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    Mutant penicillin G acylases
    1.
    发明授权
    Mutant penicillin G acylases 有权
    突变型青霉素G酰基转移酶

    公开(公告)号：US08541199B2

    公开(公告)日：2013-09-24

    申请号：US13141877

    申请日：2009-12-22

    申请人： Jan Metske Van Der Laan ,  Harold Monro Moody ,  Richard Kerkman ,  Thomas Van Der Does

    发明人： Jan Metske Van Der Laan ,  Harold Monro Moody ,  Richard Kerkman ,  Thomas Van Der Does

    IPC分类号： C12P1/00 ,  C12N9/84 ,  C12N1/20 ,  C07H21/04

    CPC分类号： C12N9/84 ,  C12P35/04 ,  C12P37/04

    摘要： The present invention relates to a mutant prokaryotic penicillin G acylase derived from a wild-type penicillin G acylase characterized in that the mutant is having an amino acid substitution at one or more amino acid positions selected from the group consisting of amino acid positions A3, A77, A90, A144 A192, B24, B109, B148, B313, B460 and B488 according to the amino acid numbering of the Escherichia coli penicillin G acylase having the amino acid sequence depicted in SEQ ID No: 1.

    摘要翻译： 本发明涉及源自野生型青霉素G酰基转移酶的突变型原核青霉素G酰基转移酶，其特征在于突变体在选自氨基酸位置A3，A77的一个或多个氨基酸位置具有氨基酸取代 ，A90，A144A192，B24，B109，B148，B313，B460和B488，其具有SEQ ID No：1所示的氨基酸序列的大肠杆菌青霉素G酰基转移酶的氨基酸编号。

    Novel acylase gene
    3.
    发明申请
    Novel acylase gene 失效
    新型酰基转移酶基因

    公开(公告)号：US20060035363A1

    公开(公告)日：2006-02-16

    申请号：US10516587

    申请日：2003-05-30

    申请人： Akiko Nishi ,  Takumi Mano ,  Shinichi Yokota ,  Masayuki Takano ,  Kazuyoshi Yajima

    发明人： Akiko Nishi ,  Takumi Mano ,  Shinichi Yokota ,  Masayuki Takano ,  Kazuyoshi Yajima

    IPC分类号： C12N9/80 ,  C07H21/04 ,  C12P21/06 ,  C12N15/74 ,  C12N1/21

    CPC分类号： C12P37/04 ,  C12N9/80 ,  C12N9/84

    摘要： The subject of the present invention is to provide a β-lactam acylase protein having high activity, a gene coding for said β-lactam acylase protein, a recombinant vector having said gene, a transformant containing said recombinant vector, and a method of producing a β-lactam antibiotic such as amoxycillin using said β-lactam acylase. A β-lactam acylase gene of Stenotrophomonas maltophilia was cloned, the DNA base sequence and the amino acid sequence expected therefrom was determined, and a Stenotrophomonas β-lactam acylase gene was obtained. This gene was found to code for a protein with a molecular weight of about 70 kDa and having β-lactam acylase activity, and could efficiently produce amoxycillin without being inhibited by phenylacetic acid, etc. Furthermore, by modification of the amino acid sequence, a protein which can more efficiently produce amoxycillin could be obtained.

    摘要翻译： 本发明的主题是提供具有高活性的β-内酰胺酰基转移酶蛋白，编码所述β-内酰胺酰基转移酶蛋白的基因，具有所述基因的重组载体，含有所述重组载体的转化体，以及制备 β-内酰胺抗生素如阿莫西林，使用所述β-内酰胺酰基转移酶。 克隆了嗜麦芽寡养单胞菌的β-内酰胺酰基转移酶基因，测定了DNA碱基序列和预期的氨基酸序列，得到了嗜麦芽寡养单胞菌β-内酰胺酰基转移酶基因。 发现该基因编码具有约70kDa分子量且具有β-内酰胺酰基转移酶活性的蛋白质，并且可以有效地产生阿莫西林而不被苯乙酸等抑制。此外，通过修饰氨基酸序列，a 可以获得更有效地产生阿莫西林的蛋白质。

