQ ID Nos.: 1 to 53; and{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'culturing the host cell in a culture medium to produce the compound of .'}11. The method of claim 10 , wherein Ris carbamoyl (—C(O)NRR).12. The method of claim 12 , wherein Ris dimethylcarbamoyl (Rand Rare methyl).13. The method of claim 10 , wherein Ris lower alkyl.14. The method of claim 13 , wherein the lower alkyl is methyl claim 13 , ethyl or hydroxyalkyl.15. The method of claim 14 , ...

METHOD FOR PRODUCING SPHINGOID BASE OR SPHINGOLIPID

A method for producing an objective substance such as sphingoid bases and sphingolipids using yeast is provided. An objective substance is produced by cultivating yeast having an ability to produce the objective substance in a culture medium containing an additive that is able to associate with, bind to, solubilize, and/or capture the objective substance, and collecting the objective substance from cells of the yeast and/or the culture medium. 1. A method for producing an objective substance , the method comprising:cultivating yeast having an ability to produce the objective substance in a culture medium containing an additive that is able to associate with, bind to, solubilize, and/or capture the objective substance; andcollecting the objective substance from cells of the yeast and/or the culture medium,wherein the objective substance is selected from the group consisting of sphingoid bases and sphingolipids.2. The method according to claim 1 , wherein the additive is selected from the group consisting of cyclodextrin and zeolite.3. The method according to claim 2 , wherein the cyclodextrin is selected from the group consisting of alpha-cyclodextrin claim 2 , beta-cyclodextrin claim 2 , gamma-cyclodextrin claim 2 , and derivatives thereof.4. The method according to claim 3 , wherein the derivatives are selected from the group consisting of methyl-alpha-cyclodextrin claim 3 , methyl-beta-cyclodextrin claim 3 , hydroxypropyl-alpha-cyclodextrin claim 3 , and hydroxypropyl-beta-cyclodextrin.5. The method according to claim 1 , wherein the objective substance is selected from the group consisting of phytosphingosine (PHS) claim 1 , sphingosine claim 1 , 6-hydroxysphingosine claim 1 , sphinganine (DHS) claim 1 , tetraacetylphytosphingosine (TAPS) claim 1 , triacetylphytosphingosine claim 1 , diacetylphytosphingosine claim 1 , phytoceramides claim 1 , dihydroceramides claim 1 , 6-hydroxyceramides claim 1 , and glucosylceramides.6. The method according to claim 1 , wherein ...

BIOCATALYSTS FOR THE PREPARATION OF HYDROXY SUBSTITUTED CARBAMATES

The present disclosure relates to engineered ketoreductase polypeptides for the preparation of hydroxyl substituted carbamate compounds, and polynucleotides, vectors, host cells, and methods of making and using the ketoreductase polypeptides. 2. The engineered ketoreductase polypeptide of claim 1 , wherein the amino acid sequence of said ketoreductase polypeptide comprises the substitution X206F/L and one or more residue differences as compared to SEQ ID NO:4 selected from: X7S; X17M; X17Q; X17R; X23V; X27L; X29G; X40R; X60I; X64V; X71P; X87L; X94A; X94P; X94S; X95M; X105G; X113I; X122A; X127R; X131S; X144V; X145L; X147I; X147L; X147Q; X150Y; X152G; X153G; X157C; X173L; X196M; X198S; X208R; X216R; X221S; X243S; X245I; X249F; X249G; and X249Y.3. The engineered ketoreductase polypeptide of claim 1 , wherein the amino acid sequence of said ketoreductase polypeptide comprises X206F/L claim 1 , and at least one or more residue differences as compared to SEQ ID NO:4 selected from: X17Q/R/M; X40R; X64V; X94P; X96L/Y; X144V; X147Q/I/L; X157C; X195A/G; X196M; and X199H.4. The engineered ketoreductase polypeptide of claim 1 , wherein the ketoreductase polypeptide is capable of converting the substrate compound (2) to the product compound (1) with at least 10 fold the activity of the reference polypeptide of SEQ ID NO:4 claim 1 , wherein the amino acid sequence comprises the substitution X206F/L claim 1 , and one or more residue differences as compared to SEQ ID NO:4 selected from: X40R; X60I; X71P; X94P; X94A; X95M; X96L; X96Y; X127R; X144V; X1451; X150Y; X152G; X153G; X157C; X195A; X195G; X196M; X198S; X199H; X216R claim 1 , X245I claim 1 , X245F; X249Y; and X249F.5. The engineered ketoreductase polypeptide of claim 1 , wherein the ketoreductase polypeptide has increased thermal stability as compared to the reference polypeptide of SEQ ID NO:4 or 32 claim 1 , wherein the amino acid sequence comprises the substitution X206F/L claim 1 , and one or more residue differences as ...

