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    Cephalosporin C acylase
    1.
    发明授权
    Cephalosporin C acylase 失效
    头孢菌素C酰基转移酶

    公开(公告)号：US5336613A

    公开(公告)日：1994-08-09

    申请号：US19870

    申请日：1993-02-19

    申请人： Mineo Niwa ,  Yoshimasa Saito ,  Hitoshi Sasaki ,  Yoshinori Ishii

    发明人： Mineo Niwa ,  Yoshimasa Saito ,  Hitoshi Sasaki ,  Yoshinori Ishii

    IPC分类号： C12N1/21 ,  C12N9/80 ,  C12N15/09 ,  C12N15/55 ,  C12P35/02 ,  C12P35/06 ,  C12R1/19 ,  C12N15/52

    CPC分类号： C12P35/02 ,  C12N9/80

    摘要： The present invention concerns a mutant cephalosporin C acylase derived from a precursor of the formula:A.sup.1-268 --X.sup.1 --Tyr--X.sup.2 --A.sup.272-304 --X.sup.3 --A.sup.306-773(SEQ ID NO:1), wherein:A.sup.1-268 is the same amino acid sequence as that from Thr.sup.1 to Gly.sup.268 of native CC acylase,A.sup.272-304 is the same amino acid sequence as that from Gln.sup.272 to Tyr.sup.304 of native CC acylase,A.sup.306-773 is the same amino acid sequence as that from Val.sup.306 to Ala.sup.773 of native CC acylase,X.sup.1 is Met or other amino acid,X.sup.2 is Ala or Tyr, andX.sup.3 is Cys or Ser,provided that when X.sup.1 is Met and X.sup.2 is Ala, X.sup.3 is Ser; and that the mutant cephalosporin C acylase has a property selected from the group consisting of higher enzymatic potency and higher processing efficiency, as compared to native CC acylase. The present invention also concerns DNA encoding the mutant cephalosporin C acylase, an expression vector containing the DNA, a host cell transformed with the expression vector, a process for producing the mutant cephalosporin C acylase by culturing the transformed host cell, and a process for preparing a cephalosporin C using the mutant cephalosporin C acylase.

    摘要翻译： 本发明涉及衍生自下式的前体的突变型头孢菌素C酰基转移酶：A1-268-X1-Tyr-X2-A272-304-X3-A306-773（SEQ ID NO：1），其中：A1-268是 与天然CC酰基转移酶A272-304的Thr1至Gly268相同的氨基酸序列与天然CC酰基转移酶从Gln272至Tyr304的氨基酸序列相同，氨基酸序列与Val306至 天然CC酰基转移酶的Ala773，X1是Met或其他氨基酸，X2是Ala或Tyr，X3是Cys或Ser，条件是当X1是Met，X2是Ala时，X3是Ser; 并且与天然CC酰基转移酶相比，突变型头孢菌素C酰基转移酶具有选自较高酶效力和较高加工效率的性质。 本发明还涉及编码突变型头孢菌素C酰基转移酶的DNA，含有DNA的表达载体，用表达载体转化的宿主细胞，通过培养转化的宿主细胞产生突变型头孢菌素C酰基转移酶的方法，以及制备 使用突变型头孢菌素C酰基转移酶的头孢菌素C。

    Method for producing 2-keto-L-gulonic acid
    3.
    发明授权
    Method for producing 2-keto-L-gulonic acid 失效
    2-酮-L-古洛糖酸的制备方法

