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[大肠杆菌K-12尿苷磷酸化酶的蛋白质工程。I.产气克雷伯菌和鼠伤寒沙门氏菌尿苷磷酸化酶基因在大肠杆菌中的克隆与表达]

[Protein engineering of uridine phosphorylase from Escherichia coli K-12. I. Cloning and expression of uridine phosphorylase genes from Klebsiella aerogenes and Salmonella typhimurium in E. coli].

作者信息

Veĭko V P, Chebotaev D V, Ovcharova I V, Gul'ko L B

机构信息

State Research Institute of Genetics and Selection of Industrial Microorganisms, Moscow, Russia.

出版信息

Bioorg Khim. 1998 May;24(5):381-7.

PMID:9661793
Abstract

Genes of uridine phosphorylases (udp) from Klebsiella aerogenes and Salmonella typhimurium were cloned and expressed. Highly effective producer strains of the corresponding proteins were constructed. Enzymic properties of the UPases obtained were studied and compared with those from the Escherichia coli enzyme. Mutant forms of UPase from E. coli (D5E, D5N, D5A) were prepared by site-directed mutagenesis techniques. It was shown that the Asp5 residue plays an insignificant role in the formation of the active form of the protein.

摘要

对产气克雷伯菌和鼠伤寒沙门氏菌的尿苷磷酸化酶(udp)基因进行了克隆和表达。构建了相应蛋白质的高效生产菌株。研究了所得尿苷磷酸化酶的酶学性质,并与大肠杆菌来源的酶进行了比较。通过定点诱变技术制备了大肠杆菌尿苷磷酸化酶的突变形式(D5E、D5N、D5A)。结果表明,Asp5残基在蛋白质活性形式的形成中起的作用不显著。

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