Construction of hybrid bacterial deacetoxycephalosporin C synthases (expandases) by in vivo homeologous recombination

被引：6
作者：
Adrio, JL
Hintermann, GA
Demain, AL
Piret, JM [1 ]
机构：
[1] Northeastern Univ, Dept Biol, Boston, MA 02115 USA
[2] MIT, Dept Biol, Cambridge, MA 02139 USA
来源：
ENZYME AND MICROBIAL TECHNOLOGY | 2002年 / 31卷 / 07期
关键词：
deacetoxycephalosporin C synthase; expandase; hybrid enzymes; Streptomyces clavuligerus; Nocardia lactamdurans;
D O I：
10.1016/S0141-0229(02)00179-5
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Homeologous recombination (recombination between partially homeologous DNA sequences) was used to produce novel functional deacetoxycephalosporin C synthase (expandase) enzymes in vivo which are hybrids of the Streptomyces clavuligerus and Nocardia lactamdurans enzymes. DNA sequencing of hybrids obtained in E. coli showed that recombination had occurred at several locations within conserved sequences as short as 2 bp. Recombination events obtained in a Streptomyces background resulted in expandases with altered activity on penicillin G as determined by bioassay and HPLC. (C) 2002 Elsevier Science Inc. All rights reserved.
引用
收藏
页码：932 / 940
页数：9
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