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    Recombinant decarbamylases for producing D-.alpha.-amino acids
    1.
    发明授权
    Recombinant decarbamylases for producing D-.alpha.-amino acids 失效
    用于产生D-α-氨基酸的重组脱氨基甲酰酶

    公开(公告)号：US5824522A

    公开(公告)日：1998-10-20

    申请号：US294871

    申请日：1994-08-22

    申请人： Yasuhiro Ikenaka ,  Hirokazu Nanba ,  Masayuki Takano ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Satomi Takahashi ,  Kazuma Okubo ,  Kazuhiko Yamada ,  Yoshiro Hiraishi

    发明人： Yasuhiro Ikenaka ,  Hirokazu Nanba ,  Masayuki Takano ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Satomi Takahashi ,  Kazuma Okubo ,  Kazuhiko Yamada ,  Yoshiro Hiraishi

    IPC分类号： C12N9/80 ,  C12N9/96 ,  C12N11/02 ,  C12N11/08 ,  C12P13/04 ,  C12P13/06 ,  C12P13/08 ,  C12P13/22 ,  C12P41/00 ,  C12N1/20 ,  C12N9/14 ,  C12N11/00

    CPC分类号： C12N11/02 ,  C12N11/08 ,  C12N9/80 ,  C12N9/96 ,  C12P13/04 ,  C12P13/06 ,  C12P13/08 ,  C12P13/22 ,  C12P41/009 ,  Y10S435/849 ,  Y10S435/874

    摘要： Decarbamylases are provided capable of producing D-.alpha.-amino acids by hydrolysis of N-carbamyl-D-.alpha.-amino acids. A source of the decarbamylases is recombinant microorganisms produced by gene manipulation methods. Decarbamylases having improved thermostability can be obtained in which amino acids at a thermostability-related site of a natural decarbamylase have been replaced with other amino acids by mutating a DNA fragment encoding the natural decarbamylase. Recombinant DNA is obtained from a vector DNA and a DNA fragment encoding a natural decarbamylase where the nucleic acid sequence encoding an amino acid at a thermostability-related site is replaced with a nucleic acid sequence encoding another amino acid. The recombinant DNA is used to produce transformants that produce thermostable decarbamylases. The decarbamylases may be immobilized in purified, partially-purified or crude form or in the form of decarbamylase-containing microbial cells on a support such as a polymer having ion exchange groups or covalent bonding groups. The presence of an antioxidant such as dithiothreitol results in an immobilized decarbamylase preparation that can be repeatedly used 30 times or more.

    摘要翻译： 提供的脱甲酰氨基酶能够通过N-氨基甲酰基-D-α-氨基酸的水解产生D-氨基酸。 脱氨基甲酰酶的来源是通过基因操作方法产生的重组微生物。 通过突变编码天然脱氨基甲酰酶的DNA片段，可以得到具有改善的热稳定性的脱甲酰氨基磷酸酶，其中天然脱氨基甲酰酶的热稳定性相关部位的氨基酸已被其它氨基酸替代。 从载体DNA和编码天然脱氨基甲酰酶的DNA片段获得重组DNA，其中将编码热稳定性相关部位的氨基酸的核酸序列替换为编码另一氨基酸的核酸序列。 重组DNA用于产生产生热稳定性脱氨基甲酰酶的转化体。 脱氨基甲酰酶可以固定在纯化的，部分纯化的或粗制的形式中，或以含脱氨基甲酰酶的微生物细胞的形式固定在载体上，例如具有离子交换基团或共价键合基团的聚合物。 抗氧化剂如二硫苏糖醇的存在导致可重复使用30次以上的固定化脱氨基甲酰酶制剂。

    Process for production of D-.alpha.-amino acids
    2.
    发明授权
    Process for production of D-.alpha.-amino acids 失效
    生产D-α-氨基酸的方法

    公开(公告)号：US5695968A

    公开(公告)日：1997-12-09

    申请号：US479638

    申请日：1995-06-07

    申请人： Hirokazu Nanba ,  Yukio Yamada ,  Masayuki Takano ,  Yasuhiro Ikenaka ,  Satomi Takahashi ,  Kazuyoshi Yajima

