Primary structure and catalytic mechanism of the epoxide hydrolase from Agrobacterium radiobacter AD1.

The Journal of Biological Chemistry
R RinkDick B Janssen

Abstract

The epoxide hydrolase gene from Agrobacterium radiobacter AD1, a bacterium that is able to grow on epichlorohydrin as the sole carbon source, was cloned by means of the polymerase chain reaction with two degenerate primers based on the N-terminal and C-terminal sequences of the enzyme. The epoxide hydrolase gene coded for a protein of 294 amino acids with a molecular mass of 34 kDa. An identical epoxide hydrolase gene was cloned from chromosomal DNA of the closely related strain A. radiobacter CFZ11. The recombinant epoxide hydrolase was expressed up to 40% of the total cellular protein content in Escherichia coli BL21(DE3) and the purified enzyme had a kcat of 21 s-1 with epichlorohydrin. Amino acid sequence similarity of the epoxide hydrolase with eukaryotic epoxide hydrolases, haloalkane dehalogenase from Xanthobacter autotrophicus GJ10, and bromoperoxidase A2 from Streptomyces aureofaciens indicated that it belonged to the alpha/beta-hydrolase fold family. This conclusion was supported by secondary structure predictions and analysis of the secondary structure with circular dichroism spectroscopy. The catalytic triad residues of epoxide hydrolase are proposed to be Asp107, His275, and Asp246. Replacement of these residues to...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Apr 1, 1992·Protein Engineering·D L OllisJ Schrag
Dec 18, 1991·European Journal of Biochemistry·M H JacobsD B Janssen
Feb 22, 1990·Nature·F K WinklerW Hunziker
Jan 1, 1990·Methods in Enzymology·F W StudierJ W Dubendorff
Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Jun 1, 1989·Archives of Biochemistry and Biophysics·C HassettC J Omiecinski
Jan 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·T A Kunkel
Mar 1, 1985·Applied and Environmental Microbiology·D B JanssenB Witholt
Aug 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·H R MorrisJ R Tippins
Jan 6, 1981·Biochemistry·S W Provencher, J Glöckner
Jan 1, 1995·DNA and Cell Biology·J K BeethamB D Hammock
Aug 1, 1994·The Plant Journal : for Cell and Molecular Biology·A StapletonW R Belknap
Aug 1, 1994·The Plant Journal : for Cell and Molecular Biology·T KiyosueK Shinozaki
Feb 1, 1994·Protein Engineering·C Geourjon, G Deléage
Mar 1, 1994·Chemical Research in Toxicology·G M Lacourciere, R N Armstrong
Aug 15, 1993·Archives of Biochemistry and Biophysics·J K BeethamB D Hammock
Mar 5, 1993·Journal of Molecular Biology·K M ArmstrongR L Baldwin
Dec 1, 1995·Protein Science : a Publication of the Protein Society·P K MehtaP Argos
Feb 23, 1996·The Journal of Biological Chemistry·M ArandF Oesch
Mar 18, 1996·Biochemical and Biophysical Research Communications·H Wojtasek, G D Prestwich
Dec 1, 1964·Analytical Biochemistry·R F ITZHAKI, D M GILL

❮ Previous
Next ❯

Citations

Jun 2, 2007·Applied Microbiology and Biotechnology·Jung-Hee WooSang-Jin Kim
Dec 17, 2008·Applied Microbiology and Biotechnology·Jung-Hee WooSang-Jin Kim
Sep 2, 2011·Applied Microbiology and Biotechnology·Vinayagam VasuSankaranarayanan Meenakshisundaram
Jan 23, 2008·Journal of Industrial Microbiology & Biotechnology·Guido MelzerPeter Götz
Mar 17, 2005·Biochimica Et Biophysica Acta·Lisa T Elfström, Mikael Widersten
Sep 10, 2003·Analytical Biochemistry·Kai DodererRolf D Schmid
Feb 13, 2001·Journal of Biotechnology·J E van Hylckama Vlieg, D B Janssen
Feb 19, 2002·Current Opinion in Biotechnology·A Steinreiber, K Faber
Dec 23, 1999·Current Opinion in Structural Biology·M Nardini, B W Dijkstra
Jul 17, 1998·Current Opinion in Chemical Biology·R J Kazlauskas, H K Weber
Feb 18, 1999·Current Opinion in Chemical Biology·R V Orru, K Faber
Feb 22, 2012·Journal of the American Chemical Society·Antonio C RuzziniLindsay D Eltis
Apr 7, 2012·Chemical Research in Toxicology·Rafael Gómez-BombarelliJulio Casado
Jul 17, 1999·European Journal of Biochemistry·C MorisseauJ C Baratti
Apr 7, 2006·Applied and Environmental Microbiology·Bert van LooDick B Janssen
Dec 10, 2013·ACS Synthetic Biology·Nagendra Prasad KurumbangJiri Damborsky
Jan 4, 2001·Drug Metabolism Reviews·R N Armstrong, C S Cassidy
Jan 7, 2004·Drug Metabolism Reviews·Michael ArandT Alwyn Jones
Mar 5, 1999·Drug Metabolism Reviews·R N Armstrong
Apr 15, 2008·FEBS Letters·Bert van LooDick B Janssen
Nov 4, 2015·ACS Catalysis·Beat A AmreinShina C L Kamerlin
May 3, 2014·Environmental Science & Technology·Pavel DvorakZbynek Prokop
May 18, 2004·Proteins·Sandra BarthJürgen Pleiss
Dec 14, 2005·Biochimica Et Biophysica Acta·Michael Kotik, Pavel Kyslík
Sep 20, 2011·Biochimica Et Biophysica Acta·Q Q HouC B Liu
Aug 23, 2011·Journal of Biotechnology·Michael KotikVladimír Křen

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.