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    ENZYMATIC PROCESS FOR THE PREPARATION OF CEPHALOSPORANIC 7$b(g)-(4-CARBOXYBUTANAMIDE) ACID BY MEANS OF THE MODIFIED ENZYME D-AMINOACID OXIDASE OF $i(TRIGONOPSIS VARIABILIS) PRODUCED IN $i(ESCHERICHIA COLI)
    52.
    发明公开
    ENZYMATIC PROCESS FOR THE PREPARATION OF CEPHALOSPORANIC 7$b(g)-(4-CARBOXYBUTANAMIDE) ACID BY MEANS OF THE MODIFIED ENZYME D-AMINOACID OXIDASE OF $i(TRIGONOPSIS VARIABILIS) PRODUCED IN $i(ESCHERICHIA COLI) 审中-公开
    酶促过程用于产生修正后的dAMINOSÄUREOXIDASEENZYMS头孢-7-β-4-羧丁基酰胺MEANS在大肠杆菌中生产什么

    公开(公告)号：EP0969088A1

    公开(公告)日：2000-01-05

    申请号：EP98943898.1

    申请日：1998-09-24

    申请人： Antibioticos, S.A.

    发明人： GARCIA LOPEZ, José Luis ,  CORTES RUBIO, Estrella ,  ALONSO PALACIOS, Jorge ,  MELLADO DURAN, Encarnación ,  DIEZ GARCIA, Bruno ,  GUISAN SEIJAS, José Manuel ,  SALTO MALDONADO, Francisco ,  BARREDO FUENTE, José Luis

    IPC分类号： C12N9/06

    CPC分类号： C12N9/0024 ,  C12P35/06

    摘要： Enzymatic process for the preparation of cephalosporanic 7β-(4-carboxybutanamide) acid by using the modified enzyme D-aminoacid oxidase of Trigonopsis variabilis produced in Escherichia coli . The process for the expression of the enzyme comprises: (I) isolating the DNA corresponding to the gene which codes for the enzyme D-aminoacide oxidase; (II) removing the intron which is contained in said gene; (III) inserting the DNA fragment obtained into the plasmide which is capable of replication in Escherichia coli ; (IV) fusing at the extremity 5' of the structural region of the gene a synthetic assembler which contains a nucleotide sequence which codes for six histidines; (V) transforming a strain of Escherichia coli with the resulting recombinant plasmide; (VI) cultivating the transformed cells of Escherichia coli ; and (VII) recovering the enzyme D-aminoacid oxidase of the former cultivation operation through affinity chromatography.

    摘要翻译： （4-羧丁基酰胺），通过使用经修改的酶T的D-氨基酸氧化酶变异在大肠杆菌中产生的酸 - 对头孢菌素7测试的制备酶促过程。 对于酶的表达的方法，包括：（i）分离所述DNA对应于基因，其对于酶D-aminoacide氧化酶码; （II）除去内含子的所有其被包含在所述基因; （III）将所述DNA片段插入得到的所有质粒，它能够在大肠杆菌中复制的; （IV）在末端5“基因合成汇编其中包含的核苷酸序列的结构区域的熔化这对于六个组氨酸码; （V）转化大肠杆菌用所得的重组质粒的菌株; （VI）培养大肠杆菌的转化的细胞; 和（VII）回收前通过培养亲和层析操作的酶的D-氨基酸氧化酶。

    Biosynthesis of unnatural cephalosporins
    58.
    发明公开
    Biosynthesis of unnatural cephalosporins 失效
    生物合成on b b en en en。。。。。。

    公开(公告)号：EP0296641A2

    公开(公告)日：1988-12-28

    申请号：EP88200129.0

    申请日：1983-08-12

    申请人： QUEEN'S UNIVERSITY AT KINGSTON

    发明人： Wolfe, Saul R. R. ,  Westlake, Donald ,  Jenson, Susan

    IPC分类号： C12P35/00 ,  C12P35/06 ,  C12P37/00 ,  C12N11/18 ,  C12N9/00 ,  C12N9/90

    CPC分类号： C07K5/06139 ,  C12N9/00 ,  C12N9/90 ,  C12N11/18 ,  C12P35/00 ,  C12P35/06 ,  C12P37/00 ,  Y10S435/814 ,  Y10S435/815 ,  Y10S435/816 ,  Y10S435/886

    摘要： A peptide precursor of the ACV (aminoadipyl-cysteinyl-valine) type, in which the valine moiety may be replaced by any readily available amino acid may be converted into a cephalosporin of the type
    where R, is H, lower alkyl or functionalized carboxylic group and R 2 is H or lower alkyl, by reacting the precursor with novel cyclase, epimerase and ring expansion enzymes isolated from a cell-free extract of a prokaryotic organism such as Streptomyces clavuligerus. The three novel enzymes may be immobilized separately or together on a suitable support medium and the conversion may be carried out in a continuous mode.

    摘要翻译： ACV（氨基己二酰基 - 半胱氨酰 - 缬氨酸）类型的肽前体，其中缬氨酸部分可被任何容易获得的氨基酸替代，可转化为类型的头孢菌素，其中R 1为H，低级烷基或官能化 羧酸基团，并且R 2是H或低级烷基，通过使前体与从原核生物例如Streptomyces clavuligerus的无细胞提取物分离的新型环化酶，差向异构酶和环扩增酶反应。 三种新型酶可以单独地或一起固定在合适的载体介质上，并且转化可以以连续模式进行。

    Biosynthesis of unnatural cephalosporins
    60.
    发明公开
    Biosynthesis of unnatural cephalosporins 失效
    生物化学不对称头孢菌素。

    公开(公告)号：EP0144170A1

    公开(公告)日：1985-06-12

    申请号：EP84307731.4

    申请日：1984-11-08

    申请人： QUEEN'S UNIVERSITY AT KINGSTON

    发明人： Wolfe, Saul ,  Westlake, Donald ,  Jensen, Susan

    IPC分类号： C12P37/00 ,  C12P35/00 ,  C12P35/06 ,  C07K5/06 ,  C07D501/22 ,  A61K31/545

    CPC分类号： C07K5/06139 ,  A61K38/00 ,  C12N9/00 ,  C12P35/00 ,  C12P35/06 ,  C12P37/00

    摘要： A process for producing unnatural penicillins and cephalosporin derivatives thereof in which peptide analogs of ACV in which the L-a-aminodipyl moiety is replaced by L-S-carboxymethyl cysteine or other substituents, are reacted with cyclase, epimerase and ring expansion enzyme isolated from a cell free extract of a prokaryotic organism such as S. clavuligerus. The product depends upon the presence or absence of co-factors such as ferrous ion, a-ketoglutarate, and ascorbate. In an alternative embodiment, a penicillin analog having the formula
    may be reacted with L-cysteine to produce an analog of isopenicillin N which may be reacted with the enzyme reagent to produce the desired penicillin or cephalosporin.

    摘要翻译： 一种生产非天然青霉素和头孢菌素衍生物的方法，其中将由L-羧甲基半胱氨酸或其它取代基取代的L-氨基脯氨酰部分的ACV的肽类似物与从细胞分离的环化酶，差向异构酶和环膨化酶反应 游离提取物的原核生物，如克拉维氏梭菌。 该产品取决于辅助因子如亚铁离子，α-酮戊二酸和抗坏血酸的存在或不存在。 在替代实施方案中，具有式CHEM的青霉素类似物可以与L-半胱氨酸反应以产生异青霉素N的类似物，其可与酶试剂反应以产生所需的青霉素或头孢菌素。