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    NADH OXIDASE MUTANT HAVING IMPROVED STABILITY AND USE THEREOF
    3.
    发明申请
    NADH OXIDASE MUTANT HAVING IMPROVED STABILITY AND USE THEREOF 有权
    具有改善稳定性的NADH氧化物突变体及其用途

    公开(公告)号：US20130030164A1

    公开(公告)日：2013-01-31

    申请号：US13574458

    申请日：2011-01-19

    申请人： Shinichi Yoshida ,  Akira Iwasaki ,  Motohisa Washida ,  Tozo Nishiyama ,  Daisuke Moriyama ,  Naoaki Taoka

    发明人： Shinichi Yoshida ,  Akira Iwasaki ,  Motohisa Washida ,  Tozo Nishiyama ,  Daisuke Moriyama ,  Naoaki Taoka

    IPC分类号： C12N9/02 ,  C12N15/63 ,  C07H21/02 ,  C12N1/19 ,  C12N5/10 ,  C12P19/36 ,  C12N15/53 ,  C12N1/21

    CPC分类号： C12N9/0036 ,  C12N9/0065 ,  C12P7/02 ,  C12P7/18 ,  C12P7/42 ,  C12P13/04 ,  C12P19/36 ,  C12Y106/03 ,  C12Y106/03001 ,  C12Y111/01001

    摘要： Water-forming NADH oxidase derived from Streptococcus mutans should be further improved in terms of stability for practical use in industrial production. An object of the present invention is to provide an enzyme that is obtained through modification of a water-forming NADH oxidase, which is useful as an NAD+ regeneration system for stereoselective oxidation catalyzed by an oxidoreductase, by protein engineering techniques so that the enzyme can withstand long-term use without exhibiting a reduction of its activity for the regeneration of NAD+, that is, an enzyme having improved stability, and to provide a method for efficiently producing a useful substance such as an optically active alcohol or amino acid. The present invention relates to an enzyme modification method that can improve the stability of water-forming NADH oxidase derived from Streptococcus mutans by appropriately introducing mutation.

    摘要翻译： 来自变形链球菌的成水NADH氧化酶在工业生产实际使用的稳定性方面应进一步提高。 本发明的目的是提供通过蛋白质工程技术，通过改性水合NADH氧化酶获得的酶，其可用作由氧化还原酶催化的用于立体选择性氧化的NAD +再生系统，使得酶能够承受 长期使用而不显示其NAD +再生活性的活性降低，即稳定性提高的酶，以及提供有效生产光学活性醇或氨基酸等有用物质的方法。 本发明涉及通过适当地引入突变来提高来自变形链球菌的水形成NADH氧化酶的稳定性的酶修饰方法。

    Method for production of erythro-or threo-2-amino-3-hydroxypropionic acid ester, novel carbonyl reductase, gene for the reductase, vector, transformant, and method for production of optically active alcohol using those
    4.
    发明授权
    Method for production of erythro-or threo-2-amino-3-hydroxypropionic acid ester, novel carbonyl reductase, gene for the reductase, vector, transformant, and method for production of optically active alcohol using those 有权
    用于生产红血球或苏氨酸-2-氨基-3-羟基丙酸酯的方法，新的羰基还原酶，还原酶的基因，载体，转化体和使用这些的生产光学活性醇的方法

    公开(公告)号：US08304216B2

    公开(公告)日：2012-11-06

    申请号：US12225683

    申请日：2007-03-29

    申请人： Tozo Nishiyama ,  Yoshihiko Yasohara

    发明人： Tozo Nishiyama ,  Yoshihiko Yasohara

    IPC分类号： C12P7/02 ,  C12P7/62 ,  C07H21/04

    CPC分类号： C12P13/04 ,  C12N9/0006

    摘要： The present invention has its object to provide a method of producing an erythro- or threo-2-amino-3-hydroxypropionic acid ester, and so forth. The present invention relates to: a method of asymmetrically reducing an N-2-amino-3-oxopropionic acid ester by allowing cells of a microorganism to act thereon; a polypeptide having an activity of asymmetrically reducing a carbonyl compound to give an optically active alcohol, which is isolated from a microorganism belonging to genus Brevundimonas; a DNA coding for the polypeptide; and a transformant producing the polypeptide. The invention also relates to a method of producing an optically active alcohol by reducing a carbonyl compound with the help of the polypeptide or the transformant.