    发明人： Hirokazu Nanba ,  Yukio Yamada ,  Masayuki Takano ,  Yasuhiro Ikenaka ,  Satomi Takahashi ,  Kazuyoshi Yajima

    IPC分类号： C12N1/21 ,  C12N9/80 ,  C12N15/55 ,  C12P13/04 ,  C12P41/00 ,  C07H19/00 ,  C12N1/20 ,  C12P21/06

    CPC分类号： C12P41/009 ,  C12N9/80 ,  C12P13/04

    摘要： The present invention is directed to a gene which is related to a D-N-carbamoyl-.alpha.-amino acid amidohydrolase which is an enzyme capable of converting D-N-carbamoyl-.alpha.-amino acids into D-.alpha.-amino acids; a recombinant plasmid in which a DNA fragment containing the gene is incorporated into a vector; a microorganism belonging to the genus Escherichia, Pseudomonas, Flavobacterium, Bacillus, Serratia, Corynebacterium, or Brevibacterium, which is transformed by incorporating the recombinant plasmid thereinto; a process for the production of D-N-carbamoyl-.alpha.-amino acid amidohydrolases, comprising the steps of cultivating the transformed microorganism and collecting the desired product therefrom; a D-N-carbamoyl-.alpha.-amino acid amidohydrolase obtained by the method; and a process for the production of D-.alpha.-amino acids with the aid of an action of the enzyme. The D-N-carbamoyl-.alpha.-amino acid amidohydrolase can be fixed on a support for immobilization and used as an immobilized enzyme.

    摘要翻译： 本发明涉及与D-N-氨基甲酰基-α-氨基酸酰胺水解酶相关的基因，其是能够将D-N-氨基甲酰基-α-氨基酸转化为D-α-氨基酸的酶; 将含有该基因的DNA片段并入载体的重组质粒; 通过将重组质粒并入而转化的属于埃希氏杆菌属，假单胞菌属，黄杆菌属，芽孢杆菌属，沙雷氏菌属，棒状杆菌属或短杆菌属的微生物; 制备D-N-氨基甲酰基-α-氨基酸酰胺水解酶的方法，包括培养转化的微生物并从其中收集所需产物的步骤; 通过该方法获得的D-N-氨基甲酰基-α-氨基酸酰胺水解酶; 以及借助于酶的作用生产D-氨基酸的方法。 D-N-氨基甲酰基-α-氨基酸酰胺水解酶可以固定在用于固定的载体上并用作固定化酶。

    Process for the production of D-.alpha.-amino acids by hydrolysis of the corresponding N-carbamyl derivative
    3.
    发明授权
    Process for the production of D-.alpha.-amino acids by hydrolysis of the corresponding N-carbamyl derivative 失效
    通过水解相应的N-氨基甲酰基衍生物生产D-氨基酸的方法

    公开(公告)号：US5902736A

    公开(公告)日：1999-05-11

    申请号：US244657

    申请日：1994-06-06

    申请人： Hideaki Yamada ,  Sakayu Shimizu ,  Yasuhiro Ikenaka ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Hirokazu Nanba ,  Masayuki Takano ,  Satomi Takahashi

    发明人： Hideaki Yamada ,  Sakayu Shimizu ,  Yasuhiro Ikenaka ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Hirokazu Nanba ,  Masayuki Takano ,  Satomi Takahashi

    IPC分类号： C12P13/04 ,  C12P41/00 ,  C07C1/04

    CPC分类号： C12P13/04 ,  C12P41/009 ,  Y10S435/874 ,  Y10S435/911

    摘要： In a process for the production of a D-.alpha.-amino acid, in which an N-carbamyl-D-.alpha.-amino acid corresponding to the general formula: ##STR1## wherein R represents phenyl, hydroxy-substituted phenyl, substituted or unsubstituted alkyl, or thienyl, is converted by a microbial enzyme in an aqueous medium to a D-.alpha.-amino acid corresponding to the general formula: ##STR2## wherein R is the same as defined above, decarbamylase produced by a microorganism of the genus Comamonas, Blastobacter, Alcaligenes, Sporosarcina, Rhizobium, Bradyrhizobium or Arthrobacter is used as the enzyme converting the N-carbamyl-D-.alpha.-amino acid to the D-.alpha.-amino acid.The conversion of the N-carbamyl-D-.alpha.-amino acids to the D-.alpha.-amino acids is carried out in a neutral to alkaline pH range.

