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1.发明授权
公开(公告)号：US4326031A
公开(公告)日：1982-04-20
申请号：US172444
申请日：1980-07-25
申请人： Christian Wandrey , Rolf Wichmann , Wolfgang Leuchtenberger , Maria-Regina Kula , Andreas Buckmann
发明人： Christian Wandrey , Rolf Wichmann , Wolfgang Leuchtenberger , Maria-Regina Kula , Andreas Buckmann
摘要： Water soluble .alpha.-ketocarboxylic acids are continuously converted in a membrane reactor into the corresponding .alpha.-hydrocarboxylic acids. The conversion takes place in the presence of a substrate specific dehydrogenase and of a nicotinamide-adenine-dinucleotide (NAD.sup.+ /NADH) enlarged in molecular weight through linkage to a water soluble polymer as coenzyme. Simultaneously NADH is regenerated continuously from NAD.sup.+ in presence of a formate dehydrogenase and from formate ion. The membrane must have a mean pore diameter of 1 to 3 nm. As coenzyme there is employed 0.1 to 10 mmol/l of NAD.sup.+ /NADH present bound to a water soluble polymer having an average molecular weight between 500 and 50,000. There is continuously supplied to the reactor a substrate stream which contains 50 to 100% of the maximum amount soluble, but not over 2,000 mmol/l, of the reacting .alpha.-ketocarboxylic acid in the form of a water soluble salt and 100 to 6,000 mmol/l of a formate. There is maintained over the membrane a differential pressure of 0.1 to 15 bar. There is continuously drawn off behind the membrane a filtrate stream containing the .alpha.-hydroxy acid formed.
摘要翻译： 水溶性α-酮羧酸在膜反应器中连续转化成相应的α-羟基羧酸。 通过与作为辅酶的水溶性聚合物连接，在底物特异性脱氢酶和分子量增大的烟酰胺 - 腺嘌呤二核苷酸（NAD + / NADH）的存在下进行转化。 同时NADH在甲酸脱氢酶和甲酸根离子存在下从NAD +连续再生。 膜的平均孔径必须为1〜3nm。 作为辅酶，使用0.1〜10mmol / l的NAD + / NADH，其与平均分子量在500〜50,000的水溶性聚合物结合。 向反应器中连续供应底物物流，其含有50〜100％最大量可溶性但不超过2,000mmol / l的水溶性盐形式的α-酮羧酸反应物和100至6,000mmol / l的甲酸盐。 在膜上保持0.1至15巴的压差。 在膜的后面连续地抽出含有形成的α-羟基酸的滤液流。
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2.发明授权
公开(公告)号：US4782020A
公开(公告)日：1988-11-01
申请号：US83882
申请日：1987-08-10
CPC分类号： C12P13/06 , C12P13/04 , C12P13/12 , Y10S435/813
摘要： .alpha.-Hydroxycarboxylic acids are continuously converted into the corresponding optically active .alpha.- aminocarboxylic acids. The conversion is carried out in a membrane reactor in the presence of nicotinamide-adenine dinucleotide increased in molecular weight by bonding to a water soluble high molecular weight material, a dehydrogenase specific for the .alpha.-hydroxycarboxylic acid, a dehydrogenase specific for the corresponding .alpha.-amino-carboxylic acid and ammonium ions. There is continuously supplied to the membrane reactor an aqueous solution of the .alpha.-hydroxycarboxylic acid to be reacted, a substantially lesser amount of the corresponding .alpha.-ketocarboxy lic acid, and an amount of ammonium ion at least equivalent to the .alpha.-hydroxycarboxylic acid to be reacted. There is maintained over the membrane a difference in pressure 1 and 15 bar. Behind the membrane, there is continuously drawn off a filtrate stream containing the .alpha.-aminocarboxylic acid formed.