    摘要翻译： 本发明的目的是提供一种制备赤式或苏氨酸-2-氨基-3-羟基丙酸酯的方法等。 本发明涉及：通过使微生物的细胞在其上作用而不对称地还原N-2-氨基-3-氧代丙酸酯的方法; 具有不对称还原羰基化合物的活性的多肽，得到从Brevundimonas属的微生物分离的光学活性醇; 编码该多肽的DNA; 和产生多肽的转化体。 本发明还涉及通过在多肽或转化体的帮助下还原羰基化合物来制备光学活性醇的方法。

    METHOD FOR PRODUCING L-AMINO ACID
    5.
    发明申请
    METHOD FOR PRODUCING L-AMINO ACID 有权
    生产L-氨基酸的方法

    公开(公告)号：US20110229940A1

    公开(公告)日：2011-09-22

    申请号：US13126534

    申请日：2009-10-28

    申请人： Masutoshi Nojiri ,  Tozo Nishiyama ,  Naoaki Taoka

    发明人： Masutoshi Nojiri ,  Tozo Nishiyama ,  Naoaki Taoka

    IPC分类号： C12P13/04 ,  C07C229/36 ,  C07C229/30

    CPC分类号： C12P13/04 ,  C07C227/30 ,  C07C233/47 ,  C12N9/104 ,  C12N9/90 ,  C12P13/222 ,  C12P41/007 ,  Y02P20/52 ,  C07C229/30 ,  C07C229/32 ,  C07C229/36

    摘要： The present invention relates to a method for producing an L-amino acid by reacting an enantiomeric mixture of an N-succinyl amino acid with L-succinylase in the presence of N-acylamino acid racemase to specifically hydrolyze the L-form. In particular, the present invention relates to a method for producing an L-amino acid in high yield by using an N-succinyl amino acid whose dissolved concentration is particularly low as a raw material to perform a reaction while precipitating the produced L-amino acid out of the reaction system. The present invention enables efficient production of an L-amino acid having high optical purity, particularly an L-amino acid useful as a raw material for products such as pharmaceutical products and agricultural chemicals.

    摘要翻译： 本发明涉及通过在N-酰基氨基酸消旋酶存在下使N-琥珀酰氨基酸与L-琥珀酰基酶的对映异构体混合物反应以特异性水解L型而生产L-氨基酸的方法。 特别地，本发明涉及通过使用溶解浓度特别低的N-琥珀酰氨基酸作为原料进行反应，同时使生成的L-氨基酸沉淀而以高产率生产L-氨基酸的方法 出了反应体系。 本发明能够有效地生产具有高光学纯度的L-氨基酸，特别是可用作药物产品和农药等产品的原料的L-氨基酸。

    Novel carbonyl reductase, gene thereof and method of using the same
    7.
    发明申请
    Novel carbonyl reductase, gene thereof and method of using the same 有权
    新型羰基还原酶，其基因及其使用方法

