title: Expression of a lipase on the cell-surface of Escherichia coli using the OmpW anchoring motif and its application to enantioselective reactions.
abstract: Microbial-surface display is the expression of proteins or peptides on the surface of cells by fusing an appropriate protein as an anchoring motif. Here, the outer membrane protein W (OmpW) was selected as a fusion partner for functional expression of Pseudomonas fluorescence SIK W1 lipase (TliA) on the cell-surface of Escherichia coli. Localization of the truncated OmpW-TliA fusion protein on the cell-surface was confirmed by immunoblotting and functional assay of lipase activity. Enantioselective hydrolysis of rac-phenylethyl butanoate by the displayed lipase resulted in optically active (R)-phenyl ethanol with 96% enantiomeric excess and 44% of conversion in 5 days. Thus, a small outer membrane protein OmpW, is a useful anchoring motif for displaying an active enzyme of ~50 kDa on the cell-surface and the surface-displayed lipase can be employed as an enantioselective biocatalyst in organic synthesis.
authors.affiliation: Division of Drug Discovery Research, Korea Research Institute of Chemical Technology, P.O. Box 107, 141 Gajeong-ro, Yuseong-gu, Daejeon, 305-600, Republic of Korea, leeh@krict.re.kr.
authors.forename: Hyuk
authors.initials: H
authors.lastname: Lee
authors.forename: Si Jae
authors.initials: SJ
authors.lastname: Park
authors.forename: Mee-Jung
authors.initials: MJ
authors.lastname: Han
authors.forename: Gyeong Tae
authors.initials: GT
authors.lastname: Eom
authors.forename: Min-Jung
authors.initials: MJ
authors.lastname: Choi
authors.forename: Seong Ho
authors.initials: SH
authors.lastname: Kim
authors.forename: Young Hoon
authors.initials: YH
authors.lastname: Oh
authors.forename: Bong Keun
authors.initials: BK
authors.lastname: Song
authors.forename: Seung Hwan
authors.initials: SH
authors.lastname: Lee
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002087:Butyrates / Q000378:metabolism; D061505:Cell Surface Display Techniques / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D015151:Immunoblotting; D008049:Lipase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008970:Molecular Weight; D010626:Phenylethyl Alcohol / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D001425:Bacterial Outer Membrane Proteins; D002087:Butyrates; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; C064035:ompW protein, E coli; D008049:Lipase; D010626:Phenylethyl Alcohol