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

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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.

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Authors and Affiliations

  1. Dept. of Bioengineering, Beijing University of Chemical Technology, 100029, Beijing, China

    Tianwei Tan

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  1. Jianfeng Jin
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  2. Tianwei Tan
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  3. Hengliang Wang
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  4. Guofu Su
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Correspondence to Tianwei Tan.

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Jin, J., Tan, T., Wang, H. et al. The expression of spinach glycolate oxidase (GO) in E. coli and the application of GO in the production of glyoxylic acid. Mol Biotechnol 25, 207–214 (2003). https://doi.org/10.1385/MB:25:3:207

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