Functional expression of lipase A from Candida antarctica in Escherichia coli—A prerequisite for high-throughput screening and directed evolution

• Published in: Journal of molecular catalysis. B, Enzymatic Vol. 45; no. 1; pp. 62 - 67
• Main Authors: Pfeffer, Jan, Rusnak, Monika, Hansen, Carl-Erik, Rhlid, Rachid Bel, Schmid, Rolf D., Maurer, Steffen C.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.04.2007; Elsevier Science
• Subjects: Biological and medical sciences; Biotechnology; Candida antarctica; Chaperone co-expression; Escherichia coli; Functional expression; Fundamental and applied biological sciences. Psychology; Lipase; Methods. Procedures. Technologies; Protein engineering; Candida antarctica; Lipase; Functional expression; Escherichia coli; Chaperone co-expression; High throughput screening; Directed molecular evolution; candida antarctica; Enzyme; Chaperone; Coexpression; Triacylglycerol lipase; Esterases; Gene expression; Carboxylic ester hydrolases; Fungi; Bacteria; Hydrolases; Fungi Imperfecti; Recombinant protein; Enterobacteriaceae; Thallophyta

We report for the first time the functional and heterologous expression of lipase A from Candida antarctica (CalA) in the cytoplasm of Escherichia coli Origami™ B cells. Expression under control of the lac promoter in the pUC18 vector yielded 0.7 U mg −1 lipase activity, whereas expression of a thio...