    STABLE BIOCATALYSTS OF PENICILLIN ACYLASE AS GEL AGGREGATES AND THE PROCESS OF MANUFACTURE THEREOF
    7.
    发明申请
    STABLE BIOCATALYSTS OF PENICILLIN ACYLASE AS GEL AGGREGATES AND THE PROCESS OF MANUFACTURE THEREOF 失效
    扁桃酸酯酶作为凝胶聚集体的稳定生物活性物质及其制备方法

    公开(公告)号：US20110287509A1

    公开(公告)日：2011-11-24

    申请号：US12989716

    申请日：2008-11-14

    申请人： Anupama Datla ,  Williams Rajasekar Vyasarayani ,  Trupti Krishnakant Ashar ,  Kyslik Pavel ,  Becka Stanislav

    发明人： Anupama Datla ,  Williams Rajasekar Vyasarayani ,  Trupti Krishnakant Ashar ,  Kyslik Pavel ,  Becka Stanislav

    IPC分类号： C12N11/04

    CPC分类号： C12N11/04 ,  C12N9/84 ,  C12N9/96 ,  C12N11/06 ,  C12P35/02 ,  C12P37/04 ,  C12P37/06 ,  Y02P20/588

    摘要： The present invention provides a stable and viable biocatalyst with high activity and operational stability and method of immobilization thereof. The immobilization process is based on the principle of entrapment of partially purified enzyme precipitate which is simultaneously aggregated by cross linking agent like glutaraldehyde and further entrapped in a combination of natural polymer like gelatin and synthetic polymer like polyvinyl alcohol with effective gelation under mild conditions of temperature and pH, resulting in a stable biocatalyst. The enzymes immobilized by the above process include Penicillin acylase from rE. coli RE III (pKA18), Novel Penicillin acylase from Achromobacter sp (CCM4834) expressed in rE. coli BL21 (pK1P1) CCM 7394 and rE. coli RE III (pKX1P1).

    摘要翻译： 本发明提供了具有高活性和操作稳定性的稳定且可行的生物催化剂及其固定方法。 固定方法基于夹带部分纯化的酶沉淀物的原理，其同时由交联剂如戊二醛凝结，并进一步截留在天然聚合物如明胶和合成聚合物如聚乙烯醇的组合中，在温和的温度条件下有效凝胶化 和pH，导致稳定的生物催化剂。 通过上述方法固定的酶包括来自rE的青霉素酰基转移酶。 大肠杆菌RE III（pKA18），来自无色杆菌的新型青霉素酰基转移酶（CCM4834）以rE表达。 大肠杆菌BL21（pK1P1）CCM7394和rE。 大肠杆菌RE III（pKX1P1）。

    METHOD FOR PREPARING A BETA-LACTAM ANTIBIOTIC
    9.
    发明申请
    METHOD FOR PREPARING A BETA-LACTAM ANTIBIOTIC 无效
    制备BETA-LACTAM抗生素的方法

    公开(公告)号：US20020006642A1

    公开(公告)日：2002-01-17

    申请号：US09202799

    申请日：1999-05-03

    发明人： THOMAS VAN DER DOES ,  JAGDISH CHANDER KAPUR ,  ERIK DE VROOM

    IPC分类号： C12P037/04 ,  C12P021/04

    CPC分类号： C12P37/04 ,  C12P35/02 ,  C12P35/04 ,  C12P37/06

    摘要： The invention relates to a method for preparing a null-lactam antibiotic, wherein an N-substituted null-lactam, having general formula (I), wherein R0 is hydrogen or C1-3 alkoxy; Y is CH2, oxygen, sulfur, or an oxidized form of sulfur; Z is (a), (b), (c) or (d), wherein R1 is hydrogen, hydroxy, halogen, C1-3 alkoxy, optionally substituted, optionally containing one or more heteroatoms, saturated or unsaturated, branched or straight C1-5 alkyl, preferably methyl, optionally substituted, optionally containing one or more heteroatoms C5-8 cycloalkyl, optionally substituted aryl or heteroaryl, or optionally substituted benzyl; and X is (CH2)m-A-(CH2)n, wherein m and n are the same or different and are chosen from the group of integers 0, 1, 2, 3 or 4 and A is CHnullCH, CnullC, CHB, CnullO, optionally substituted nitrogen, oxygen, sulfur or an optionally oxidized form of sulfur, and B is hydrogen, halogen, hydroxy, C1-3 alkoxy, or optionally substituted methyl, or a salt thereof, is contacted with at least one dicarboxylate acylase, or a functional equivalent thereof, and reacted with a precursor for a side chain of the null-lactam antibiotic in the presence of at least one penicillin acylase, or a functional equivalent thereof.

