title: Polycationic amino acid tags enhance soluble expression of Candida antarctica lipase B in recombinant Escherichia coli.
abstract: Lipase (EC 3.1.1.3) is a popular enzyme used as an ingredient in detergents and biocatalyst in many biochemical reactions. Lipase is usually expressed in Escherichia coli as an inactive inclusion body and at a low level. In this study, Candida antarctica lipase B (CalB) was fused with various polycationic amino acid tags and expressed in E. coli in order to increase a soluble expression level. By induction with 1.0 mM IPTG, the authentic and fused CalBs were expressed at 27-56% of total protein. The 10-arginine and 10-lysine tags fused at the C-terminal of CalB significantly increased the solubility of CalB by five- to ninefold, relative to the case of the authentic CalB expressed in a recombinant E. coli Origami 2™ (DE3) strain. Among a series of the C-terminal poly-arginine tags, the recombinant CalB combined with the 10-arginine tag (CalB-R10) possessed the highest lipase specific activity of 9.5 ± 0.03 U/mg protein, corresponding to a fourfold enhancement compared with the authentic CalB.
authors.affiliation: Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Hyun-Jung
authors.initials: HJ
authors.lastname: Jung
authors.forename: Sun-Ki
authors.initials: SK
authors.lastname: Kim
authors.forename: Won-Ki
authors.initials: WK
authors.lastname: Min
authors.forename: Sung-Suk
authors.initials: SS
authors.lastname: Lee
authors.forename: Kyungmoon
authors.initials: K
authors.lastname: Park
authors.forename: Yong-Cheol
authors.initials: YC
authors.lastname: Park
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Seo
mesh_terms: D000596:Amino Acids / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005656:Fungal Proteins; D002479:Inclusion Bodies / Q000201:enzymology*; D008049:Lipase / Q000096:biosynthesis* / Q000235:genetics; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D011073:Polyamines / Q000737:chemistry; D000071228:Polyelectrolytes; D011107:Polylysine / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012995:Solubility
chemical_list: D000596:Amino Acids; D005656:Fungal Proteins; D008049:Lipase; D010455:Peptides; D011073:Polyamines; D000071228:Polyelectrolytes; D011107:Polylysine; D011993:Recombinant Fusion Proteins; C015462:polyarginine; C431349:lipase B, Candida antarctica