摘要翻译： α-羟基羧酸连续转化为相应的光学活性α-氨基羧酸。 在通过结合到水溶性高分子量材料的烟酰胺 - 腺嘌呤二核苷酸增加分子量的存在下，在膜反应器中进行转化，对于α-羟基羧酸是特异性的脱氢酶，对于相应的α- 氨基羧酸和铵离子。 向膜反应器中连续供应待反应的α-羟基羧酸的水溶液，相当少量的相应的α-酮羧酸的丙酸，以及一定量的至少等同于α-羟基羧酸的铵离子至 被反应。 在膜上保持压力1和15巴的差异。 在膜的后面，连续地抽出含有形成的α-氨基羧酸的滤液流。
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3.发明授权
公开(公告)号：US4304858A
公开(公告)日：1981-12-08
申请号：US172446
申请日：1980-07-25
申请人： Christian Wandrey , Rolf Wichmann , Wolfgang Leuchtenberger , Maria-Regina Kula , Andreas Buckmann
发明人： Christian Wandrey , Rolf Wichmann , Wolfgang Leuchtenberger , Maria-Regina Kula , Andreas Buckmann
CPC分类号： C12P13/06 , C12P1/00 , C12P13/04 , C12P19/36 , Y10S435/813
摘要： Water soluble .alpha.-ketocarboxylic acids are continuously converted in a membrane reactor into the corresponding aminoacids. The conversion takes place in the presence of a substrate specific dehydrogenase, of ammonium ions and of a nicotinamide-adenine-dinucleotide (NAD.sup.+ /NADH) enlarged in molecular weight through linkage to a water soluble polymer as coenzyme. Simultaneously NADH is regenerated continuously from NAD.sup.+ in presence of a formate dehydrogenase and from formate ion. The membrane must have a mean pore diameter of 1 to 3 nm. As coenzyme there is employed 0.1 to 10 mmol/l of NAD.sup.+ /NADH present bound to a polyoxyethylene having an average molecular weight between 500 and 50,000. There is continuously supplied to the reactor a substrate stream which contains 50 to 100% of the maximum amount soluble, but not over 2,000 mmol/l, of the reacting .alpha.-ketocarboxylic acid in the form of a water soluble salt, an ammonium ion in an amount about equimolar to the amount of substrate and 100 to 6,000 mmol/l of a formate. There is maintained over the membrane a differential pressure of 0.1 to 15 bar. There is continuously drawn off behind the membrane a filtrate stream containing the aminoacid formed.
摘要翻译： 水溶性α-酮羧酸在膜反应器中连续转化成相应的氨基酸。 通过与作为辅酶的水溶性聚合物连接，在底物特异性脱氢酶，铵离子和烟酰胺 - 腺嘌呤二核苷酸（NAD + / NADH）存在下进行转化，分子量增加。 同时NADH在甲酸脱氢酶和甲酸根离子存在下从NAD +连续再生。 膜的平均孔径必须为1〜3nm。 作为辅酶，使用0.1〜10mmol / l的NAD + / NADH，其与平均分子量在500〜50,000的聚氧乙烯结合。 向反应器中连续供应底物流，其含有50〜100％最大量可溶性但不超过2,000mmol / l的水溶性盐，铵离子形式的α-酮羧酸 相当于底物量的等摩尔量和100-6,000mmol / l甲酸盐的量。 在膜上保持0.1至15巴的压差。 在膜的后面连续抽出含有形成的氨基酸的滤液流。
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公开(公告)号：US5369016A
公开(公告)日：1994-11-29
申请号：US005819
申请日：1993-01-19
摘要： A peptide amidase isolated from the flavedo of citrus fruits, preferably oranges, which is capable of catalyzing the selective hydrolytic elimination of the free amino group on the C-terminal end of peptide amides but which does not cleave peptide bonds. The enzyme accepts D-amino acid residues in the C-terminal position, although the hydrolysis rate is much slower than with L-amino acid residues. The enzyme is weakly inhibited by serine protease inhibitors; has an optimal pH of 7.5.+-.1.5, an optimum temperature of 30.degree. C. at pH 7.5 and has an isoelectric point of pH 9.5. The peptide amidase is stable at pH 6.0-9.0. The molecular weight of the purified enzyme is 32,000.+-.3000 daltons. A peptide amidase according to the present invention is particularly useful in the production of peptides by continuous enzymatic reaction of N-protected amino acid or peptide alkyl esters with amides of amino acids. In the continuous reaction, the synthesized peptide amide is hydrolyzed by the peptide amidase and separated by anion exchange from the amide of the amino acid which can be recycled.
