Overexpression inEscherichia coliof a Recombinant ChimericRhodotorula gracilisd-Amino Acid Oxidase☆☆☆

摘要: Abstract This paper reports a novel expression system constructed to maximize the production in Escherichia coli of d -amino acid oxidase from yeast Rhodotorula gracilis (RgDAAO). We produced recombinant plasmid by insertion cDNA encoding for RgDAAO into multiple cloning site vector pT7.7 (pT7-DAAO), downstream T7 RNA polymerase binding site. The pT7-DAAO, which encodes fully active fusion protein with six additional residues at N-terminus DAAO, was used transform BL21(DE3) and BL21(DE3)pLysS E. cells. In latter host under optimal IPTG induction conditions, soluble chimeric DAAO expressed these cells up 930 U/g cell (and fermentation yield 2300 U/liter broth), specific activity 8.8 U/mg protein. represents ≈8% total content cell.