    公开(公告)号：US20070178565A1

    公开(公告)日：2007-08-02

    申请号：US11730549

    申请日：2007-04-02

    申请人： Noriyuki Kizaki ,  Tozo Nishiyama ,  Yoshihiko Yasohara

    发明人： Noriyuki Kizaki ,  Tozo Nishiyama ,  Yoshihiko Yasohara

    IPC分类号： C12P17/04 ,  C07H21/04 ,  C12P21/06 ,  C12N9/06 ,  C12N1/21 ,  C12N15/74

    CPC分类号： C12P17/10 ,  C12N9/0006 ,  C12P7/02 ,  C12P7/22

    摘要： The present invention provides a novel polypeptide efficiently forming (R)-N-benzyl-3-pyrrolidinol, a polynucleotide coding for said polypeptide, and use of the same. The present invention relates to a polypeptide having the following physical and chemical properties (1) to (4): (1) activity: acting on N-benzyl-3-pyrrolidinone with NADH or NADPH as a coenzyme, to form (R)-N-benzyl-3-pyrrolidinol; (2) optimum pH for activity: 5.5 to 6.0; (3) optimum temperature for activity: 50° C. to 55° C.; (4) molecular weight: about 55,000 as determined by gel filtration analysis, about 28,000 as determined by SDS polyacrylamide gel electrophoresis analysis. The present invention also relates to a polypeptide comprising the amino acid sequence shown under SEQ ID NO:1 in the sequence listing, a polynucleotide coding for said polypeptide, and a transformant producing said polypeptide at high levels.

    摘要翻译： 本发明提供了有效形成（R）-N-苄基-3-吡咯烷醇的新型多肽，编码所述多肽的多核苷酸及其用途。 本发明涉及具有以下物理和化学性质（1）至（4）的多肽：（1）活性：以NADH或NADPH作为辅酶作用于N-苄基-3-吡咯烷酮，形成（R） - N-苄基-3-吡咯烷醇; （2）活性最佳pH：5.5〜6.0; （3）活性最佳温度：50〜55℃。 （4）通过凝胶过滤分析确定的分子量：约55,000，通过SDS聚丙烯酰胺凝胶电泳分析确定约28,000。 本发明还涉及包含序列表中SEQ ID NO：1所示氨基酸序列的多肽，编码所述多肽的多核苷酸，以及以高水平产生所述多肽的转化体。

    Carbonyl reductase, gene thereof and method of using the same
    8.
    发明授权
    Carbonyl reductase, gene thereof and method of using the same 有权
    羰基还原酶，其基因及其使用方法

    公开(公告)号：US07220564B2

    公开(公告)日：2007-05-22

    申请号：US10528000

    申请日：2003-09-19

    申请人： Noriyuki Kizaki ,  Tozo Nishiyama ,  Yoshihiko Yasohara

    发明人： Noriyuki Kizaki ,  Tozo Nishiyama ,  Yoshihiko Yasohara

    IPC分类号： C12N9/02 ,  C12N9/52 ,  C12N1/20 ,  C12P7/22 ,  C12P21/06 ,  C07H21/02

    CPC分类号： C12P17/10 ,  C12N9/0006 ,  C12P7/02 ,  C12P7/22

    摘要： The present invention provides a novel polypeptide efficiently forming (R)-N-benzyl-3-pyrrolidinol, a polynucleotide coding for said polypeptide, and use of the same.The present invention relates to a polypeptide having the following physical and chemical properties (1) to (4): (1) activity: acting on N-benzyl-3-pyrrolidinone with NADH or NADPH as a coenzyme, to form (R)-N-benzyl-3-pyrrolidinol; (2) optimum pH for activity: 5.5 to 6.0; (3) optimum temperature for activity: 50° C. to 55° C.; (4) molecular weight: about 55,000 as determined by gel filtration analysis, about 28,000 as determined by SDS polyacrylamide gel electrophoresis analysis. The present invention also relates to a polypeptide comprising the amino acid sequence shown under SEQ ID NO:1 in the sequence listing, a polynucleotide coding for said polypeptide, and a transformant producing said polypeptide at high levels.

    摘要翻译： 本发明提供了有效形成（R）-N-苄基-3-吡咯烷醇的新型多肽，编码所述多肽的多核苷酸及其用途。 本发明涉及具有以下物理和化学性质（1）至（4）的多肽：（1）活性：以NADH或NADPH作为辅酶作用于N-苄基-3-吡咯烷酮，形成（R） - N-苄基-3-吡咯烷醇; （2）活性最佳pH：5.5〜6.0; （3）活性最佳温度：50〜55℃。 （4）通过凝胶过滤分析确定的分子量：约55,000，通过SDS聚丙烯酰胺凝胶电泳分析确定约28,000。 本发明还涉及包含序列表中SEQ ID NO：1所示氨基酸序列的多肽，编码所述多肽的多核苷酸，以及以高水平产生所述多肽的转化体。

