Skip Navigation


PEDS Advance Access originally published online on January 12, 2004
Protein Engineering Design and Selection 2004 17(2):133-140; doi:10.1093/protein/gzh017
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
17/2/133    most recent
gzh017v1
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 ISI Web of Science
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 arrow Search for citing articles in:
ISI Web of Science (24)
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Suen, W.-C.
Right arrow Articles by Zaks, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Suen, W.-C.
Right arrow Articles by Zaks, A.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2004 Oxford University Press

Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling

Wen-Chen Suen1,2, Ningyan Zhang1, Li Xiao3, Vincent Madison3 and Aleksey Zaks1

1Biotransformations Group and 3Structural Chemistry, Schering-Plough Research Institute, U-13-3000, 1011 Morris Avenue, Union, NJ 07083, USA

2 To whom correspondence should be addressed. e-mail: wen-chen.suen{at}spcorp.com

DNA family shuffling was used to create chimeric lipase B proteins with improved activity toward the hydrolysis of diethyl 3-(3',4'-dichlorophenyl)glutarate (DDG). Three homologous lipases from Candida antarctica ATCC 32657, Hyphozyma sp. CBS 648.91 and Crytococcus tsukubaensis ATCC 24555 were cloned and shuffled to generate a diverse gene library. A high-throughput screening assay was developed and used successfully to identify chimeric lipase B proteins having a 20-fold higher activity toward DDG than lipase B from C.antarctica ATCC 32657 and a 13-fold higher activity than the most active parent derived from C.tsukubaensis ATCC 24555. In addition, the stability characteristics of several highly active chimeric proteins were also improved as a result of family shuffling. For example, the half-life at 45°C and melting point (Tm) of one chimera exceeded those of lipase B from C.antarctica ATCC 32657 by 11-fold and 6.4°C, respectively, which closely approached the stability characteristics of the most thermostable parent derived from Hyphozyma sp. CBS 648.91.

Received September 2, 2003; revised November 17, 2003; accepted December 10, 2003 Edited by Andrew Griffiths


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


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
A. Suemori and M. Iwakura
A Systematic and Comprehensive Combinatorial Approach to Simultaneously Improve the Activity, Reaction Specificity, and Thermal Stability of p-Hydroxybenzoate Hydroxylase
J. Biol. Chem., July 6, 2007; 282(27): 19969 - 19978.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
R. Fujii, Y. Nakagawa, J. Hiratake, A. Sogabe, and K. Sakata
Directed evolution of Pseudomonas aeruginosa lipase for improved amide-hydrolyzing activity
Protein Eng. Des. Sel., February 1, 2005; 18(2): 93 - 101.
[Abstract] [Full Text] [PDF]


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.