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 245...
Published in: | Protein Engineering Design and Selection |
---|---|
Main Authors: | , , , , |
Format: | Text |
Language: | English |
Published: |
Oxford University Press
2004
|
Subjects: | |
Online Access: | http://peds.oxfordjournals.org/cgi/content/short/gzh017v1 https://doi.org/10.1093/protein/gzh017 |
id |
fthighwire:oai:open-archive.highwire.org:proeng:gzh017v1 |
---|---|
record_format |
openpolar |
spelling |
fthighwire:oai:open-archive.highwire.org:proeng:gzh017v1 2023-05-15T13:51:54+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 2004-01-12 06:35:28.0 text/html http://peds.oxfordjournals.org/cgi/content/short/gzh017v1 https://doi.org/10.1093/protein/gzh017 en eng Oxford University Press http://peds.oxfordjournals.org/cgi/content/short/gzh017v1 http://dx.doi.org/10.1093/protein/gzh017 Copyright (C) 2004, Oxford University Press Article TEXT 2004 fthighwire https://doi.org/10.1093/protein/gzh017 2007-06-24T10:40:53Z 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 Candida antarctica ATCC 32657 and a 13-fold higher activity than the most active parent derived from Crytococcus 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 Candida antarctica ATCC 32657 by 11-fold and 6.4°C, 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 |
institution |
Open Polar |
collection |
HighWire Press (Stanford University) |
op_collection_id |
fthighwire |
language |
English |
topic |
Article |
spellingShingle |
Article 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 |
Article |
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 Candida antarctica ATCC 32657 and a 13-fold higher activity than the most active parent derived from Crytococcus 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 Candida antarctica ATCC 32657 by 11-fold and 6.4°C, 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/gzh017v1 https://doi.org/10.1093/protein/gzh017 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
http://peds.oxfordjournals.org/cgi/content/short/gzh017v1 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 |
_version_ |
1766255936398163968 |