M85 Repeated aspartate tags improve functional expression of Candida antarctica lipase B in recombinant Escherichia coli
Monday, April 27, 2015
Aventine Ballroom ABC/Grand Foyer, Ballroom Level
Sun-Ki Kim1, Won-Ki Min1, Ung Heo1, Yong-Cheol Park2, Hyung Ho Lee3, Seung Taeg Jeon3 and Jin-Ho Seo1, (1)Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul, (2)Bio and Fermentation Convergence Technology, Kookmin University, Seoul, (3)Bio and Nano Chemistry, Kookmin University, Seoul
 Escherichia coli is the best-established microbial host strain for production of proteins and chemicals, but has a weakness for not secreting high amounts of active heterologous proteins to the extracellular culture medium, of which origins belong to whether prokaryotes or eukaryotes. In this study, Candida antarctica lipase B (CalB), a popular eukaryotic enzyme which catalyzes a number of biochemical reactions and barely secreted extracellularly, was expressed functionally at a gram scale in culture medium by using a simple amino acid-tag system of E. coli. New fusion tag systems consisting of a pelB signal sequence and various anion amino acid tags facilitated both intracellular expression and extracellular secretion of CalB. Among them, the N-terminal five aspartate tag changed the quaternary structure of the dimeric CalB and allowed production of 1.9 g/L active CalB with 65 U/ml activity in culture medium, which exhibited the same enzymatic properties as the commercial CalB. This PelB anion amino acid tag-based expression system for CalB can be extended to production of other industrial proteins hardly expressed and exported from E. coli, thereby increasing target protein concentrations and minimizing purification steps.