    Method for producing L-amino acid
    9.
    发明授权
    Method for producing L-amino acid 有权
    生产L-氨基酸的方法

    公开(公告)号：US09464306B2

    公开(公告)日：2016-10-11

    申请号：US13126534

    申请日：2009-10-28

    申请人： Masutoshi Nojiri ,  Tozo Nishiyama ,  Naoaki Taoka

    发明人： Masutoshi Nojiri ,  Tozo Nishiyama ,  Naoaki Taoka

    IPC分类号： C12P13/04 ,  C12P13/00 ,  C12N9/10 ,  C12N9/90 ,  C07C227/30 ,  C07C233/47 ,  C12P13/22 ,  C12P41/00

    CPC分类号： C12P13/04 ,  C07C227/30 ,  C07C233/47 ,  C12N9/104 ,  C12N9/90 ,  C12P13/222 ,  C12P41/007 ,  Y02P20/52 ,  C07C229/30 ,  C07C229/32 ,  C07C229/36

    摘要： The present invention relates to a method for producing an L-amino acid by reacting an enantiomeric mixture of an N-succinyl amino acid with L-succinylase in the presence of N-acylamino acid racemase to specifically hydrolyze the L-form. In particular, the present invention relates to a method for producing an L-amino acid in high yield by using an N-succinyl amino acid whose dissolved concentration is particularly low as a raw material to perform a reaction while precipitating the produced L-amino acid out of the reaction system. The present invention enables efficient production of an L-amino acid having high optical purity, particularly an L-amino acid useful as a raw material for products such as pharmaceutical products and agricultural chemicals.

    摘要翻译： 本发明涉及通过在N-酰基氨基酸消旋酶存在下使N-琥珀酰氨基酸与L-琥珀酰基酶的对映异构体混合物反应以特异性水解L型而生产L-氨基酸的方法。 特别地，本发明涉及通过使用溶解浓度特别低的N-琥珀酰氨基酸作为原料进行反应，同时使生成的L-氨基酸沉淀而以高产率生产L-氨基酸的方法 出了反应体系。 本发明能够有效地生产具有高光学纯度的L-氨基酸，特别是可用作药物产品和农药等产品的原料的L-氨基酸。

    Gene suitable for alcohol dehydrogenase, vector and transformant
    10.
    发明授权
    Gene suitable for alcohol dehydrogenase, vector and transformant 有权
    基因适用于醇脱氢酶，载体和转化体

    公开(公告)号：US08129163B2

    公开(公告)日：2012-03-06

    申请号：US12516388

    申请日：2007-11-27

    申请人： Shigeru Kawano ,  Takeru Ishige ,  Keita Iguchi ,  Tozo Nishiyama ,  Yoshihiko Yasohara

    发明人： Shigeru Kawano ,  Takeru Ishige ,  Keita Iguchi ,  Tozo Nishiyama ,  Yoshihiko Yasohara

    IPC分类号： C12N9/00 ,  C12N9/02 ,  C12N1/20 ,  C12N15/00 ,  C07H21/04

    CPC分类号： C12P7/22 ,  C12N9/0006 ,  C12P7/04 ,  Y02E50/17

    摘要： An object of the present invention is to provide a novel alcohol dehydrogenase, a gene for the alcohol dehydrogenase, a vector including the gene, a transformant transformed with the vector, and a method for producing an optically active alcohol by utilizing them. A feature of the present invention directs to a novel polypeptide isolated from Candida maltosa, a DNA coding for the polypeptide, and a transformant producing the polypeptide. Another feature of the present invention directs to a method for producing an optically-active alcohol by reducing a carbonyl compound with the polypeptide or the transformant.

    摘要翻译： 本发明的目的是提供一种新的醇脱氢酶，用于醇脱氢酶的基因，包含该基因的载体，用载体转化的转化体，以及通过利用它们制备光学活性醇的方法。 本发明的特征指导从恶臭假丝酵母分离的新型多肽，编码多肽的DNA和产生多肽的转化体。 本发明的另一个特征指出通过用多肽或转化体还原羰基化合物来制备光学活性醇的方法。