    公开(公告)号：US5834263A

    公开(公告)日：1998-11-10

    申请号：US696834

    申请日：1996-09-24

    申请人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Hayashi

    发明人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Hayashi

    IPC分类号： C12N9/02 ,  C12N9/04 ,  C12N15/53 ,  C12P7/60 ,  C12N1/00 ,  C12N15/00

    CPC分类号： C12N9/0008 ,  C12N9/0006 ,  C12P7/60 ,  Y10S435/822

    摘要： An expression vector containing both a DNA encoding an L-sorbose dehydrogenase and a DNA encoding an L-sorbosone dehydrogenase; a transformant having an ability to produce 2-keto-L-gulonic acid (hereinafter 2KLGA) at high yields from D-sorbitol, which is prepared by transforming, with said expression vector, a microorganism capable of producing L-sorbose at high yields from D-sorbitol, which has no or low 2KLGA-decomposing activity or a host microorganism having, in addition to the above-mentioned properties, no or low L-idonic acid-producing activity; and a process for producing 2KLGA, which comprises culturing said transformant in a medium containing D-sorbitol. According to the present invention, 2KLGA useful for the production of L-ascorbic acid can be produced with ease and in larger amounts by a single operation of culture.

    摘要翻译： PCT No.PCT / JP95 / 00285 Sec。 371日期：1996年9月24日 102（e）1996年9月24日PCT提交1995年2月24日PCT公布。 WO95 / 23220 PCT出版物 日期1995年8月31日含有编码L-山梨糖脱氢酶的DNA和编码L-山梨糖酮脱氢酶的DNA的表达载体; 具有从D-山梨醇以高产率生产2-酮-L-古洛糖酸（以下称为2KLGA）的能力的转化体，其通过用所述表达载体转化能以高产率生产L-山梨糖的微生物制备 没有或低的2KLGA分解活性的D-山梨糖醇或除了上述性质之外，没有或低的L-天冬糖酸生产活性的宿主微生物; 以及生产2KLGA的方法，其包括在含有D-山梨糖醇的培养基中培养所述转化体。 根据本发明，用于生产L-抗坏血酸的2KLGA可以通过单次的培养操作容易地和大量地生产。

    L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from gluconobacter oxydans T-100
    4.
    发明授权
    L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from gluconobacter oxydans T-100 失效
    L-山梨糖脱氢酶和由氧化葡萄糖葡萄球菌T-100获得的新型L-山梨糖酮脱氢酶

    公开(公告)号：US5753481A

    公开(公告)日：1998-05-19

    申请号：US513841

    申请日：1995-11-01

    申请人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Suzuki

    发明人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Suzuki

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

    CPC分类号： C12N9/0006 ,  Y10S435/822

    摘要： A novel L-sorbose dehydrogenase (SDH) and a novel L-sorbosone dehydrogenase both derived from Gluconobacter oxydans T-100, a DNA which encodes the SDH and/or SNDH, an expression vector which contains the DNA, a host cell transformed by the expression vector and a process for producing the SDH and/or SNDH, which comprises culturing the host cell in a medium and recovering the SDH and/or SNDH from the resulting culture. The SDH and SNDH of the present invention are useful enzymes having preferable properties for the production of 2-keto-L-gulonic acid, as well as L-ascorbic acid. According to the production method of the present invention, the SDH and SNDH having such preferable properties can be produced in large amounts by genetic engineering.

    摘要翻译： PCT No.PCT / JP94 / 00369 Sec。 371日期：1995年11月1日 102（e）日期1995年11月1日PCT 1994年3月8日PCT公布。 公开号WO94 / 20609 日期1994年9月15日新型L-山梨糖脱氢酶（SDH）和新型L-山梨糖酮脱氢酶均来自氧化葡糖杆菌T-100，编码SDH和/或SNDH的DNA，含有DNA的表达载体， 由表达载体转化的宿主细胞和产生SDH和/或SNDH的方法，其包括在培养基中培养宿主细胞并从所得培养物中回收SDH和/或SNDH。 本发明的SDH和SNDH是生产2-酮-L-古洛糖酸以及L-抗坏血酸的优选性质的有用的酶。 根据本发明的制造方法，可以通过遗传工程大量生产具有优选性质的SDH和SNDH。

    L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from gluconobacter oxydans T-100
    5.
    发明授权
    L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from gluconobacter oxydans T-100 失效
    L-山梨糖脱氢酶和由氧化葡萄糖葡萄球菌T-100获得的新型L-山梨糖酮脱氢酶