    摘要翻译： PCT No.PCT / JP93 / 01408 Sec。 371日期：1994年6月6日 102（e）日期1994年6月6日PCT提交1993年10月1日PCT公布。 公开号WO94 / 08030 日期1994年4月14日在制备D-氨基酸的方法中，其中N-氨基甲酰基-D-α-氨基酸对应于通式：其中R表示苯基，羟基取代的苯基，取代的 或未取代的烷基或噻吩基通过微生物酶在水性介质中转化成对应于以下通式的D-α-氨基酸：其中R与上述定义相同，由Comamonas属的微生物产生的脱氨基甲酰酶， 使用喷杆菌属，产碱杆菌属，孢子体菌属，根瘤菌属，Bradyrhizobium或节杆菌作为将N-氨基甲酰基-D-α-氨基酸转化为D-氨基酸的酶。 N-氨基甲酰基-D-α-氨基酸向D-α-氨基酸的转化在中性至碱性pH范围内进行。

    DNA coding for decarbamylase improved in thermostability and use thereof
    4.
    发明授权
    DNA coding for decarbamylase improved in thermostability and use thereof 失效
    编码脱氨基甲酰酶的DNA改善了热稳定性和使用

    公开(公告)号：US6083752A

    公开(公告)日：2000-07-04

    申请号：US876398

    申请日：1997-06-16

    申请人： Yasuhiro Ikenaka ,  Hirokazu Nanba ,  Masayuki Takano ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Satomi Takahashi

    发明人： Yasuhiro Ikenaka ,  Hirokazu Nanba ,  Masayuki Takano ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Satomi Takahashi

    IPC分类号： C12N9/80 ,  C12N15/55 ,  C12N15/72 ,  C12P13/06 ,  C12P13/08 ,  C12P13/22 ,  C12N15/00 ,  C12N5/10 ,  C12N9/14

    CPC分类号： C12N15/72 ,  C12N9/80 ,  C12P13/06 ,  C12P13/08 ,  C12P13/22

    摘要： A DNA fragment coding for a decarbamylase protein improved in thermostability as the result of replacement of at least one base of a DNA fragment coding for a decarbamylase protein derived from a microorganism with another base and the resultant replacement of at least one of the corresponding amino acids, and its production process; a vector containing the DNA fragment; a transformant obtained by transformation with the vector; as well as a decarbamylase improved in thermostability and its production process. Also disclosed is a process for producing a D-.alpha.-amino acid, which comprises converting an N-carbamoyl-D-.alpha.-amino acid into the corresponding D-.alpha.-amino acid in an aqueous medium by the action of a decarbamylase having a thermostable temperature of 65.degree. C. or higher; and collecting the D-.alpha.-amino acid produced.

    摘要翻译： 由于将来自微生物的脱氨基甲酰酶蛋白质的至少一个碱基替换为另一碱基的氨基酸残基的至少一个碱基，所以替代了至少一个相应的氨基酸 ，及其生产工艺; 含有DNA片段的载体; 通过用载体转化获得的转化体; 以及改进了热稳定性的脱氨基甲酰酶及其制备方法。 还公开了一种制备D-α-氨基酸的方法，该方法包括通过脱氨基甲酰酶的作用将N-氨基甲酰基-D-α-氨基酸转化成水性介质中的相应的D- 耐热温度在65℃以上; 并收集产生的D-α-氨基酸。

