Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling
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 clone...
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fthighwire:oai:open-archive.highwire.org:proeng:17/2/133 2023-05-15T13:35:19+02:00 Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling Suen, Wen-Chen Zhang, Ningyan Xiao, Li Madison, Vincent Zaks, Aleksey 2004-02-01 00:00:00.0 text/html http://peds.oxfordjournals.org/cgi/content/short/17/2/133 https://doi.org/10.1093/protein/gzh017 en eng Oxford University Press http://peds.oxfordjournals.org/cgi/content/short/17/2/133 http://dx.doi.org/10.1093/protein/gzh017 Copyright (C) 2004, Oxford University Press ORIGINAL ARTICLES TEXT 2004 fthighwire https://doi.org/10.1093/protein/gzh017 2007-06-24T02:54:14Z 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 ( T m ) 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. Text Antarc* Antarctica HighWire Press (Stanford University) Protein Engineering Design and Selection 17 2 133 140 |
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HighWire Press (Stanford University) |
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ORIGINAL ARTICLES |
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ORIGINAL ARTICLES Suen, Wen-Chen Zhang, Ningyan Xiao, Li Madison, Vincent Zaks, Aleksey Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling |
topic_facet |
ORIGINAL ARTICLES |
description |
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 ( T m ) 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. |
format |
Text |
author |
Suen, Wen-Chen Zhang, Ningyan Xiao, Li Madison, Vincent Zaks, Aleksey |
author_facet |
Suen, Wen-Chen Zhang, Ningyan Xiao, Li Madison, Vincent Zaks, Aleksey |
author_sort |
Suen, Wen-Chen |
title |
Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling |
title_short |
Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling |
title_full |
Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling |
title_fullStr |
Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling |
title_full_unstemmed |
Improved activity and thermostability of Candida antarctica lipase B by DNA family shuffling |
title_sort |
improved activity and thermostability of candida antarctica lipase b by dna family shuffling |
publisher |
Oxford University Press |
publishDate |
2004 |
url |
http://peds.oxfordjournals.org/cgi/content/short/17/2/133 https://doi.org/10.1093/protein/gzh017 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
http://peds.oxfordjournals.org/cgi/content/short/17/2/133 http://dx.doi.org/10.1093/protein/gzh017 |
op_rights |
Copyright (C) 2004, Oxford University Press |
op_doi |
https://doi.org/10.1093/protein/gzh017 |
container_title |
Protein Engineering Design and Selection |
container_volume |
17 |
container_issue |
2 |
container_start_page |
133 |
op_container_end_page |
140 |
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1766064143411970048 |