Direct expression of active spinach glycolate oxidase in Escherichia coli.
Spinach glycolate oxidase (GAO) was expressed in Escherichia coli using the T7 RNA polymerase promotor. The enzyme accounts for approx. 1% of the soluble protein fraction and is expressed as a soluble and active enzyme. Comparison with GAO expressed in Saccharomyces cerevisiae (Macheroux, P., Massey, V., Thiele, D.J. and Volokita, M. (1991) Biochemistry 30, 4612-4619) showed that the GAO expressed in E. coli has identical physico-chemical features to the wild-type enzyme, but is expressed at a level approx. 15-fold higher than in the yeast system.
Macheroux
P
P
Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor.
Mulrooney
S B
SB
Williams
C H
CH
Massey
V
V
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D002845:Chromatography; D002850:Chromatography, Gel; D003001:Cloning, Molecular / Q000379:methods; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D006882:Hydroxyapatites; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D013604:T-Phages / Q000235:genetics
D006882:Hydroxyapatites; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D017886:Durapatite; D000429:Alcohol Oxidoreductases; C020821:glycollate oxidase