Korean Chemical Engineering Research

The Korean Institute of Chemical Engineers (한국화학공학회)
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Constitutive Expression of Lipase on the Cell Surface of Escherichia coli using OmpC Anchoring Motif

  • Lee, Seung Hwan (Department of Biotechnology & Bioengineering, Chonnam National University) ;
  • Lee, Sang Yup (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering (BK21 Program), Institute of BioCentury, Korea Advanced Institute of Science and Technology)
  • Received : 2020.01.17
  • Accepted : 2020.01.21
  • Published : 2020.05.01
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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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