Surface Display Expression of Bacillus licheniformis Lipase in Escherichia coli Using Lpp’OmpA Chimera

Jae-Hyung Jo; Chan-Wook Han; Seung-Hwan Kim; Hyuk-Jin Kwon; Hyune-Hwan Lee

doi:10.1007/s12275-014-4217-7

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Surface Display Expression of Bacillus licheniformis Lipase in Escherichia coli Using Lpp’OmpA Chimera: Jae-Hyung Jo , Chan-Wook Han , Seung-Hwan Kim , Hyuk-Jin Kwon , Hyune-Hwan Lee; Journal of Microbiology 2014;52(10):856-862
DOI: https://doi.org/10.1007/s12275-014-4217-7
Published online: August 27, 2014

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 KoreaDepartment 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

Corresponding author: Hyune-Hwan Lee , Tel: +82-31-330-4280,

Received: 8 April 2014 • Revised: 11 July 2014 • Accepted: 15 July 2014

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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.

Keywords: surface display; Bacillus lichenformis; lipase; Lpp’OmpA; Escherichia coli

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Surface Display Expression of Bacillus licheniformis Lipase in Escherichia coli Using Lpp’OmpA Chimera

J. Microbiol. 2014;52(10):856-862. Published online August 27, 2014

DOI: https://doi.org/10.1007/s12275-014-4217-7

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