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    Deacetoxycephalosporin C hydroxylase
    5.
    发明公开
    Deacetoxycephalosporin C hydroxylase 失效
    脱乙酰-C-水解酶。

    公开(公告)号：EP0420562A1

    公开(公告)日：1991-04-03

    申请号：EP90310447.9

    申请日：1990-09-25

    申请人： ELI LILLY AND COMPANY

    发明人： Yeh, Wu-Kuang ,  Dotzlaf, Joe Edward

    IPC分类号： C12N9/02 ,  C07K14/36

    CPC分类号： C12N9/00 ,  C12P35/06

    摘要： Deacetoxycephalosporin C hydroxylase is obtained in purified form from crude cell-free extracts via chromatography over a weak anionic exchange resin, ammonium sulfate fractionation, gel filtration, hydroxyl­apatite chromatography, and FPLC. The enzyme is obtained in >90% purity by further gel filtration and a second FPLC. The 28-residue amino-terminal sequence of hydroxylase is provided as well as a 9-residue amino-­terminal sequence of an internal sequence and a 3-residue carboxy-terminal sequence. In addition to the efficient conversion of DAOC to DAC, the hydroxylase provided herein converts 7β-(α-amino­adipamido)-3-exomethylenecepham-4-carboxylic acid to DAC.

    摘要翻译： 脱乙酰氧基头孢菌素C羟化酶通过弱阴离子交换树脂，硫酸铵分级分离，凝胶过滤，羟基磷灰石层析和FPLC通过色谱法从粗制无细胞提取物中纯化得到。 通过进一步的凝胶过滤和第二次FPLC获得> 90％纯度的酶。 提供羟化酶的28残基氨基末端序列以及内部序列和3-残基羧基末端序列的9残基氨基末端序列。 除了将DAOC有效转化为DAC之外，本文提供的羟化酶将7β-（α-氨基己二酰氨基）-3-外亚甲基头孢-4-羧酸转化为DAC。

    Biosynthesis of unnatural cephalosporins
    6.
    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

    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 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。

    MUTATED CEPHALOSPORIN HYDROXYLASE AND ITS APPLICATION IN DEACETYLCEPHALOSPORANIC ACID SYNTHESIS
    8.
    发明公开
    MUTATED CEPHALOSPORIN HYDROXYLASE AND ITS APPLICATION IN DEACETYLCEPHALOSPORANIC ACID SYNTHESIS 有权
    突变头孢菌素羟化酶及其在脱氧肉桂酸合成中的应用

    公开(公告)号：EP2802661A1

    公开(公告)日：2014-11-19

    申请号：EP13735852.9

    申请日：2013-01-09

    申请人： Orchid Chemicals and Pharmaceuticals Ltd

    发明人： DURAIRAAJ,Micheal ,  THIRUMOORTHY, Ramanan ,  MISHRA, Kanhu Charan ,  CHINNATHAMBI, Thangadurai ,  KRISHNAN, Cavery Manian ,  RAJASEKARAN, Padma ,  SUBRAMANI, Sugumar ,  SELVARAJ, Kavitha Daffrose ,  BALAKRISHNAN, Nataraj ,  RAVIKUMAR CHAKRAVARTHY, Sathish ,  NATRAJAN MADHIYAZHAGAN, Arulmozhi

    IPC分类号： C12N15/53 ,  C12N9/02 ,  C12P35/00

    CPC分类号： C12P35/00 ,  C12N9/0071 ,  C12P35/06 ,  Y02P20/52

    摘要： The present invention relates to a mutant hydroxylase with increased activity and greater substrate specificity towards phenylacetyl-7-ADCA derivatives for the production of phenylacetyl-7-HACA derivatives, which carries one or more amino acid modification at residue positions when compared with the wild type hydroxylase from the following group of residues, Proline at position 7, Alanine at position 40, Threonine 51, Methionine at position 53, Glutamic acid at position 82, Arginine at position 91, Threonine at position 96, Glycine at position 108, Isoleucine at position 149, Valine at position 171, Alanine at position 177, Arginine at position 182, Methioinine at position 184, Isoleucine at position 193, Phenylalanine at position 195, Glutamine at position 209, Alanine at position 210, Valine at position 226, Methionine at position 233, Leucine at position 236, Alanine at position 237, Alanine at position 241, Valine at position 249, Arginine at position 250, Serine at position 251, Glycine at position 255, Glutamic Acid at position 258, Serine at position 260, Phenylalanine at position 267, Alanine at position 280, Valine at position 307 and Asparagine at position 313.. The invention further provides a process for the preparation of deacetyl cephalosporanic acid from the corresponding deacetoxy cephalosporanic acid using an enzyme of the present invention. The invention also provides the method for producing and processing of such enzymes.

    摘要翻译： 本发明涉及用于生产苯乙酰-7-HACA衍生物的苯乙酰-7-ADCA衍生物具有增加的活性和更高的底物特异性的突变羟化酶，所述苯乙酰-7-HACA衍生物与野生型相比在残基位置携带一个或多个氨基酸修饰 来自以下残基组的羟化酶：7位脯氨酸，40位丙氨酸，苏氨酸51位，53位甲硫氨酸，82位谷氨酸，91位精氨酸，96位苏氨酸，108位甘氨酸，位置异亮氨酸 149位，171位缬氨酸，177位丙氨酸，182位精氨酸，184位甲硫氨酸，193位异亮氨酸，195位苯丙氨酸，209位谷氨酰胺，210位丙氨酸，210位缬氨酸，226位缬氨酸 233位，236位亮氨酸，237位丙氨酸，241位丙氨酸，249位缬氨酸，250位精氨酸，251位丝氨酸，G 本发明进一步提供了一种制备脱乙酰头孢菌酸的方法，该方法包括：在第255位的甘氨酸，第255位的甘氨酸，第258位的谷氨酸，第260位的丝氨酸，第267位的苯丙氨酸，第280位的丙氨酸，第307位的缬氨酸和第313位的天冬酰胺。 从相应的脱乙酸头孢菌酸使用本发明的酶。 本发明还提供了用于生产和加工这种酶的方法。