    Process for production of D-.alpha.-amino acids
    5.
    发明授权
    Process for production of D-.alpha.-amino acids 失效
    生产D-α-氨基酸的方法

    公开(公告)号：US5565344A

    公开(公告)日：1996-10-15

    申请号：US917111

    申请日：1992-08-07

    申请人： Hirokazu Nanba ,  Yukio Yamada ,  Masayuki Takano ,  Yasuhiro Ikenaka ,  Satomi Takahashi ,  Kazuyoshi Yajima

    发明人： Hirokazu Nanba ,  Yukio Yamada ,  Masayuki Takano ,  Yasuhiro Ikenaka ,  Satomi Takahashi ,  Kazuyoshi Yajima

    IPC分类号： C12N1/21 ,  C12N9/80 ,  C12N15/55 ,  C12P13/04 ,  C12P41/00 ,  C07H19/00 ,  C12N1/12 ,  C12N9/14

    CPC分类号： C12P41/009 ,  C12N9/80 ,  C12P13/04

    摘要： The present invention is directed to a gene which is related to a D-N-carbamoyl-.alpha.-amino acid amidohydrolase which is an enzyme capable of converting D-N-carbamoyl-.alpha.-amino acids into D-.alpha.-amino acids; a recombinant plasmid in which a DNA fragment containing the gene is incorporated into a vector; a microorganism belonging to the genus Escherichia, Pseudomonas, Flavobacterium, Bacillus, Serratia, Corynebacterium, or Brevibacterium, which is transformed by incorporating the recombinant plasmid thereinto; a process for the production of D-N-carbamoyl-.alpha.-amino acid amidohydrolases, comprising the steps of cultivating the transformed microorganism and collecting the desired product therefrom; a D-N-carbamoyl-.alpha.-amino acid amidohydrolase obtained by the method; and a process for the production of D-.alpha.-amino acids with the aid of an action of the enzyme.The D-N-carbamoyl-.alpha.-amino acid amidohydrolase can be fixed on a support for immobilization and used as an immobilized enzyme.

    摘要翻译： PCT No.PCT / JP91 / 01696 Sec。 371日期：1992年8月7日 102（e）日期1992年8月7日PCT 1991年12月6日PCT PCT。 出版物WO92 / 10579 日期1992年6月25日本发明涉及与能够将D-N-氨基甲酰基-α-氨基酸转化为D-氨基酸的酶的D-N-氨基甲酰基-α-氨基酸酰胺水解酶相关的基因。 将含有该基因的DNA片段并入载体的重组质粒; 通过将重组质粒并入而转化的属于埃希氏杆菌属，假单胞菌属，黄杆菌属，芽孢杆菌属，沙雷氏菌属，棒状杆菌属或短杆菌属的微生物; 制备D-N-氨基甲酰基-α-氨基酸酰胺水解酶的方法，包括培养转化的微生物并从其中收集所需产物的步骤; 通过该方法获得的D-N-氨基甲酰基-α-氨基酸酰胺水解酶; 以及借助于酶的作用生产D-氨基酸的方法。 D-N-氨基甲酰基-α-氨基酸酰胺水解酶可以固定在用于固定的载体上并用作固定化酶。

    Decarbamylase isolated from comamonas or blastobacter
    7.
    发明授权
    Decarbamylase isolated from comamonas or blastobacter 失效
    从comamonas或blastobacter分离的脱氨基甲酰酶

    公开(公告)号：US5863785A

    公开(公告)日：1999-01-26

    申请号：US479639

    申请日：1995-06-07

    申请人： Hideaki Yamada ,  Sakayu Shimizu ,  Yasuhiro Ikenaka ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Hirokazu Nanba ,  Masayuki Takano ,  Satomi Takahashi

    发明人： Hideaki Yamada ,  Sakayu Shimizu ,  Yasuhiro Ikenaka ,  Kazuyoshi Yajima ,  Yukio Yamada ,  Hirokazu Nanba ,  Masayuki Takano ,  Satomi Takahashi

