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

被引:2
作者
Lee, Seung Hwan [1 ]
Lee, Sang Yup [2 ]
机构
[1] Chonnam Natl Univ, Dept Biotechnol & Bioengn, 77 Yongbong Ro, Gwangju 61186, South Korea
[2] Korea Adv Inst Sci & Technol, Inst BioCentury, Metab & Biomol Engn Natl Res Lab, Dept Chem & Biomol Engn,BK21 Program, 291 Daehak Ro, Daejeon 34141, South Korea
来源
KOREAN CHEMICAL ENGINEERING RESEARCH | 2020年 / 58卷 / 02期
关键词
Cell surface display; Lipase; Constitutive expression; Outer membrane protein; Escherichia coli; DISPLAY; ENZYME; BIOCATALYSTS; SEQUENCE;
D O I
10.9713/kcer.2020.58.2.280
中图分类号
TQ [化学工业];
学科分类号
0817 ;
摘要
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.
引用
收藏
页码:280 / 285
页数:6
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