摘要翻译： 从柑橘类水果（优选橙子）中分离的肽酰胺酶，其能够催化肽酰胺的C末端上的游离氨基的选择性水解消除，但不切割肽键。 该酶在C-末端位置接受D-氨基酸残基，尽管水解速度比L-氨基酸残基慢得多。 酶被丝氨酸蛋白酶抑制剂弱抑制; 最佳pH为7.5 +/- 1.5，最适温度为30℃，pH 7.5，等电点pH值为9.5。 肽酰胺酶在pH 6.0-9.0下是稳定的。 纯化酶的分子量为32,000 +/- 3000道尔顿。 根据本发明的肽酰胺酶特别可用于通过N-保护的氨基酸或肽烷基酯与氨基酸酰胺的连续酶反应来生产肽。 在连续反应中，合成的肽酰胺被肽酰胺酶水解，并通过阴离子交换从可再循环的氨基酸的酰胺中分离。
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公开(公告)号：US5190875A
公开(公告)日：1993-03-02
申请号：US694981
申请日：1991-05-06
摘要： A peptide amidase isolated from the flavedo of citrus fruits, preferably oranges, which is capable of catalyzing the selective hydrolytic elimination of the free amino group on the C-terminal end of peptide amides but which does not cleave peptide bonds. The enzyme accepts D-amino acid residues in the C-terminal position, although the hydrolysis rate is much slower than with L-amino acid residues. The enzyme is weakly inhibited by serine protease inhibitors; has an optimal pH of 7.5.+-.1.5, an optimum temperature of 30.degree. C. at pH 7.5 and has an isoelectric point of pH 9.5. The peptide amidase is stable at pH 6.0-9.0. The molecular weight of the purified enzyme is 23,000 +/- 3000 daltons. A peptide amidase according to the present invention is particular useful in the production of peptides by continuous enzymatic reaction of N-protected amino acid or peptide alkyl esters with amides of amino acids. In the continuous reaction, the synthesized peptide amide is hydrolyzed by the peptide amidase and separated by anion exchange from the amide of the amino acid which can be recycled.
摘要翻译： 从柑橘类水果（优选橙子）中分离的肽酰胺酶，其能够催化肽酰胺的C末端上的游离氨基的选择性水解消除，但不切割肽键。 该酶在C-末端位置接受D-氨基酸残基，尽管水解速度比L-氨基酸残基慢得多。 酶被丝氨酸蛋白酶抑制剂弱抑制; 最佳pH为7.5 +/- 1.5，最适温度为30℃，pH 7.5，等电点pH值为9.5。 肽酰胺酶在pH 6.0-9.0下是稳定的。 纯化酶的分子量为23000 +/- 3000道尔顿。 根据本发明的肽酰胺酶特别可用于通过N-保护的氨基酸或肽烷基酯与氨基酸的酰胺的连续酶反应来生产肽。 在连续反应中，合成的肽酰胺被肽酰胺酶水解，并通过阴离子交换从可再循环的氨基酸的酰胺中分离。
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公开(公告)号：US5122462A
公开(公告)日：1992-06-16
申请号：US670437
申请日：1991-03-18
CPC分类号： C12P13/004 , C12M21/18 , C09K2219/15
摘要： The preparation of optically-active cyanohydrins from the corresponding oxo compounds is disclosed. The reaction of the oxo compounds with hydrocyanic acid is carried out in an organic solvent in the presence of (R)- or (S)-oxynitrilase (4.1.2.10) and (4.1.2.11), respectively, being solubilized in a lyotropic liquid crystal. Compounds which upon hydrolysis produce increased pH values are excluded as surface active agents. Preferably, surface active agents, the organic solvent and an aqueous buffer solution with a pH of 3 to 6, are mixed together to obtain a liquid crystal/organic solvent two-phase system. The liquid crystal is preferably fixed on a porous support, in particular a glass support. The reaction is carried out in a flow-through reactor which contains the liquid crystal in the abovementioned form or in thin layers adjacent to narrow flow channels, the borders of which are liquid permeable and through which the substrate-containing solvent is passed.