    摘要翻译： 本发明涉及一种制备β-内酰胺抗生素的方法，其中具有通式（I）的N-取代的β-内酰胺，其中R 0是氢或C 1-3烷氧基; Y是CH 2，氧，硫或硫的氧化形式; Z是（a），（b），（c）或（d），其中R 1是氢，羟基，卤素，C 1-3烷氧基，任选地被取代，任选地含有一个或多个杂原子，饱和或不饱和的，支链或直链C1 -5烷基，优选甲基，任选地被取代，任选地含有一个或多个杂原子C5-8环烷基，任选取代的芳基或杂芳基，或任选取代的苄基; 并且X是（CH 2）mA-（CH 2）n，其中m和n相同或不同，并且选自整数0,1,2,3或4，A是CH = CH，C = C， CHB，C = O，任选取代的氮，氧，硫或任选氧化形式的硫，B是氢，卤素，羟基，C 1-3烷氧基或任选取代的甲基或其盐与至少 一种二羧酸酰基转移酶或其功能等同物，并且在至少一种青霉素酰基转移酶或其功能等同物的存在下与β-内酰胺抗生素的侧链的前体反应。

    Synthesis of beta-lactam antibiotics with immobilized penicillin amidase
    10.
    发明授权
    Synthesis of beta-lactam antibiotics with immobilized penicillin amidase 失效
    用固定青霉素酰胺酶合成β-内酰胺抗生素

    公开(公告)号：US06218138B1

    公开(公告)日：2001-04-17

    申请号：US09318732

    申请日：1999-05-26

    申请人： Ferhat Ilhan ,  Dieter Kraemer

    发明人： Ferhat Ilhan ,  Dieter Kraemer

    IPC分类号： C12P3704

    CPC分类号： C12P37/04 ,  C12P35/04

    摘要： Beta-lactam antibiotics are synthesized by reacting an amino-beta-lactam component with a corresponding amino-group-containing acylating side-chain component in the presence of penicillin amidase from E. coli covalently immobilized on support particles. The resulting beta-lactam antibiotic product is solubilized by adding an acid such as sulfuric acid to lower the pH to 1.0 at a temperature in the range of 0° C. to +5° C. The immobilized penicillin amidase is substantially inactivated by the acid. After separating the beta-lactam antibiotic product, the immobilized penicillin amidase is substantially reactivated for reuse in antibiotic synthesis by treatment with a buffer having about a neutral pH. Antibiotics that can be produced include ampicillin, amoxicillin, cephalexin, cefaclor and cefadroxil. Support particles that can be used include particles having a macroporous structure and a particle diameter of 10-1000 &mgr;m, particles having oxirane groups, particles made of a synthetic polymer and inorganic particles such as porous glass particles.

    摘要翻译： β-内酰胺抗生素通过氨基-β-内酰胺组分与相应的含氨基酰化侧链组分在共价固定在载体颗粒上的大肠杆菌的青霉素酰胺酶存在下反应而合成。 所得β-内酰胺抗生素产品通过加入酸如硫酸溶解，在0℃至+ 5℃的温度范围内将pH降至1.0。固定的青霉素酰胺酶基本上被酸灭活 。 在分离β-内酰胺抗生素产物之后，通过用具有约中性pH的缓冲液处理，固定化的青霉素酰胺酶基本上被再活化用于抗生素合成中的再利用。 可生产的抗生素包括氨苄青霉素，阿莫西林，头孢氨苄，头孢克洛和头孢羟氨苄。 可以使用的载体颗粒包括具有大孔结构，粒径为10-1000μm的颗粒，具有环氧乙烷基团的颗粒，由合成聚合物制成的颗粒和无机颗粒如多孔玻璃颗粒。