Display of bacterial lipase on the Escherichia coli cell surface by using FadL as an anchoring motif and use of the enzyme in enantioselective biocatalysis

Applied and Environmental Microbiology
Seung Hwan LeeByoung Chul Park

Abstract

We have developed a novel cell surface display system by employing FadL as an anchoring motif, which is an outer membrane protein involved in long-chain fatty acid transport in Escherichia coli. A thermostable Bacillus sp. strain TG43 lipase (44.5 kDa) could be successfully displayed on the cell surface of E. coli in an active form by C-terminal deletion-fusion of lipase at the ninth external loop of FadL. The localization of the truncated FadL-lipase fusion protein on the cell surface was confirmed by confocal microscopy and Western blot analysis. Lipase activity was mainly detected with whole cells, but not with the culture supernatant, suggesting that cell lysis was not a problem. The activity of cell surface-displayed lipase was examined at different temperatures and pHs and was found to be the highest at 50 degrees C and pH 9 to 10. Cell surface-displayed lipase was quite stable, even at 60 and 70 degrees C, and retained over 90% of the full activity after incubation at 50 degrees C for a week. As a potential application, cell surface-displayed lipase was used as a whole-cell catalyst for kinetic resolution of racemic methyl mandelate. In 36 h of reaction, (S)-mandelic acid could be produced with the enantiomeric excess of...Continue Reading

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Citations

Aug 3, 2010·Applied Microbiology and Biotechnology·Wen-Shan LiuYun-Jun Yan
Aug 2, 2011·Biotechnology Letters·Heyun ZhaoYunjun Yan
Dec 2, 2009·Applied and Environmental Microbiology·Jong Hwan BaekSang Yup Lee
Mar 25, 2014·Applied and Environmental Microbiology·Bai-fen SongLi-jie Tang
Aug 17, 2014·Journal of Biotechnology·Iasson E P TozakidisJoachim Jose
Jan 5, 2010·Journal of Biotechnology·Mee-Jung HanWon Hee Han
Oct 29, 2009·Journal of Biotechnology·Tsutomu TanakaAkihiko Kondo
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Aug 29, 2014·The Journal of Microbiology·Jae-Hyung JoHyune-Hwan Lee
Apr 24, 2020·Journal of Microbiology and Biotechnology·Mee Jung Han
Sep 11, 2010·Journal of Agricultural and Food Chemistry·Wenshan LiuYunjun Yan

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