    公开(公告)号：US06197562B1

    公开(公告)日：2001-03-06

    申请号：US09118317

    申请日：1998-07-17

    申请人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Suzuki

    发明人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Suzuki

    IPC分类号： C12N904

    CPC分类号： C12N9/0006 ,  Y10S435/822

    摘要： A novel L-sorbose dehydrogenase (SDH) and a novel L-sorbosone dehydrogenase both derived from Gluconobacter oxydans T-100, a DNA which encodes the SDH and/or SNDH, an expression vector which contains the DNA, a host cell transformed by the expression vector and a process for producing the SDH and/or SNDH, which comprises culturing the host cell in a medium and recovering the SDH and/or SNDH from the resulting culture. The SDH and SNDH of the present invention are useful enzymes having preferable properties for the production of 2-keto-L-gulonic acid, as well as L-ascorbic acid. According to the production method of the present invention, the SDH and SNDH having such preferable properties can be produced in large amounts by genetic engineering.

    摘要翻译： 一种新型的L-山梨糖脱氢酶（SDH）和一种新型的L-山梨糖酮脱氢酶，它们都衍生自氧化葡糖杆菌T-100，一种编码SDH和/或SNDH的DNA，含有该DNA的表达载体，由 表达载体和产生SDH和/或SNDH的方法，其包括在培养基中培养宿主细胞并从所得培养物中回收SDH和/或SNDH。 本发明的SDH和SNDH是生产2-酮-L-古洛糖酸以及L-抗坏血酸的优选性质的有用的酶。 根据本发明的制造方法，可以通过遗传工程大量生产具有优选性能的SDH和SNDH。

    Method for producing 2-keto-L-gulonic acid
    7.
    发明授权
    Method for producing 2-keto-L-gulonic acid 失效
    2-酮-L-古洛糖酸的制备方法

    公开(公告)号：US5888786A

    公开(公告)日：1999-03-30

    申请号：US943285

    申请日：1997-10-03

    申请人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Hayashi

    发明人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Hayashi

    IPC分类号： C12N9/02 ,  C12N9/04 ,  C12N15/53 ,  C12P7/60 ,  C12N1/00 ,  C12N15/00

    CPC分类号： C12N9/0008 ,  C12N9/0006 ,  C12P7/60 ,  Y10S435/822

    摘要： An expression vector containing both a DNA encoding an L-sorbose dehydrogenase and a DNA encoding an L-sorbosone dehydrogenase; a transformant having an ability to produce 2-keto-L-gulonic acid (hereinafter 2KLGA) at high yields from D-sorbitol, which is prepared by transforming, with said expression vector, a microorganism capable of producing L-sorbose at high yields from D-sorbitol, which has no or low 2KLGA-decomposing activity or a host microorganism having, in addition to the above-mentioned properties, no or low L-idonic acid-producing activity; and a process for producing 2KLGA, which comprises culturing said transformant in a medium containing D-sorbitol. According to the present invention, 2KLGA useful for the production of L-ascorbic acid can be produced with ease and in larger amounts by a single operation of culture.

    摘要翻译： 含有编码L-山梨糖脱氢酶的DNA和编码L-山梨糖酮脱氢酶的DNA的表达载体; 具有从D-山梨醇以高产率生产2-酮-L-古洛糖酸（以下称为2KLGA）的能力的转化体，其通过用所述表达载体转化能以高产率生产L-山梨糖的微生物制备 没有或低的2KLGA分解活性的D-山梨糖醇或除了上述性质之外，没有或低的L-天冬糖酸生产活性的宿主微生物; 以及生产2KLGA的方法，其包括在含有D-山梨糖醇的培养基中培养所述转化体。 根据本发明，用于生产L-抗坏血酸的2KLGA可以通过单次的培养操作容易地和大量地生产。

    L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from Gluconobacter oxydans T-100
    8.
    发明授权
    L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from Gluconobacter oxydans T-100 失效
    L-山梨糖脱氢酶和由氧化葡糖杆菌T-100获得的新型L-山梨糖酮脱氢酶

