Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene

Biological treatment is a cost-effective and environmentally friendly technique for treating toluene. However, this process is limited by the type and quantity of toluene degraders that can be applied to different environments and methods. It is a feasible solution to constructing new genetic engine...

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Autores principales:	Bo Wang, Feng Gao, Jing Xu, Jianjie Gao, Zhenjun Li, Lijuan Wang, Fujian Zhang, Yu Wang, Yongsheng Tian, Rihe Peng, Quanhong Yao
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2021
Materias:	toluene artificial gene cluster gene optimization bioremediation benzene successive hydroxylation Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/c10ce77f59e34d16b2409522e60ef139
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spelling	oai:doaj.org-article:c10ce77f59e34d16b2409522e60ef1392021-11-17T14:21:55ZOptimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene1310-28181314-353010.1080/13102818.2021.1996267https://doaj.org/article/c10ce77f59e34d16b2409522e60ef1392021-01-01T00:00:00Zhttp://dx.doi.org/10.1080/13102818.2021.1996267https://doaj.org/toc/1310-2818https://doaj.org/toc/1314-3530Biological treatment is a cost-effective and environmentally friendly technique for treating toluene. However, this process is limited by the type and quantity of toluene degraders that can be applied to different environments and methods. It is a feasible solution to constructing new genetic engineering organisms for toluene degradation through biotechnology. To facilitate genetic manipulation and improve gene expression, the toluene/o-xylene monooxygenase locus from Pseudomonas stutzeri was reconstructed and artificially synthesized. All six genes were optimized and driven with a monocistronic transcriptional pattern by T7 promoter and terminator to be expressed at a high level. The engineered bacteria can hydroxylate toluene to a mixture of o-, m- and p-cresol, which are then further oxidized to 3- and 4-methylcatechol. Benzene can also be oxidized to phenol, catechol and pyrogallol successively. The negative effects of toluene can be removed by the engineered bacteria via proteomics analysis. Overall, the artificial gene clusters established in this study can be used to construct different toluene and benzene degradation organisms for bioremediation. Supplemental data for this article is available online at https://doi.org/10.1080/13102818.2021.1996267 .Bo WangFeng GaoJing XuJianjie GaoZhenjun LiLijuan WangFujian ZhangYu WangYongsheng TianRihe PengQuanhong YaoTaylor & Francis Grouparticletolueneartificial gene clustergene optimizationbioremediationbenzenesuccessive hydroxylationBiotechnologyTP248.13-248.65ENBiotechnology & Biotechnological Equipment, Vol 35, Iss 1, Pp 1632-1642 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	toluene artificial gene cluster gene optimization bioremediation benzene successive hydroxylation Biotechnology TP248.13-248.65
spellingShingle	toluene artificial gene cluster gene optimization bioremediation benzene successive hydroxylation Biotechnology TP248.13-248.65 Bo Wang Feng Gao Jing Xu Jianjie Gao Zhenjun Li Lijuan Wang Fujian Zhang Yu Wang Yongsheng Tian Rihe Peng Quanhong Yao Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene
description	Biological treatment is a cost-effective and environmentally friendly technique for treating toluene. However, this process is limited by the type and quantity of toluene degraders that can be applied to different environments and methods. It is a feasible solution to constructing new genetic engineering organisms for toluene degradation through biotechnology. To facilitate genetic manipulation and improve gene expression, the toluene/o-xylene monooxygenase locus from Pseudomonas stutzeri was reconstructed and artificially synthesized. All six genes were optimized and driven with a monocistronic transcriptional pattern by T7 promoter and terminator to be expressed at a high level. The engineered bacteria can hydroxylate toluene to a mixture of o-, m- and p-cresol, which are then further oxidized to 3- and 4-methylcatechol. Benzene can also be oxidized to phenol, catechol and pyrogallol successively. The negative effects of toluene can be removed by the engineered bacteria via proteomics analysis. Overall, the artificial gene clusters established in this study can be used to construct different toluene and benzene degradation organisms for bioremediation. Supplemental data for this article is available online at https://doi.org/10.1080/13102818.2021.1996267 .
format	article
author	Bo Wang Feng Gao Jing Xu Jianjie Gao Zhenjun Li Lijuan Wang Fujian Zhang Yu Wang Yongsheng Tian Rihe Peng Quanhong Yao
author_facet	Bo Wang Feng Gao Jing Xu Jianjie Gao Zhenjun Li Lijuan Wang Fujian Zhang Yu Wang Yongsheng Tian Rihe Peng Quanhong Yao
author_sort	Bo Wang
title	Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene
title_short	Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene
title_full	Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene
title_fullStr	Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene
title_full_unstemmed	Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene
title_sort	optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from pseudomonas stutzeri in escherichia coli and its successive hydroxylation of toluene and benzene
publisher	Taylor & Francis Group
publishDate	2021
url	https://doaj.org/article/c10ce77f59e34d16b2409522e60ef139
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Optimization, reconstruction and heterologous expression of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri in Escherichia coli and its successive hydroxylation of toluene and benzene

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