Enhanced production of recombinant galactose oxidase from Fusarium graminearum in E. coli

被引：11

作者：

Choosri, Withu ^{[1
]}

Paukner, Regina ^{[1
]}

Wuehrer, Petra ^{[1
]}

Haltrich, Dietmar ^{[1
]}

Leitner, Christian ^{[1
]}

机构：

[1] Univ Nat Resources & Life Sci, BOKU, Food Biotechnol Lab, Dept Food Sci & Technol, A-1190 Vienna, Austria

来源：

WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY | 2011年 / 27卷 / 06期

关键词：

Fusarium graminearum; E; coli; Galactose oxidase; Overexpression; Fermentation; Affinity chromatography; DACTYLIUM-DENDROIDES; PURIFICATION; EXPRESSION; SEQUENCE; BINDING;

D O I：

10.1007/s11274-010-0585-2

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：

The gene gaoA encoding the copper-dependent enzyme galactose oxidase (GAO) from Fusarium graminearum PH-1 was cloned and successfully overexpressed in E. coli. Culture conditions for cultivations in shaken flasks were optimized, and optimal conditions were found to be double-strength LB medium, 0.5% lactose as inducer, and induction at the reduced temperature of 25A degrees C. When using these cultivation conditions similar to 24 mg of active GAO could be produced in shaken flasks per litre medium. Addition of copper to the fermentation medium decreased the enzyme production significantly. The His-tagged recombinant enzyme could be purified conveniently with a single affinity chromatography step. The purified enzyme showed a single band on SDS-PAGE with an apparent molecular mass of 66 kDa and had kinetic properties similar to those of the fungal wild-type enzyme.

引用

页码：1349 / 1353

页数：5

共 23 条

[1] PRODUCTION OF GALACTOSE-OXIDASE BY GIBBERELLA-FUJIKUROI [J].