Overexpression of Escherichia coli genes encoding nucleoside phosphorylases in the pET/Bl21 (DE3) system yields active recombinant enzymes

被引：61

作者：

Esipov, RS ^{[1
]}

Gurevich, AI ^{[1
]}

Chuvikovsky, DV ^{[1
]}

Chupova, LA ^{[1
]}

Muravyova, TI ^{[1
]}

Miroshnikov, AI ^{[1
]}

机构：

[1] Russian Acad Sci, Shemyakin Ovchinnikov Inst Bioorgan Chem, Moscow 117997, Russia

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2002年 / 24卷 / 01期

关键词：

D O I：

10.1006/prep.2001.1524

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The Escherichia coli genes encoding purine nucleoside phosphorylase, uridine phosphorylase, and thymidine phosphorylase were cloned into pET plasmids to generate highly effective E. coli BL21(DE3) strains producing each of these enzymes. Optimum conditions for biosynthesis of each enzyme as a soluble protein with intact biological activity were found. The crude preparations are approximately 80% pure and can be used immediately for enzymatic transglycosylation. The enzyme preparations were purified to homogeneity by two steps including fractional precipitation with ammonium sulfate and subsequent chromatography on Sephadex G-100 and DEAE-Sephacel. (C) 2002 Elsevier Science (USA).

引用

页码：56 / 60

页数：5

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