title: The expression of spinach glycolate oxidase (GO) in E. coli and the application of GO in the production of glyoxylic acid.
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 sulfatepolyacrylamide 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.
authors.affiliation: Dept. of Bioengineering, Beijing University of Chemical Technology, Beijing 100029, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Jin
authors.forename: Tianwei
authors.initials: T
authors.lastname: Tan
authors.forename: Hengliang
authors.initials: H
authors.lastname: Wang
authors.forename: Guofu
authors.initials: G
authors.lastname: Su
mesh_terms: D000429:Alcohol Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D005822:Genetic Vectors; D006038:Glyoxylates / Q000378:metabolism*; D007544:Isopropyl Thiogalactoside / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins / Q000096:biosynthesis; D018724:Spinacia oleracea / Q000201:enzymology* / Q000235:genetics
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D006038:Glyoxylates; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside; D000429:Alcohol Oxidoreductases; C020821:glycollate oxidase; C031150:glyoxylic acid