Structural characterization of the organic solvent-stable cholesterol oxidase from Chromobacterium sp. DS-1

Journal of Structural Biology
Martin SagermannNoriyuki Doukyu

Abstract

Cholesterol oxidase is of significant commercial interest as it is widely used as a biosensor for the detection of cholesterol in clinical samples, blood serum and food. Increased stability of this enzyme with regards to temperature and different solvent conditions are of great importance to the reliability and versatility of its applications. We here report the crystal structure of the cholesterol oxidase of Chromobacterium sp. DS-1 (CHOLOX). In contrast to other previously characterized cholesterol oxidases, this enzyme retains high activity in organic solvents and detergents at temperatures above 85 degrees C despite its mesophilic origin. With the availability of one other homologous oxidase of known three-dimensional structure, a detailed comparison of its sequence and structure was performed to elucidate the mechanisms of stabilization. In contrast to factors that typically contribute to the stability of thermophilic proteins, the structure of CHOLOX exhibits a larger overall cavity volume, less charged residues and less salt bridge interactions. Moreover, the vast majority of residue substitutions were found on or near the protein's solvent exposed surface. We propose that the engineering of enhanced stability may also b...Continue Reading

References

May 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·D W HeinzB W Matthews
Sep 1, 1994·Protein Science : a Publication of the Protein Society·L LinN R Kallenbach
Jun 11, 1993·Science·M BlaberB W Matthews
Aug 1, 1995·Protein Science : a Publication of the Protein Society·V Z SpassovR Ladenstein
Feb 1, 1996·Journal of Molecular Graphics·W HumphreyK Schulten
Nov 1, 1995·Journal of Molecular Recognition : JMR·N PattabiramanP J Fleming
May 13, 1997·Proceedings of the National Academy of Sciences of the United States of America·P D AdamsA T Brünger
Jan 1, 1997·Folding & Design·G Vogt, P Argos
Oct 3, 1998·Acta Crystallographica. Section D, Biological Crystallography·A T BrüngerG L Warren
Dec 23, 1998·Protein Engineering·A Karshikoff, R Ladenstein
Oct 27, 1999·Proceedings of the National Academy of Sciences of the United States of America·B SpillerR C Stevens
Mar 21, 2000·FEBS Letters·S Chakravarty, R Varadarajan
Apr 25, 2000·Protein Engineering·S KumarR Nussinov
May 24, 2000·The Journal of Steroid Biochemistry and Molecular Biology·J MacLachlanC J Brooks
Aug 23, 2001·Proceedings of the National Academy of Sciences of the United States of America·N A BakerJ A McCammon
Oct 2, 2001·Cellular and Molecular Life Sciences : CMLS·S Kumar, R Nussinov
Jul 16, 2003·Journal of Molecular Biology·Angela R CriswellGeorge N Phillips
Sep 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·UNKNOWN Collaborative Computational Project, Number 4
Mar 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·G J Kleywegt, T A Jones
May 1, 1997·Acta Crystallographica. Section D, Biological Crystallography·G N MurshudovE J Dodson
Nov 1, 2005·Current Opinion in Microbiology·Ksenia Egorova, Garabed Antranikian
Jun 23, 2007·Nucleic Acids Research·K G TinaN Srinivasan
Jun 27, 2007·Chembiochem : a European Journal of Chemical Biology·Dieter Perl, Franz Xaver Schmid
Dec 21, 2007·Acta Crystallographica. Section D, Biological Crystallography·Stefano Trapani, Jorge Navaza
Sep 27, 2008·Bioinformatics·L HolmA Schenkel
Nov 19, 2008·Applied Microbiology and Biotechnology·Noriyuki DoukyuMartin Sagermann
Apr 3, 2009·Biosensors & Bioelectronics·Sunil K AryaBansi D Malhotra
Jun 6, 2009·Applied Microbiology and Biotechnology·Noriyuki Doukyu

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.