Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration

Abstract Nitroaromatics are among the most important and commonly used chemicals but their production often suffers from multiple unsolved challenges. We have previously described the development of biocatalytic nitration processes driven by an engineered P450 TxtE fusion construct. Herein we report...

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Autores principales:	Ran Zuo, Yi Zhang, Chao Jiang, John C. Hackett, Rosemary Loria, Steven D. Bruner, Yousong Ding
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2d7e60a7e8a448e1b0cc0d1e7c0d1adf
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spelling	oai:doaj.org-article:2d7e60a7e8a448e1b0cc0d1e7c0d1adf2021-12-02T16:06:47ZEngineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration10.1038/s41598-017-00897-z2045-2322https://doaj.org/article/2d7e60a7e8a448e1b0cc0d1e7c0d1adf2017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00897-zhttps://doaj.org/toc/2045-2322Abstract Nitroaromatics are among the most important and commonly used chemicals but their production often suffers from multiple unsolved challenges. We have previously described the development of biocatalytic nitration processes driven by an engineered P450 TxtE fusion construct. Herein we report the creation of improved nitration biocatalysts through constructing and characterizing fusion proteins of TxtE with the reductase domain of CYP102A1 (P450BM3, BM3R). The majority of constructs contained variable linker length while one was rationally designed for optimizing protein-protein interactions. Detailed biochemical characterization identified multiple active chimeras that showed improved nitration activity, increased coupling efficiency and higher total turnover numbers compared with TxtE. Substrate promiscuity of the most active chimera was further assessed with a substrate library. Finally, a biocatalytic nitration process was developed to nitrate 4-Me-dl-Trp. The production of both 4-Me-5-NO2-l-Trp and 4-Me-7-NO2-l-Trp uncovered remarkable regio-promiscuity of nitration biocatalysts.Ran ZuoYi ZhangChao JiangJohn C. HackettRosemary LoriaSteven D. BrunerYousong DingNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ran Zuo Yi Zhang Chao Jiang John C. Hackett Rosemary Loria Steven D. Bruner Yousong Ding Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration
description	Abstract Nitroaromatics are among the most important and commonly used chemicals but their production often suffers from multiple unsolved challenges. We have previously described the development of biocatalytic nitration processes driven by an engineered P450 TxtE fusion construct. Herein we report the creation of improved nitration biocatalysts through constructing and characterizing fusion proteins of TxtE with the reductase domain of CYP102A1 (P450BM3, BM3R). The majority of constructs contained variable linker length while one was rationally designed for optimizing protein-protein interactions. Detailed biochemical characterization identified multiple active chimeras that showed improved nitration activity, increased coupling efficiency and higher total turnover numbers compared with TxtE. Substrate promiscuity of the most active chimera was further assessed with a substrate library. Finally, a biocatalytic nitration process was developed to nitrate 4-Me-dl-Trp. The production of both 4-Me-5-NO2-l-Trp and 4-Me-7-NO2-l-Trp uncovered remarkable regio-promiscuity of nitration biocatalysts.
format	article
author	Ran Zuo Yi Zhang Chao Jiang John C. Hackett Rosemary Loria Steven D. Bruner Yousong Ding
author_facet	Ran Zuo Yi Zhang Chao Jiang John C. Hackett Rosemary Loria Steven D. Bruner Yousong Ding
author_sort	Ran Zuo
title	Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration
title_short	Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration
title_full	Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration
title_fullStr	Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration
title_full_unstemmed	Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration
title_sort	engineered p450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/2d7e60a7e8a448e1b0cc0d1e7c0d1adf
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Engineered P450 biocatalysts show improved activity and regio-promiscuity in aromatic nitration

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