Display of a thermostable lipase on the surface of a solvent-resistant bacterium, Pseudomonas putida GM730, and its applications in whole-cell biocatalysis.

BMC Biotechnology
Heung-Chae JungJae-Gu Pan

Abstract

Whole-cell biocatalysis in organic solvents has been widely applied to industrial bioprocesses. In two-phase water-solvent processes, substrate conversion yields and volumetric productivities can be limited by the toxicity of solvents to host cells and by the low mass transfer rates of the substrates from the solvent phase to the whole-cell biocatalysts in water. To solve the problem of solvent toxicity, we immobilized a thermostable lipase (TliA) from Pseudomonas fluorescens on the cell surface of a solvent-resistant bacterium, Pseudomonas putida GM730. Surface immobilization of enzymes eliminates the mass-transfer limitation imposed by the cell wall and membranes. TliA was successfully immobilized on the surface of P. putida cells using the ice-nucleation protein (INP) anchoring motif from Pseudomonas syrinage. The surface location was confirmed by flow cytometry, protease accessibility and whole-cell enzyme activity using a membrane-impermeable substrate. Three hundred and fifty units of whole-cell hydrolytic activity per gram dry cell mass were obtained when the enzyme was immobilized with a shorter INP anchoring motif (INPNC). The surface-immobilized TliA retained full enzyme activity in a two-phase water-isooctane reactio...Continue Reading

References

Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D H Figurski, D R Helinski
Sep 1, 1991·Agricultural and Biological Chemistry·G H ChungJ S Rhee
May 1, 1989·Trends in Biochemical Sciences·P Wolber, G Warren
Oct 12, 1996·Annals of the New York Academy of Sciences·H OginoH Ishikawa
Oct 23, 1998·Extremophiles : Life Under Extreme Conditions·S Isken, J A de Bont
Feb 13, 1999·Journal of Biotechnology·B Angelova, H P Schmauder
Feb 18, 1999·Current Opinion in Chemical Biology·Y L Khmelnitsky, J O Rich
May 14, 1999·Trends in Biotechnology·K N Timmis, D H Pieper
Jun 3, 2000·Nature Biotechnology·J S LeeC J Kim
Feb 24, 2001·Nature·A SchmidB Witholt
Aug 6, 2002·Research in Microbiology·Yogita Sardessai, Saroj Bhosle
Sep 5, 2002·Biotechnology and Bioengineering·Wilfred Chen, George Georgiou
Mar 15, 2003·Science·Hans E SchoemakerMarcel G Wubbolts
Dec 23, 2003·Applied Microbiology and Biotechnology·T MatsumotoA Kondo
Feb 18, 2004·Applied Microbiology and Biotechnology·R GuptaP Rathi
Mar 24, 2004·Biotechnology and Applied Biochemistry·Henrik Wernérus, Stefan Ståhl
Apr 6, 2004·Current Opinion in Chemical Biology·Nicholas J Turner
Apr 7, 2005·Current Opinion in Chemical Biology·Takeru IshigeSakayu Shimizu
Apr 23, 2005·Biochemical and Biophysical Research Communications·June-Hyung KimByung-Gee Kim
Oct 20, 2005·Journal of Bioscience and Bioengineering·Kyung-Su NaJunichi Kato

❮ Previous
Next ❯

Citations

Dec 25, 2007·Nature Reviews. Microbiology·Rotem SorekPhilip Hugenholtz
Aug 12, 2014·Applied Microbiology and Biotechnology·Jan SchüürmannJoachim Jose
Dec 11, 2007·Biotechnology Advances·M Saleem Zia-ul-Hassan
Jun 13, 2017·Journal of Molecular Microbiology and Biotechnology·Huayou ChenJinru Jia
Jul 26, 2008·Chirality·Azlina Harun KamaruddinHairul Nazirah Abdul Halim
Sep 8, 2012·Applied Biochemistry and Biotechnology·Saisubramanian Nagarajan
Sep 13, 2011·Microbial Biotechnology·Anika C Jahns, Bernd H A Rehm
Jul 1, 2014·Chembiochem : a European Journal of Chemical Biology·Robert J FloorDick B Janssen

❮ Previous
Next ❯

Datasets Mentioned

BETA
GM730

Methods Mentioned

BETA
flow cytometry

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.