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Received January 17, 2020
Accepted January 21, 2020
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This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits
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Constitutive Expression of Lipase on the Cell Surface of Escherichia coli using OmpC Anchoring Motif
Department of Biotechnology & Bioengineering, Chonnam National University, 77 Yongbong-ro, Buk-gu, Gwangju, 61186, Korea 1Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering (BK21 Program), Institute of BioCentury, Korea Advanced Institute of Science and Technology, 291, Daehak-ro, Yeseong-gu, Daejeon, 34141, Korea
leesh@chonnam.ac.kr
Korean Chemical Engineering Research, May 2020, 58(2), 280-285(6), 10.9713/kcer.2020.58.2.280 Epub 4 May 2020
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Abstract
We have developed a constitutive display system of the Pseudomonas fluorescens SIK W1 TliA lipase on the cell surface of Escherichia coli using E. coli outer membrane protein C (OmpC) as an anchoring motif, which is an economical compared to induced system. For the constitutive expression of truncated OmpC-TliA fusion proteins, gntT104 promoter was employed. Cell growth was not affected by over expression of fusion protein during entire culture time, suggesting cell lysis was not a problem. The localization of truncated OmpC-TliA fusion protein on the cell surface was confirmed by immunofluorescence microscopy and measuring whole cell lipase activity. Constitutively displayed lipase was very stable, retaining activity enantioselectivity throughout the five repeated reactions. These results suggest that OmpC from E. coli be a useful anchoring motif for displaying enzymes on the cell surface without any inducers, and this stable surface display system can be employed for a broad range of biotechnological applications.
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References
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Smith MR, Khera E, Wen F, Ing. Eng. Chem. Res., 54(16), 4021 (2015)
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Shimazu M, Nguyen A, Mulchandani A, Chen W, Biotechnol. Prog., 19(5), 1612 (2003)
Lee SH, Choi JI, Park SJ, Lee SY, Park BC, Appl. Environ. Microbiol., 70, 5074 (2004)
Han ZL, Han SY, Zheng SP, Lin Y, Appl. Microbiol. Biotechnol., 85(1), 117 (2009)
Sambrook J, Russell DW, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2001).
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Xu Z, Lee SY, Appl. Environ. Microbiol., 65, 5142 (1999)
Park SJ, Park JP, Lee SY, Fems Microbiol. Lett, 214, 217 (2002)
Lee SH, Choi JI, Han MJ, Choi JH, Lee SY, Biotechnol. Bioeng., 90(2), 223 (2005)
Jeanteur D, Lakey JH, Pattus F, Mol. Microbiol., 5, 2153 (1991)
Matsumoto T, Ito M, Fukuda H, Kondo A, Appl. Microbiol. Biotechnol., 64(4), 481 (2004)
Lee H, Park SJ, Han MJ, Eom GT, Choi MJ, Kim SH, Oh YH, Song BK, Lee SH, Biotechnol. Lett., 35(10), 1677 (2013)