    IPC分类号： C12P13/04 ,  C12P41/00 ,  C12N9/78 ,  C12N9/80

    CPC分类号： C12P13/04 ,  C12P41/009 ,  Y10S435/874 ,  Y10S435/911

    摘要： In a process for the production of a D-.alpha.-amino acid, in which an N-carbamyl-D-.alpha.-amino acid corresponding to the general formula: ##STR1## wherein R represents phenyl, hydroxy-substituted phenyl, substituted or unsubstituted alkyl, or thienyl, is converted by a microbial enzyme in an aqueous medium to a D-.alpha.-amino acid corresponding to the general formula: ##STR2## wherein R is the same as defined above, decarbamylase produced by a microorganism of the genus Comamonas, Blastobacter, Alcaligenes, Sporosarcina, Rhizobium, Bradyrhizobium or Arthrobacter is used as the enzyme converting the N-carbamyl-D-.alpha.-amino acid to the D-.alpha.-amino acid.The conversion of the N-carbamyl-D-.alpha.-amino acids to the D-.alpha.-amino acids is carried out in a neutral to alkaline pH range.

    摘要翻译： 在制备D-α-氨基酸的方法中，其中相应于通式为：苯基，取代或未取代的烷基或噻吩基的N-氨基甲酰基-D-α-氨基酸被转化 通过微生物酶在水性介质中与对应于以下通式的D-α-氨基酸反应：其中R与上述定义相同，由Comamonas属，Blastobacter，Alcaligenes，Sporosarcina属的微生物产生的脱氨基甲酰酶， 使用根瘤菌，Bradyrhizobium或节杆菌作为将N-氨基甲酰基-D-α-氨基酸转化为D-α-氨基酸的酶。 N-氨基甲酰基-D-α-氨基酸向D-α-氨基酸的转化在中性至碱性pH范围内进行。

    Process for producing coenzyme Q10
    8.
    发明授权
    Process for producing coenzyme Q10 有权
    生产辅酶Q10的方法

    公开(公告)号：US06762037B1

    公开(公告)日：2004-07-13

    申请号：US09830111

    申请日：2001-07-23

    申请人： Hideyuki Matsuda ,  Makoto Kawamukai ,  Kazuyoshi Yajima ,  Yasuhiro Ikenaka ,  Junzo Hasegawa ,  Satomi Takahashi

    发明人： Hideyuki Matsuda ,  Makoto Kawamukai ,  Kazuyoshi Yajima ,  Yasuhiro Ikenaka ,  Junzo Hasegawa ,  Satomi Takahashi

    IPC分类号： C12P2102

    CPC分类号： C12N9/1085 ,  C12P7/66

    摘要： The present invention has for its object to isolate a gene coding for the enzyme synthesizing the coenzyme Q10 side chain from a fungal strain of the genus Saitoella and exploit it to advantage for the efficient microbial production of coenzyme Q10. The present invention provides; a DNA having the nucleotide sequence shown under SEQ ID NO:1 a DNA having a nucleotide sequence derived from the nucleotide sequence of SEQ ID NO:1 by the deletion, addition, insertion and/or substitution of one or a plurality of nucleotides and coding for a protein having decaprenyl diphosphate synthase activity a DNA which hybridizes with the DNA having the nucleotide sequence of SEQ ID NO:1 under a stringent condition and codes for a protein having decaprenyl diphosphate synthase activity.

    摘要翻译： 本发明的目的是从编码斋藤本的真菌菌株中分离编码合成辅酶Q10侧链的酶的基因，并有利于辅酶Q10的有效微生物产生。本发明提供了一种DNA 具有SEQ ID NO：1a所示的核苷酸序列，其具有通过删除，添加，插入和/或取代一个或多个核苷酸而编码具有SEQ ID NO：1的核苷酸序列的核苷酸序列的DNA，并编码具有SEQ ID NO： 癸酰二磷酸合酶活性a在严格条件下与具有SEQ ID NO：1的核苷酸序列的DNA杂交的DNA，并编码具有十聚丙二酰二磷酸合成酶活性的蛋白质的DNA。