| Title | Surface Display Expression of Bacillus licheniformis Lipase in Escherichia coli Using Lpp’OmpA Chimera |
|---|---|
| Author | Jae-Hyung Jo, Chan-Wook Han, Seung-Hwan Kim, Hyuk-Jin Kwon, and Hyune-Hwan Lee* |
| Address | 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 |
| Bibliography | Journal of Microbiology, 52(10),856-862, 2014, |
| DOI | 10.1007/s12275-014-4217-7 |
| Key Words | surface display, Bacillus lichenformis, lipase, Lpp’OmpA, Escherichia coli |
| 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. |