Skip Navigation



PEDS Advance Access published online on January 28, 2009
Protein Engineering Design and Selection, doi:10.1093/protein/gzn081
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
22/4/257    most recent
gzn081v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Nayeem, A.
Right arrow Articles by Basch, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nayeem, A.
Right arrow Articles by Basch, J.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2009. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Engineering enzymes for improved catalytic efficiency: a computational study of site mutagenesis in epothilone-B hydroxylase

Akbar Nayeem1,3, Shu-Jen Chiang2, Suo-Win Liu2, Yuhua Sun2, Li You2 and Jonathan Basch2,3

1Computer Aided Drug Design, Division of Molecular Biosciences, Bristol-Myers Squibb Company, PO Box 5400, Princeton, NJ 08543-5400 2Technical Operations, Bristol-Myers Squibb Company, 6000 Thompson Road, East Syracuse, NY 13057-5050, USA

3 To whom correspondence should be addressed. E-mail: akbar.nayeem{at}bms.com (A.K.); jonathan.basch{at}bms.com (J.B.)

Epothilone F, 21-hydroxyl-epothilone B, is an intermediate in the synthesis of BMS-310705, an antitumor compound that has been evaluated in Phase I clinical trials. A bioconversion process utilizing the Gram-positive bacterium Amycolatopsis orientalis was used to prepare epothilone F from epothilone B. In order to improve the yield of epothilone F, a mutagenesis program was performed with the goal of engineering the epothilone-B hydroxylase (EBH) enzyme to improve the yield of epothilone F through oxidative biotransformation. The mutations in EBH increased the yield of epothilone F from 21% in the recombinant expression system to higher than 80% utilizing the best EBH mutants. The studies described here show how a homology model of EBH was used to obtain an understanding of the possible mechanism that led to improved yield of epothilone F in the mutated enzymes. A novel aspect of this study is that it provides some insight into how mutations distant from the binding site can affect enzyme activity.

Keywords: biocatalysis/cytochrome P450/epothilone/homology modeling/mutagenesis

Received February 5, 2008; revised October 21, 2008; accepted December 1, 2008.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.