    公开(公告)号：US5861292A

    公开(公告)日：1999-01-19

    申请号：US942673

    申请日：1997-10-02

    申请人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Suzuki

    发明人： Mineo Niwa ,  Yoshimasa Saito ,  Yoshinori Ishii ,  Masaru Yoshida ,  Hiromi Suzuki

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

    CPC分类号： C12N9/0006 ,  Y10S435/822

    摘要： A novel L-sorbose dehydrogenase (SDH) and a novel L-sorbosone dehydrogenase (SNDH) both derived from Gluconobacter oxydans T-100, a DNA which encodes the SDH and/or SNDH, an expression vector which contains the DNA, a host cell transformed by the expression vector and a process for producing the SDH and/or SNDH, which comprises culturing the host cell in a medium and recovering the SDH and/or SNDH from the resulting culture. The SDH and SNDH of the present invention are useful enzymes having preferable properties for the production of 2-keto-L-gulonic acid, as well as L-ascorbic acid. According to the production method of the present invention, the SDH and SNDH having such preferable properties can be produced in large amounts by genetic engineering.

    摘要翻译： 来自氧化葡糖杆菌T-100的新型L-山梨糖脱氢酶（SDH）和新型L-山梨糖酮脱氢酶（SNDH），编码SDH和/或SNDH的DNA，含有DNA的表达载体，宿主细胞 通过表达载体和产生SDH和/或SNDH的方法转化，其包括在培养基中培养宿主细胞并从所得培养物中回收SDH和/或SNDH。 本发明的SDH和SNDH是生产2-酮-L-古洛糖酸以及L-抗坏血酸的优选性质的有用的酶。 根据本发明的制造方法，可以通过遗传工程大量生产具有优选性能的SDH和SNDH。

    Cephalosporin C acylase
    9.
    发明授权
    Cephalosporin C acylase 失效
    头孢菌素C酰基转移酶

    公开(公告)号：US5804429A

    公开(公告)日：1998-09-08

    申请号：US633760

    申请日：1996-05-01

    申请人： Mineo Niwa ,  Yoshimasa Saito ,  Takao Fujimura ,  Yoshinori Ishii ,  Yuji Noguchi

    发明人： Mineo Niwa ,  Yoshimasa Saito ,  Takao Fujimura ,  Yoshinori Ishii ,  Yuji Noguchi

    IPC分类号： C12N15/09 ,  C12N1/21 ,  C12N9/14 ,  C12N9/80 ,  C12N15/55 ,  C12P35/02 ,  C12R1/19 ,  C07H21/04 ,  C12N15/00 ,  C12P21/06

    CPC分类号： C12P35/02 ,  C12N9/80

    摘要： A mutant CC acylase wherein at least one amino acid at the Ala.sup.49, Met.sup.164, Ser.sup.166, Met.sup.174, Glu.sup.358, Met.sup.465, Met.sup.506, or Met.sup.750 position of the amino acid sequence of the native CC acylase is replaced by a different amino acid, a DNA coding therefor, an expression vector containing the said DNA, a microorganism transformed with the said expression vector, the production of the CC acylase by culturing the said transformant, and use thereof for the production of a compound. The mutant CC acylase of the invention has desirable properties in terms of enzymatic potency, alteration of pH profile, efficiency of processing, and the like.

    摘要翻译： PCT No.PCT / JP94 / 01799 Sec。 371日期：1996年5月1日 102（e）日期1996年5月1日PCT 1994年10月26日PCT公布。 公开号WO95 / 12680 日期1995年5月11日，其中天然CC酰基转移酶的氨基酸序列的Ala49，Met164，Ser166，Met174，Glu358，Met465，Met506或Met750位置的至少一个氨基酸被不同的氨基酸替代的突变CC酰基转移酶 编码该DNA的DNA，含有上述DNA的表达载体，用上述表达载体转化的微生物，通过培养上述转化体产生CC酰基转移酶及其制备化合物的用途。 本发明的突变CC酰基转移酶在酶效力，pH曲线的改变，加工效率等方面具有期望的性质。