Constitutive Expression of Lipase on the Cell Surface of Escherichia coli using OmpC Anchoring Motif

Seung Hwan Lee; Sang Yup Lee

Korean Chemical Engineering Research, Vol.58, No.2, 280-285, April, 2020

DOI10.9713/kcer.2020.58.2.280 Export Citation

Constitutive Expression of Lipase on the Cell Surface of Escherichia coli using OmpC Anchoring Motif

Seung Hwan Lee^†, and Sang Yup Lee¹

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

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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.

Keywords:Cell surface display;Lipase;Constitutive expression;Outer membrane protein;Escherichia coli

[References]

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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)

[1] Jaeger KE, Dijkstra BW, Reetz MT, Annu. Rev. Microbiol., 53, 315 (1999)

[2] Jaeger KE, Eggert T, Curr. Opin. Biotechnol., 13(4), 390 (2002)

[3] Reetz MT, Curr. Opin. Chem. Biol., 6(2), 145 (2002)

[4] Stergiou YS, Foukis A, Filippou M, Koukouritaki M, et al., Biotechnol. Advan., 31(8), 1846 (2013)

[5] Gotor-Fernandez V, Brieva R, Gotor V, J. Mol. Catal. B-Enzym., 40(3-4), 111 (2006)

[6] Rodrigues RC, Virgen-Ortiz JJ, dos Santos JCS, Alcantar AR, Barbosa O, Ortiz C, Fernandez-Lafuente R, Biotechnol. Adv., 37(5), 746 (2019)

[7] Van Beilen JB, Li Z, Curr. Opin. Biotechnol., 13(4), 338 (2002)

[8] Rodrigues RC, Ortiz C, Berenguer-Murcia A, Torres R, Fernandez-Lafuente R, Chem. Soc. Rev., 42(15), 6290 (2013)

[9] Linqiu C, Carrier-bound immobilized enzymes: principles, application and design, WILEY-VCH, Weinheim, Germany (2005).

[10] Krajewska B, Enzyme Microb. Technol., 35(2-3), 126 (2004)

[11] Shiraga S, Kawakami M, Ishiguro M, Ueda M, Appl. Environ. Microbiol., 71(8), 4335 (2005)

[12] Georgiou G, Stathopoulos C, Daugherty PS, Nayak AR, Iverson BL, Curtiss RI, Nat. Biotechnol., 15, 29 (1997)

[13] Lee SY, Choi JH, Xu J, Trends Biotechnol., 21, 45 (2003)

[14] Smith MR, Khera E, Wen F, Ing. Eng. Chem. Res., 54(16), 4021 (2015)

[15] Liu Z, Ho SH, Hasunuma T, Chang JS, Ren NQ, Kondo A, Bioresour. Technol., 215, 324 (2016)

[16] Matsumoto T, Fukuda H, Ueda M, Tanaka A, Kondo A, Appl. Environ. Microbiol., 68(9), 4517 (2002)

[17] Shimazu M, Nguyen A, Mulchandani A, Chen W, Biotechnol. Prog., 19(5), 1612 (2003)

[18] Lee SH, Choi JI, Park SJ, Lee SY, Park BC, Appl. Environ. Microbiol., 70, 5074 (2004)

[19] Han ZL, Han SY, Zheng SP, Lin Y, Appl. Microbiol. Biotechnol., 85(1), 117 (2009)

[20] Sambrook J, Russell DW, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2001).

[21] Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V, Riley M, Colladovides J, Glasner JD, Rode CK, Mayhew GF, Gregor J, Davis NW, Kirkpatrick HA, Goeden MA, Rose DJ, Mau B, Shao Y, Science, 277(5331), 1453 (1997)

[22] Xu Z, Lee SY, Appl. Environ. Microbiol., 65, 5142 (1999)

[23] Park SJ, Park JP, Lee SY, Fems Microbiol. Lett, 214, 217 (2002)

[24] Lee SH, Choi JI, Han MJ, Choi JH, Lee SY, Biotechnol. Bioeng., 90(2), 223 (2005)

[25] Jeanteur D, Lakey JH, Pattus F, Mol. Microbiol., 5, 2153 (1991)

[26] Matsumoto T, Ito M, Fukuda H, Kondo A, Appl. Microbiol. Biotechnol., 64(4), 481 (2004)

[27] Lee H, Park SJ, Han MJ, Eom GT, Choi MJ, Kim SH, Oh YH, Song BK, Lee SH, Biotechnol. Lett., 35(10), 1677 (2013)