Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance

One major challenge in biofuel production, including biobutanol production, is the low tolerance of the microbial host towards increasing biofuel concentration during fermentation. Here, we have demonstrated that Escherichia coli 1-butanol tolerance can be greatly enhanced through random mutagenesis...

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Main Authors: Zhang, Hongfang, Chong, Huiqing, Ching, Chi Bun, Song, Hao, Jiang, Rongrong
Other Authors: School of Chemical and Biomedical Engineering
Format: Article
Language:English
Published: 2013
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Online Access:https://hdl.handle.net/10356/98865
http://hdl.handle.net/10220/12518
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-988652020-03-07T11:35:37Z Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance Zhang, Hongfang Chong, Huiqing Ching, Chi Bun Song, Hao Jiang, Rongrong School of Chemical and Biomedical Engineering DRNTU::Engineering::Chemical engineering::Biotechnology One major challenge in biofuel production, including biobutanol production, is the low tolerance of the microbial host towards increasing biofuel concentration during fermentation. Here, we have demonstrated that Escherichia coli 1-butanol tolerance can be greatly enhanced through random mutagenesis of global transcription factor cyclic AMP receptor protein (CRP). Four mutants (MT1–MT4) with elevated 1-butanol tolerance were isolated from error-prone PCR libraries through an enrichment screening. A DNA shuffling library was then constructed using MT1–MT4 as templates and one mutant (MT5) that exhibited the best tolerance ability among all variants was selected. In the presence of 0.8 % (v/v, 6.5 g/l) 1-butanol, the growth rate of MT5 was found to be 0.28 h−1 while that of wild type was 0.20 h−1. When 1-butanol concentration increased to 1.2 % (9.7 g/l), the growth rate of MT5 (0.18 h−1) became twice that of the wild type (0.09 h−1). Microbial adhesion to hydrocarbon test showed that cell surface of MT5 was less hydrophobic and its cell length became significantly longer in the presence of 1-butanol, as observed by scanning electron microscopy. Quantitative real-time reverse transcription PCR analysis revealed that several CRP regulated, 1-butanol stress response related genes (rpoH, ompF, sodA, manX, male, and marA) demonstrated differential expression in MT5 in the presence or absence of 1-butanol. In conclusion, direct manipulation of the transcript profile through engineering global transcription factor CRP can provide a useful tool in strain engineering. 2013-07-30T06:37:50Z 2019-12-06T20:00:37Z 2013-07-30T06:37:50Z 2019-12-06T20:00:37Z 2012 2012 Journal Article Zhang, H., Chong, H., Ching, C. B., Song, H.,& Jiang, R. (2012). Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance. Applied Microbiology and Biotechnology, 94(4), 1107-1117. https://hdl.handle.net/10356/98865 http://hdl.handle.net/10220/12518 10.1007/s00253-012-4012-5 en Applied microbiology and biotechnology
institution Nanyang Technological University
building NTU Library
country Singapore
collection DR-NTU
language English
topic DRNTU::Engineering::Chemical engineering::Biotechnology
spellingShingle DRNTU::Engineering::Chemical engineering::Biotechnology
Zhang, Hongfang
Chong, Huiqing
Ching, Chi Bun
Song, Hao
Jiang, Rongrong
Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance
description One major challenge in biofuel production, including biobutanol production, is the low tolerance of the microbial host towards increasing biofuel concentration during fermentation. Here, we have demonstrated that Escherichia coli 1-butanol tolerance can be greatly enhanced through random mutagenesis of global transcription factor cyclic AMP receptor protein (CRP). Four mutants (MT1–MT4) with elevated 1-butanol tolerance were isolated from error-prone PCR libraries through an enrichment screening. A DNA shuffling library was then constructed using MT1–MT4 as templates and one mutant (MT5) that exhibited the best tolerance ability among all variants was selected. In the presence of 0.8 % (v/v, 6.5 g/l) 1-butanol, the growth rate of MT5 was found to be 0.28 h−1 while that of wild type was 0.20 h−1. When 1-butanol concentration increased to 1.2 % (9.7 g/l), the growth rate of MT5 (0.18 h−1) became twice that of the wild type (0.09 h−1). Microbial adhesion to hydrocarbon test showed that cell surface of MT5 was less hydrophobic and its cell length became significantly longer in the presence of 1-butanol, as observed by scanning electron microscopy. Quantitative real-time reverse transcription PCR analysis revealed that several CRP regulated, 1-butanol stress response related genes (rpoH, ompF, sodA, manX, male, and marA) demonstrated differential expression in MT5 in the presence or absence of 1-butanol. In conclusion, direct manipulation of the transcript profile through engineering global transcription factor CRP can provide a useful tool in strain engineering.
author2 School of Chemical and Biomedical Engineering
author_facet School of Chemical and Biomedical Engineering
Zhang, Hongfang
Chong, Huiqing
Ching, Chi Bun
Song, Hao
Jiang, Rongrong
format Article
author Zhang, Hongfang
Chong, Huiqing
Ching, Chi Bun
Song, Hao
Jiang, Rongrong
author_sort Zhang, Hongfang
title Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance
title_short Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance
title_full Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance
title_fullStr Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance
title_full_unstemmed Engineering global transcription factor cyclic AMP receptor protein of Escherichia coli for improved 1-butanol tolerance
title_sort engineering global transcription factor cyclic amp receptor protein of escherichia coli for improved 1-butanol tolerance
publishDate 2013
url https://hdl.handle.net/10356/98865
http://hdl.handle.net/10220/12518
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