title: Surface display expression of Bacillus licheniformis lipase in Escherichia coli using Lpp'OmpA chimera.
abstract: The lipase from Bacillus licheniformis ATCC14580 was displayed on the cell surface of Escherichia coli using Lpp'OmpA as the anchoring protein. The expressed Lpp'OmpA-lipase fusion protein has a molecular weight of approximately 35 kDa, which was confirmed by SDS-PAGE and western blot analysis. The Lpp'OmpA-lipase fusion protein was located on the cell surface, as determined by immunofluorescence confocal microscopy and flow cytometry. The enzyme activity of the surface-displayed lipase showed clear halo around the colony. The cell surface-displayed lipase showed the highest activity of 248.12 ± 9.42 U/g (lyophilized cell) at the optimal temperature of 37°C and pH 8.0. The enzyme exhibited the highest activity toward the substrate p-nitrophenyl caprylate (C8). These results suggest that E. coli, which displayed the lipase on its surface, could be used as a whole cell biocatalyst.
authors.affiliation: Department of Bioscience and Biotechnology and Protein Research Center of GRRC, College of Natural Sciences, Hankuk University of Foreign Studies, Kyunggi-Do, 449-791, Republic of Korea.
authors.forename: Jae-Hyung
authors.initials: JH
authors.lastname: Jo
authors.forename: Chan-Wook
authors.initials: CW
authors.lastname: Han
authors.forename: Seung-Hwan
authors.initials: SH
authors.lastname: Kim
authors.forename: Hyuk-Jin
authors.initials: HJ
authors.lastname: Kwon
authors.forename: Hyune-Hwan
authors.initials: HH
authors.lastname: Lee
mesh_terms: D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D061505:Cell Surface Display Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D060847:Metabolic Engineering; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004800:Enzymes, Immobilized; D029968:Escherichia coli Proteins; D008074:Lipoproteins; C099932:Lpp protein, E coli; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C078167:OMPA outer membrane proteins; D008049:Lipase