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Molecular Biotechnology

, Volume 25, Issue 3, pp 207–214 | Cite as

The expression of spinach glycolate oxidase (GO) in E. coli and the application of GO in the production of glyoxylic acid

  • Jianfeng Jin
  • Tianwei Tan
  • Hengliang Wang
  • Guofu Su
Research

Abstract

The complementary deoxyribonucleic acid (cDNA) coding spinach glycolate oxidase (GO) was amplified by reverse transcription polymerase chain reaction (RT-PCR), using the total ribonucleic acid (RNA) of spinach leaves as the template, and was cloned into cloning vector pMD18-T. After the DNA sequence was determined, the go gene was subcloned into Escherichia coli expression vectors. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that recombinant GO was expressed in E. coli BL21 (DE3)(pTIG-Trx-GO) and E. coli BL21 (DE3)(pET-22b(+)-GO). The result of the enzyme activity assay and the oxidation of glycolate to glyoxylate catalyzed by the whole cells of E. coli BL21 (DE3)(pET-22b(+)-GO) proved the feasibility of using E. coli cells to produce glyoxylic acid.

Index Entries

Glycolate oxidase glyoxylic acid cloning expression 

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Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  • Jianfeng Jin
  • Tianwei Tan
    • 1
  • Hengliang Wang
  • Guofu Su
  1. 1.Dept. of BioengineeringBeijing University of Chemical TechnologyBeijingChina

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