FUNCTIONALIZED CARBOXYLIC ACIDS AND ALCOHOLS BY REVERSE FATTY ACID OXIDATION

Bacteria that run the beta oxidation cycle in reverse anabolic direction are provided, along with many novel primers to start the reverse cycle, pathways to make such primers, and a large variety of products produced thereby. Methods for making desired product by using such primers in the reverse pathway are also disclosed. 1. A genetically engineered microbe comprising:a) one or more overexpressed enzymes that allow the production of an omega-functionalized CoA thioester primer selected from oxalyl-CoA, malonyl-CoA, succinyl-CoA, hydroxyacetyl-CoA, 3-hydroxypropionyl-CoA, 4-hydroxybutyryl-CoA, 2-aminoacetyl-CoA, 3-aminopropionoyl-CoA, 4-aminobutyrylCoA, isobutyryl-CoA, 3-methyl-butyryl-CoA, 2-hydroxypropionyl-CoA, 3-hydroxybutyryl-CoA, or 2-aminopropionyl-CoA;b) an overexpressed thiolase able to act on an omega-functionalized CoA thioester primer;c) an overexpressed 3-hydroxyacyl-CoA dehydrogenase able to act on an omega-functionalized 3-ketoacyl-CoA substrate;d) an overexpressed enoyl-CoA hydratase or 3-hydroxyacyl-CoA dehydratase able to act on an omega-functionalized 3-hydroxyacyl-CoA substrate;e) an overexpressed acyl-CoA dehydrogenase or trans-enoyl-CoA reductase able to act on an omega-functionalized trans-enoyl-CoA substrate; i) a thioesterase, or an acyl-CoA:acetyl-CoA transferase, or a phosphotransacylase and a carboxylate kinase;', 'ii) an alcohol-forming coenzyme-A thioester reductase;', 'iii) an aldehyde-forming CoA thioester reductase and an alcohol dehydrogenase;', 'iv) an aldehyde-forming CoA thioester reductase and an aldehyde decarbonylase;', 'v) an olefin-forming enzyme; or', 'vi) an aldehyde-forming CoA thioester reductase and a transaminase; and, 'f) an overexpressed termination enzyme able to act on an omega-functionalized CoA-thioester substrate or their beta-functionalized derivatives, wherein said termination enzyme is selected fromg) reduced expression of fermentation enzymes leading to reduced production of lactate, acetate, ethanol and ...

ENGINEERED IMINE REDUCTASES AND METHODS FOR THE REDUCTIVE AMINATION OF KETONE AND AMINE COMPOUNDS

The present disclosure provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds. 1. An engineered polypeptide having imine reductase activity , wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2 , and comprises a substitution at a position corresponding to position 197 of SEQ ID NO:2 , wherein the amino acid at position 197 has been replaced with proline or an aliphatic or a non-polar residue.2. The engineered polypeptide having imine reductase activity of claim 1 , wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2 claim 1 , wherein the amino acid at the position corresponding to position 197 of SEQ ID NO:2 has been replaced with proline.9. The process of claim 8 , in which Rand Rare linked to form a 3-membered to 10-membered ring.10. The process of claim 8 , in which the substrate compound of formula (II) is selected from methylamine claim 8 , dimethylamine claim 8 , isopropylamine claim 8 , butylamine claim 8 , isobutylaminel claim 8 , L-norvaline claim 8 , aniline claim 8 , (S)-2-aminopent-4-enoic acid claim 8 , pyrrolidine claim 8 , and hydroxypyrrolidine.11. The process of claim 8 , in which at least one of Rand Rof the compound of formula (I) is linked to at least one of Rand Rof the amine compound of formula (II) claim 8 , whereby the process for preparing the amine compound of formula (III) comprises an intramolecular reaction.12. The process of claim 8 , in which the suitable reaction conditions comprise:(a) substrate loading at about 10 g/L to 100 g/L;(b) about 0.1 g/L to about 50 g/L of the engineered polypeptide;(c) about 0.05 g/L (0.001 M) to about 2.5 g/L (0.050 M) of NAD(P)H;(d) a pH of about 6 to 10;(e) ...