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    Phenylethanol dehydrogenase capable of reducing acetophenone to R(+)-phenylethanol
    31.
    发明授权
    Phenylethanol dehydrogenase capable of reducing acetophenone to R(+)-phenylethanol 失效
    将乙炔脱氢酶还原成R（+） - 苯乙醇的苯乙醇脱氢酶

    公开(公告)号：US5200335A

    公开(公告)日：1993-04-06

    申请号：US694988

    申请日：1991-05-06

    申请人： Werner Hummel ,  Maria-Regina Kula

    发明人： Werner Hummel ,  Maria-Regina Kula

    IPC分类号： C12N9/04 ,  C12P7/02 ,  C12P7/22 ,  C12P7/26 ,  C12P41/00 ,  C12R1/225

    CPC分类号： C12N9/0006 ,  C12P41/00 ,  C12P7/02 ,  C12P7/22 ,  C12P7/26 ,  Y10S435/853

    摘要： A phenylethanol dehydrogenase capable of catalyzing the reduction of acetophenone to R(+)-phenylethanol in the presence of NADPH was isolated from Lactobacilli such as Lactobacillus kefir. The dehydrogenase is also capable of catalyzing the reduction of aromatic, alicyclic and aliphatic ketones selected from the group consisting of p-bromoacetophenone, methylcyclohexanone, acetone, methyl hexyl ketone, 4-phenyl-2-butanone, 1-phenyl-1,2-propanedione, ethyl pentyl ketone, pinacolone, propiophenone and p-chloroacetophenone. The dehydrogenase is rapidly inactivated by EDTA, but conventional inhibitors, chelators and SH-protecting reagents have only a slight effect on activity. The enzyme has a K.sub.M of 6.times.10.sup.-4 M for acetophenone. The dehydrogenase is capable of catalyzing the enzymatic reduction of carbonyl compounds to form optically active hydroxy compounds in the presence of NADPH. In such reactions, NADPH can be simultaneously regenerated in the presence of glucose 6-P and glucose-6-P dehydrogenase or isopropanol.

    摘要翻译： 从乳酸杆菌（Lactobacilli）如乳杆菌（Lactobacillus kefir）中分离能够在NADPH存在下催化苯乙酮还原为R（+） - 苯基乙醇的苯乙醇脱氢酶。 脱氢酶还能够催化选自对溴苯乙酮，甲基环己酮，丙酮，甲基己基酮，4-苯基-2-丁酮，1-苯基-1,2-苯并恶唑酮的芳族，脂环族和脂族酮的还原， 丙二酮，乙基戊基酮，频哪酮，苯丙酮和对氯苯乙酮。 脱氢酶由EDTA快速灭活，但是常规的抑制剂，螯合剂和SH-保护试剂对活性只有轻微的影响。 对于苯乙酮，该酶的KM为6×10 -4 M。 在NADPH存在下，脱氢酶能够催化羰基化合物的酶还原形成光学活性羟基化合物。 在这种反应中，NADPH可以在葡萄糖6-P和葡萄糖-6-P脱氢酶或异丙醇的存在下同时再生。

    Process for the enzymatic preparation of optically-active cyanohydrins
    32.
    发明授权
    Process for the enzymatic preparation of optically-active cyanohydrins 失效
    光活性氰基化合物的制备方法

    公开(公告)号：US5122462A

    公开(公告)日：1992-06-16

    申请号：US670437

    申请日：1991-03-18

    申请人： Peter Miethe ,  Maria-Regina Kula ,  Ingeborg M. Stuertz ,  Christian Wandrey ,  Udo Kragl

    发明人： Peter Miethe ,  Maria-Regina Kula ,  Ingeborg M. Stuertz ,  Christian Wandrey ,  Udo Kragl

    IPC分类号： C12M1/00 ,  C12M1/40 ,  C12P13/00 ,  C12P41/00

    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。

    Process for the continuous enzymatic change of water soluble .alpha.-ketocarboxylic acids into the corresponding amino acids
    34.
    发明授权
    Process for the continuous enzymatic change of water soluble .alpha.-ketocarboxylic acids into the corresponding amino acids 失效
    将水溶性α-酮羧酸连续酶促转化为相应氨基酸的方法

    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

    IPC分类号： C07C229/06 ,  C12M1/34 ,  C12P1/00 ,  C12P13/04 ,  C12P13/06 ,  C12P19/36 ,  C12P13/08

    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。

    Method for isolating an enzyme from a fermentation broth
    39.
    发明授权
    Method for isolating an enzyme from a fermentation broth 失效
    从发酵液中分离酶的方法

