An efficient bacterial surface display system based on a novel outer membrane anchoring element from the Escherichia coli protein YiaT

FEMS Microbiology Letters
Mee-Jung Han, Seung Hwan Lee

Abstract

In a bacterial surface display system, the display of a successful recombinant protein is highly dependent on the choice of anchoring motif. In this study, we developed an efficient Escherichia coli display system using novel anchoring motifs derived from the protein YiaT. To determine the best surface-anchoring motif, full-length YiaT and two of its C-terminal truncated forms, cut at the R181 and R232 sites, were evaluated. Two industrial enzymes, a lipase from Pseudomonas fluorescens SIK W1 and an α-amylase from Bacillus subtilis, were used as the target proteins for display. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot, immunofluorescence microscopy and whole-cell enzyme activity measurements confirmed the expression of the fusion proteins on the E. coli surface. Using YiaTR181 or YiaTR232 as the anchoring motif, the fusion proteins showed very high enzyme activities and did not exert any adverse effects on either cell growth or the outer membrane integrity. Additionally, these fusion proteins were suitable for displaying proteins of large molecular size in an active form. Compared with the previous anchoring motifs FadL and OprF, YiaTR181 and YiaTR232 had approximately 10-fold and 20-fold higher e...Continue Reading

References

Nov 1, 1993·Trends in Biotechnology·K Faber, M C Franssen
Feb 24, 2001·Nature·K M Koeller, C H Wong
Dec 26, 2001·Nature Biotechnology·Stephanie G BurtonJohn M Woodley
Jun 1, 2002·Journal of Biotechnology·Patrik SamuelsonStefan Ståhl
Dec 14, 2002·Trends in Biotechnology·Sang Yup LeeZhaohui Xu
Oct 11, 2003·Biotechnology Advances·I Benhar
Mar 24, 2004·Biotechnology and Applied Biochemistry·Henrik Wernérus, Stefan Ståhl
Jun 25, 2004·Nucleic Acids Research·Pantelis G BagosStavros J Hamodrakas
Sep 28, 2010·Biotechnology Letters·Kouichi Kuroda, Mitsuyoshi Ueda
Dec 15, 2010·Trends in Biotechnology·Edwin van BlooisMarco W Fraaije
Dec 22, 2010·Recent Patents on Biotechnology·Akihiko KondoChiaki Ogino
Apr 13, 2011·Microbial Cell Factories·Nguyen Thanh NhanGen Larsson

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Citations

Jan 23, 2018·Biotechnology Letters·H D Pardavé-AlejandreC R González-Bonilla
Dec 1, 2017·Journal of Industrial Microbiology & Biotechnology·Murali Kannan MaruthamuthuSoon Ho Hong
Apr 13, 2017·Frontiers in Microbiology·Abdullah F U H SaeedShihua Wang
Apr 24, 2020·Journal of Microbiology and Biotechnology·Mee Jung Han
Feb 26, 2021·Biotechnology Advances·Hao DongPing Wang
Jul 11, 2021·Trends in Biotechnology·Soo-Jin YeomChul-Ho Yun

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