ESCHERICHIA COLI EXPRESSION VECTOR AND THE USES THEREOF
    2.
    发明公开
    ESCHERICHIA COLI EXPRESSION VECTOR AND THE USES THEREOF 审中-公开
    ESCHERICHIA COLI-EXPRESSIONSVEKTOR UND VERWENDUNGEN DAVON

    公开(公告)号:EP1925670A1

    公开(公告)日:2008-05-28

    申请号:EP06775269.1

    申请日:2006-08-02

    CPC分类号: C12N15/70 C12P21/02

    摘要: The present invention discloses Escherichia coli expression vector, as well as a method for steadily, high-efficiently expressing a required target protein in E. coli using the expression vector. The expression vector is constructed from vector pRSET-A, wherein ampicillin resistance gene in said vector pRSET-A is replaced by kanamycin resistance gene. The expression vector can be used to high-efficiently express enzymes required for the production of 7-aminocephalosporanic acid, meanwhile it does not produce β-lactamase by which the productivity of 7-aminocephalosporanic acid is reduced. The present invention also discloses two expression vectors, i.e., one expression vector pHS-GHA whose gene product is D-amino acid oxidase mutant GHA, and another expression vector pT7-kan-ACY whose gene product is glutaryl-7-aminocephalosporanic acid acylase of Pseudomonas sp. SE83.

    摘要翻译: 本发明公开了大肠杆菌表达载体,以及使用表达载体在大肠杆菌中稳定地高效表达所需靶蛋白的方法。 表达载体由载体pRSET-A构建,其中所述载体pRSET-A中的氨苄青霉素抗性基因被卡那霉素抗性基因替代。 表达载体可用于高效表达生产7-氨基头孢烷酸所需的酶,同时不产生β-内酰胺酶,其中7-氨基头孢烷酸的产率降低。 本发明还公开了两种表达载体,即其基因产物是D-氨基酸氧化酶突变体GHA的一种表达载体pHS-GHA和其基因产物是戊二酰-7-氨基头孢烷酸酰基转移酶的另一表达载体pT7-kan-ACY 假单胞菌属 SE83。

    ENZYMATIC PROCESS FOR PREPARING CEPHALOSPORANIC ACID DERIVATIVES USING ALPHA-KETOACID DERIVATIVES
    3.
    发明公开
    ENZYMATIC PROCESS FOR PREPARING CEPHALOSPORANIC ACID DERIVATIVES USING ALPHA-KETOACID DERIVATIVES 审中-公开
    用α-酮酸衍生物制备头孢烯酸衍生物的酶法

    公开(公告)号:EP1379531A2

    公开(公告)日:2004-01-14

    申请号:EP02735276.4

    申请日:2002-04-18

    CPC分类号: C07D501/00

    摘要: A process for preparing cephalosporanic acid derivatives comprises the steps of enzymatically converting a 3-thiolated cephalosporin C compound of formula (III) into a 3-thiolated-α-ketoadipyl-7-aminocephalosporanic acid derivative of formula (IV), wherein R is a heterocyclic group comprising at least a nitrogen atom. Compounds of formula (IV) are used in the preparation of cephalosporin C antibiotics and derivatives thereof.

    摘要翻译: 制备头孢菌酸衍生物的方法包括以下步骤:将式(III)的3-硫醇化头孢菌素C化合物酶促转化成式(IV)的3-硫醇化-α-酮基己二酰-7-氨基头孢菌酸衍生物,其中R是 包含至少一个氮原子的杂环基团。 式(IV)化合物用于制备头孢菌素C抗生素及其衍生物。

    Purified deacetoxycephalosporin C synthase
    6.
    发明公开
    Purified deacetoxycephalosporin C synthase 失效
    Gereinigte脱乙酰氧头孢菌素-C合酶。

    公开(公告)号:EP0366354A2

    公开(公告)日:1990-05-02

    申请号:EP89310744.1

    申请日:1989-10-19

    IPC分类号: C12N9/00 C12P35/06

    摘要: Deacetoxycephalosporin C synthase obtained from cell-free extracts of Streptomyces clavuligerus was purified to near homogeneity and characterized in inactive form. The amino acid sequence was determined and the synthase recombinantly reproduced in active form in E. coli . The recombinantly produced synthase is provided in about 97% pure form by a chromatographic process. The synthase lacks DAOC hydroxylase function; however, it possesses the ability to transform 3-exo­methylenecephalosporin C to deacetylcephalosporin C.

    摘要翻译: 从无链球菌链霉菌(Streptomyces clavuligerus)的无细胞提取物获得的脱乙酰氧基头孢菌素C合酶被纯化至接近均匀性并以无活性形式表征。 测定氨基酸序列,并以大肠杆菌中活性形式重组重组合成酶。 重组产生的合成酶以约97%纯的形式通过色谱法提供。 合成酶缺乏DAOC羟化酶功能; 然而,它具有将3-甲磺酰头孢菌素C转化为脱乙酰头孢菌素C的能力。

    Biosynthesis of unnatural cephalosporins
    7.
    发明公开
    Biosynthesis of unnatural cephalosporins 失效
    非天然头孢菌素的生物合成

    公开(公告)号:EP0295724A3

    公开(公告)日:1990-04-18

    申请号:EP88200131.6

    申请日:1983-08-12

    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 1 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 - 半胱氨酰 - 缬氨酸)类型的肽前体(其中缬氨酸部分可以被任何容易获得的氨基酸置换)可以转化为其中R1是H,低级烷基或官能化羧基类型的头孢菌素和 R2是H或低级烷基,通过使前体与从原核生物如链霉菌(Streptomyces clavuligerus)的无细胞提取物分离的新型环化酶,差向异构酶和环扩张酶反应。 三种新型酶可以分别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。

    Verfahren zur Gewinnung von Cephalosporin C, seinen Salzen und Derivaten aus Kulturfiltraten oder -lösungen
    9.
    发明公开
    Verfahren zur Gewinnung von Cephalosporin C, seinen Salzen und Derivaten aus Kulturfiltraten oder -lösungen 失效
    一种用于从培养物或溶液获得的头孢菌素C,其盐和衍生物的方法。

    公开(公告)号:EP0001797A1

    公开(公告)日:1979-05-16

    申请号:EP78101206.7

    申请日:1978-10-24

    IPC分类号: C12P35/06 C07D501/12

    CPC分类号: C07D501/12

    摘要: Verfahren zur Gewinnung von Cephalosporin C, seiner Salze und Derivate aus Kulturfiltraten oder -lösungen, das darin besteht, daß man das Kulturfiltrat oder die Kulturlösung trocknet, in einem gegebenenfalls wasserhaltigen, von Wasser verschiedenen Lösungsmittel aufnimmt, und a) ein Derivat oer Aminogruppe des Cephalosporin C bildet und dieses als freie Säure oder Salz isoliert oder b) das Cephalosporin C direkt oder als schwerlösliches Salz ausfällt.

    摘要翻译: 解决方案,一种用于从培养物或其中包括在干燥培养滤液或培养液中获得的头孢菌素C,其盐和衍生物的方法,接收在任选除水溶剂以外,和a)一种衍生物OER氨基的头孢菌素的含水 ç形式和此分离为游离酸或盐,或b)的头孢菌素C直接或作为微溶的盐沉淀。