The expression of spinach glycolate oxidase (GO) in E. coli and the application of GO in the production of glyoxylic acid.
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.
Jin
Jianfeng
J
Dept. of Bioengineering, Beijing University of Chemical Technology, Beijing 100029, China.
Tan
Tianwei
T
Wang
Hengliang
H
Su
Guofu
G
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
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