摘要翻译： 公开了从相应的氧代化合物制备光学活性的氰醇。 氧代化合物与氢氰酸的反应在有机溶剂中分别在（R） - 或（S） - 硝基腈酶（4.1.2.10）和（4.1.2.11）的存在下进行，溶解在溶致液体 水晶。 在水解产生增加的pH值的化合物被排除作为表面活性剂。 优选将表面活性剂，有机溶剂和pH为3〜6的缓冲水溶液混合在一起，得到液晶/有机溶剂两相体系。 液晶优选固定在多孔载体上，特别是玻璃载体上。 该反应在含有上述形式的液晶的流通反应器中或邻近窄流道的薄层中进行，该流动通道的边界是液体可渗透的并且含底物的溶剂通过该反应器。
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7.发明授权Process for the fermentative preparation of L-amino acids from .alpha.-keto carboxylic acids 失效来自（ALPHA） - 羧基羧酸的L-氨基酸的发酵制备方法
公开(公告)号：US5053328A
公开(公告)日：1991-10-01
申请号：US908777
申请日：1986-09-18
CPC分类号： C12P13/06 , C12P13/04 , Y10S435/832 , Y10S435/833 , Y10S435/834 , Y10S435/835 , Y10S435/836 , Y10S435/837 , Y10S435/838 , Y10S435/839
摘要： The preparation of an L-amino acid, particularly valine, leucine, isoleucine, alanine and phenylalanine, from the corresponding .alpha.-keto carboxylic acid by bacterial fermentation in the presence of ammonium ions is carried out with the aid of thermophilic Bacillus strains at temperatures above 45.degree. C., in particular above 60.degree. C. Bacillus strains DSM 406, 452, 461, 42, 463, 465 and 466 are particularly suitable for this purpose. The greater solubility of the amino acid at the elevated fermentation temperature permits the separation out of the amino acid from the reaction mixture simply by cooling, whereafter the depleted reaction mixture can be pumped back into the fermenter. Especially favorable yields are achieved by supplying oxygen to the fermenter to an amount of less than about 20% dissolved oxygen.
摘要翻译： 在铵离子存在下通过细菌发酵从相应的α-酮羧酸制备L-氨基酸，特别是缬氨酸，亮氨酸，异亮氨酸，丙氨酸和苯丙氨酸，借助于嗜热芽孢杆菌菌株，在温度高于 45℃，特别是高于60℃。芽孢杆菌菌株DSM 406,452,461,42,463,465和466特别适用于此目的。 氨基酸在升高的发酵温度下的更大的溶解度允许简单地通过冷却从反应混合物中分离出氨基酸，然后将贫化的反应混合物泵送回发酵罐。 通过向发酵罐供应氧气至小于约20％的溶解氧的量来实现特别有利的产率。
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8.发明授权Method for the fermentative production of L-amino acids from .alpha.-keto acids 失效从（ALPHA） - 酮酸生成L-氨基酸的方法
公开(公告)号：US5200326A
公开(公告)日：1993-04-06
申请号：US44678
申请日：1987-05-01
申请人： Christian Wandrey , Rolf Wichmann , Ulrich Groeger , Manfred Kircher , Wolfgang Leuchtenberger , Eberhard Breuker
发明人： Christian Wandrey , Rolf Wichmann , Ulrich Groeger , Manfred Kircher , Wolfgang Leuchtenberger , Eberhard Breuker
CPC分类号： C12P13/06 , C12P13/04 , Y10S435/84 , Y10S435/843
摘要： The invention is directed to a method of producing L-amino acids from .alpha.-ketocarboxylic acids by means of continuous fermentation with the aid of glutamate producing bacteria with biomass retention in which the cells are separated by microfiltration and/or centrifugal separators.
