公开(公告)号：EP0974669A4

公开(公告)日：2000-01-26

申请号：EP97903619

申请日：1997-02-27

申请人： NIPPON SODA CO

发明人： KOBAYASHI YOICHI ,  WATABE KEN ,  OHIRA MAHITO ,  HAYAKAWA KOICHI

IPC分类号： C12N1/20 ,  C12P7/42 ,  C12R1/01

CPC分类号： C12P7/42

摘要： A process for preparing alpha -hydroxy acids represented by the general formula (II): RCH(OH)COOH (wherein R represents a hydrogen atom, an optionally substituted C1-C6 alkyl group, an optionally substituted C2-C6 alkenyl group, an optionally substituted C1-C6 alkoxy group, an optionally substituted aryl group, an optionally substituted aryloxy group, or an optionally substituted heterocyclic group) by allowing a microorganism to act on alpha -hydroxy nitriles (I): RCH(OH)CN (wherein R is as defined above) to hydrolyze and convert the alpha -hydroxy nitriles to alpha -hydroxy acids (II), wherein the alpha -hydroxy acids (II) are produced and accumulated in an aqueous solvent by a microorganism having the concentration resistance to the alpha -hydroxy nitriles (I) and/or alpha -hydroxy acids (II) and durability preferably in the presence of a cyanide, and harvested. According to this process, the use of the microorganism having the concentration resistance to the alpha -hydroxy nitriles (I) and/or alpha -hydroxy acids (II) and durability high enough to permit the activity to persist for a long period of time enables alpha -hydroxy acids (II) to be accumulated in high concentrations and cell bodies to be repeatedly used, and hence enables alpha -hydroxy acids (II) to be efficiently prepared. The addition of a cyanide to the reaction system results in more efficient preparation of alpha -hydroxy acids (II).

摘要翻译： 一种制备由通式（II）表示的α-羟基酸的方法：RCH（OH）COOH（其中R表示氢原子，任选取代的C 1 -C 6烷基，任选取代的C 2 -C 6烯基，任选取代的 取代的C 1 -C 6烷氧基，任选取代的芳基，任选取代的芳氧基或任选取代的杂环基），通过使微生物作用于α-羟基腈（I）：RCH（OH）CN 如上所述）将α-羟基腈水解并转化为α-羟酸（II），其中α-羟酸（II）通过对α - 羟基羧酸具有浓度耐受性的微生物在含水溶剂中产生并积聚， 羟基腈（I）和/或α-羟基酸（II），并优选在氰化物存在下耐久，并收获。 根据该方法，使用对α-羟基腈（I）和/或α-羟基酸（II）具有浓度耐受性的微生物并且足够高以使活性持续很长时间的耐久性能够实现 α-羟基酸（II）以高浓度聚集，细胞体重复使用，因此能够有效地制备α-羟基酸（II）。 向反应体系中加入氰化物导致更有效地制备α-羟基酸（II）。

公开(公告)号：EP0210279A4

公开(公告)日：1987-06-19

申请号：EP86900852

申请日：1986-01-24

申请人： SAGAMI CHEM RES ,  CENTRAL GLASS CO LTD ,  HODOGAYA CHEMICAL CO LTD ,  NIPPON SODA CO ,  NISSAN CHEMICAL IND LTD ,  TOYO SODA MFG CO LTD

发明人： MIYAKE TOSHIO ,  HIBINO YASUO ,  KOBAYASHI YOICHI ,  WATABE KEN ,  OMORI MUNEKI ,  MIKI TETSUZO ,  YOKOYAMA MIDORI ,  MATSUMOTO REIKO ,  WATANABE KAZUO ,  NUMAO NAGANORI

IPC分类号： C12N1/21 ,  C12N9/72 ,  C12N15/58 ,  C12N15/73 ,  C12P21/02

CPC分类号： C12N9/6462 ,  C12N15/73 ,  C12Y304/21073

摘要： Stabilized human prourokinase-like polypeptides wherein the 135th lysine from the N-terminal of human prourokinase is substituted by an amino acid other than basic amino acids, the 157th phenylalanine is substituted by an acidic amino acid, or both of the amino acids are substituted as described above; DNA segment which codes said polypeptides; plasmid which contains said DNA segment; E. coli transformed by said plasmid; and a process for preparing said polypeptide by using said E. coli. The polypeptides are stable against proteases such as plasmin, thrombin, trypsin, etc., thus being easily purified and collected in a pure form. When applied to a living body, the polypeptides are expected to provide a persistent fibrinolytic activity.