    公开(公告)号：US5330905A

    公开(公告)日：1994-07-19

    申请号：US750305

    申请日：1991-08-27

    申请人： Maria-Regina Kula ,  Martin-Roger Grote

    发明人： Maria-Regina Kula ,  Martin-Roger Grote

    IPC分类号： C12M1/40 ,  C12N9/00 ,  C12N9/28 ,  C12N9/32 ,  C12N9/50

    CPC分类号： C12M21/18 ,  C12M23/06 ,  C12M47/10 ,  C12N9/00

    摘要： To obtain enzymes from an enzyme-containing suspension, such as fermentation broth, the suspension is placed, together with particulate material capable of binding enzyme, such as ion exchange resin, in a vertical container. An ascending stream of inert gas, such as dry nitrogen, is passed through the resulting composition at a rate sufficient for fluidization. The resulting composition is mixed and evaporated until an adequate amount of enzyme is bound to the particulate material. Concentration of enzyme in the suspension is directly proportional to the amount of enzyme bound to particulate material. The residual suspension is forced out of the container by a pulse of compressed gas, and the bound enzyme is eluted. To speed up the process, the stream of gas and/or the composition are heated to enzyme-tolerated temperatures. The resulting composition can be further subjected to a washing step after removal of the residual suspension and prior to elution of the enzyme. The washing liquid can be passed through another fixed bed column containing particulate material capable of binding enzyme. Elution of the enzyme can be carried out as a stepwise process.

    摘要翻译： 为了从含酶的悬浮液如发酵液中获得酶，将悬浮液与能够结合酶的颗粒物质一起放置在立式容器中，例如离子交换树脂。 惰性气体如干燥氮气的上升流以足以流化的速率通过所得组合物。 将所得组合物混合并蒸发，直到足够量的酶与颗粒物质结合。 悬浮液中酶的浓度与与颗粒物质结合的酶的量成正比。 残留的悬浮液被压缩气体的脉冲压出容器，并且结合的酶被洗脱。 为了加速该过程，将气流和/或组合物加热至耐受酶的温度。 所得组合物可以在去除残留悬浮液之后并在酶洗脱之前进一步进行洗涤步骤。 洗涤液可以通过含有能结合酶的颗粒材料的另一固定床柱。 酶的洗脱可以作为逐步方法进行。

    Method of using N-acetyl-2,3-Didehydroleucine acylase for the preparation of D- or L-tryptophyl glycine, D- or L-tryptophyl-D-methionine or L-tryptophyl-D-cysteine
    40.
    发明授权
    Method of using N-acetyl-2,3-Didehydroleucine acylase for the preparation of D- or L-tryptophyl glycine, D- or L-tryptophyl-D-methionine or L-tryptophyl-D-cysteine 失效
    使用N-乙酰基-2,3-二脱氢荧光素酰基转移酶制备D-或L-色氨酸甘氨酸，D-或L-色氨酰-D-甲硫氨酸或L-色氨酰-D-半胱氨酸的方法

    公开(公告)号：US5212069A

    公开(公告)日：1993-05-18

    申请号：US870817

    申请日：1992-04-20

    申请人： Maria-Regina Kula ,  Matthias Kittelmann

    发明人： Maria-Regina Kula ,  Matthias Kittelmann

    IPC分类号： C12N9/80 ,  C12P13/06

    CPC分类号： C12P13/06 ,  C12N9/80

    摘要： A novel N-Acetyl-2,3-didehydroaminoacid-acylase is obtained by cultivating Zoogloea ramigera DSM 4306. The new enzyme can be used in a coupled enzyme system with an L-Leucinedehydrogenase for the enzymatic conversion of N-Acetyl-2,3-didehydroleucine to L-Leucine, D- or L-tryptophylglycine to D- or L-tryptophaneamide and glycine, as well as other tryptophanedipeptides to tryptophaneamides and free amino acids.

    摘要翻译： 通过培养Zoogloea ramigera DSM 4306获得新的N-乙酰基-2,3-二脱氢氨基酸酰基转移酶。该新酶可用于具有L-亮氨酸氢化酶的偶联酶系统中用于N-乙酰基-2,3 D-脱水亮氨酸向L-亮氨酸，D-或L-色氨酰氨基甘氨酸转化为D-或L-色氨酰胺和甘氨酸，以及其他色氨酸肽与色氨酰胺和游离氨基酸。