摘要翻译： 本发明涉及一种通过在具有生物质保留的谷氨酸生产细菌的帮助下通过连续发酵从α-酮羧酸生产L-氨基酸的方法，其中通过微量过滤和/或离心分离器分离细胞。
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公开(公告)号：US06303351B1
公开(公告)日：2001-10-16
申请号：US08208123
申请日：1994-03-08
IPC分类号： C12N744
CPC分类号： C12P7/48 , C07C59/265
摘要： A process for producing citric acid/citrate for use in producing citric acid, comprising the steps of continuously feeding to a fermenter containing a yeast capable of transforming glucose to citric acid by fermentation, in the presence of a nutrient medium and oxygen, sugar, especially glucose as a C source and at least one ammoniun compound as an N source fermenting the sugar to citric acid in said fermenter at a temperature, a pH and with a residence time sufficient to transform at least a major part of the sugar continuously fed to said fermenter to citric acid continuously withdrawing fermentation product from said fermenter recovering citric acid/citrate from said fermentation product controlling a C/N ratio fed to said fermenter to correspond to a molar ratio of glucose to ammonium compounds of 12 to 22 during fermentation of the glucose to citrate.
摘要翻译： 一种生产用于生产柠檬酸的柠檬酸/柠檬酸盐的方法，包括以下步骤：在营养培养基和氧气，糖，特别是存在下，连续喂食含有能够通过发酵将葡萄糖转化为柠檬酸的酵母的发酵罐 葡萄糖作为C源和至少一种作为N源的铵化合物，在所述发酵罐中，在温度，pH和停留时间内将糖发酵成柠檬酸，并且具有足以将至少大部分连续供给至所述 发酵罐至柠檬酸连续地从所述发酵罐抽出发酵产物，从所述发酵产物回收柠檬酸/柠檬酸，控制进料至所述发酵罐的C / N比值，以对应葡萄糖发酵期间葡萄糖与铵化合物的摩尔比为12至22 到柠檬酸盐。
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10.发明授权
公开(公告)号：US5795750A
公开(公告)日：1998-08-18
申请号：US826881
申请日：1997-04-07
申请人： Wolfgang Kruse , Udo Kragl , Christian Wandrey
发明人： Wolfgang Kruse , Udo Kragl , Christian Wandrey
IPC分类号： C12M1/40 , C12P1/00 , C12P7/04 , C12P7/22 , C12P7/26 , C12P13/00 , C12M1/12 , C12N11/00 , C12P7/64
CPC分类号： C12P1/00 , C12M21/18 , C12M29/04 , C12M29/18 , C12P7/04 , C12P7/26 , Y02P20/582 , Y10S435/813 , Y10S435/817
摘要： In a process for the continuous enzyme-catalyticzed enzymatic production of hydrophobic products in an aqueous solution of a reaction mixture, the product is extracted from the product containing reaction mixture by way of a product-permeable microporous membrane into an organic solvent and the product-depleted reaction mixture is recycled.
摘要翻译： 在反应混合物的水溶液中连续酶催化生产疏水产物的方法中，通过产物渗透微孔膜将产物从含产物的反应混合物中提取到有机溶剂中， 贫化的反应混合物被回收。
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