Improved production of the non-native cofactor F420 in Escherichia coli

Abstract The deazaflavin cofactor F420 is a low-potential, two-electron redox cofactor produced by some Archaea and Eubacteria that is involved in methanogenesis and methanotrophy, antibiotic biosynthesis, and xenobiotic metabolism. However, it is not produced by bacterial strains commonly used for...

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Autores principales:	Mihir V. Shah, Hadi Nazem-Bokaee, James Antoney, Suk Woo Kang, Colin J. Jackson, Colin Scott
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/32148b2317254743871f9cd92f54d262
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spelling	oai:doaj.org-article:32148b2317254743871f9cd92f54d2622021-11-08T10:52:46ZImproved production of the non-native cofactor F420 in Escherichia coli10.1038/s41598-021-01224-32045-2322https://doaj.org/article/32148b2317254743871f9cd92f54d2622021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01224-3https://doaj.org/toc/2045-2322Abstract The deazaflavin cofactor F420 is a low-potential, two-electron redox cofactor produced by some Archaea and Eubacteria that is involved in methanogenesis and methanotrophy, antibiotic biosynthesis, and xenobiotic metabolism. However, it is not produced by bacterial strains commonly used for industrial biocatalysis or recombinant protein production, such as Escherichia coli, limiting our ability to exploit it as an enzymatic cofactor and produce it in high yield. Here we have utilized a genome-scale metabolic model of E. coli and constraint-based metabolic modelling of cofactor F420 biosynthesis to optimize F420 production in E. coli. This analysis identified phospho-enol pyruvate (PEP) as a limiting precursor for F420 biosynthesis, explaining carbon source-dependent differences in productivity. PEP availability was improved by using gluconeogenic carbon sources and overexpression of PEP synthase. By improving PEP availability, we were able to achieve a ~ 40-fold increase in the space–time yield of F420 compared with the widely used recombinant Mycobacterium smegmatis expression system. This study establishes E. coli as an industrial F420-production system and will allow the recombinant in vivo use of F420-dependent enzymes for biocatalysis and protein engineering applications.Mihir V. ShahHadi Nazem-BokaeeJames AntoneySuk Woo KangColin J. JacksonColin ScottNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mihir V. Shah Hadi Nazem-Bokaee James Antoney Suk Woo Kang Colin J. Jackson Colin Scott Improved production of the non-native cofactor F420 in Escherichia coli
description	Abstract The deazaflavin cofactor F420 is a low-potential, two-electron redox cofactor produced by some Archaea and Eubacteria that is involved in methanogenesis and methanotrophy, antibiotic biosynthesis, and xenobiotic metabolism. However, it is not produced by bacterial strains commonly used for industrial biocatalysis or recombinant protein production, such as Escherichia coli, limiting our ability to exploit it as an enzymatic cofactor and produce it in high yield. Here we have utilized a genome-scale metabolic model of E. coli and constraint-based metabolic modelling of cofactor F420 biosynthesis to optimize F420 production in E. coli. This analysis identified phospho-enol pyruvate (PEP) as a limiting precursor for F420 biosynthesis, explaining carbon source-dependent differences in productivity. PEP availability was improved by using gluconeogenic carbon sources and overexpression of PEP synthase. By improving PEP availability, we were able to achieve a ~ 40-fold increase in the space–time yield of F420 compared with the widely used recombinant Mycobacterium smegmatis expression system. This study establishes E. coli as an industrial F420-production system and will allow the recombinant in vivo use of F420-dependent enzymes for biocatalysis and protein engineering applications.
format	article
author	Mihir V. Shah Hadi Nazem-Bokaee James Antoney Suk Woo Kang Colin J. Jackson Colin Scott
author_facet	Mihir V. Shah Hadi Nazem-Bokaee James Antoney Suk Woo Kang Colin J. Jackson Colin Scott
author_sort	Mihir V. Shah
title	Improved production of the non-native cofactor F420 in Escherichia coli
title_short	Improved production of the non-native cofactor F420 in Escherichia coli
title_full	Improved production of the non-native cofactor F420 in Escherichia coli
title_fullStr	Improved production of the non-native cofactor F420 in Escherichia coli
title_full_unstemmed	Improved production of the non-native cofactor F420 in Escherichia coli
title_sort	improved production of the non-native cofactor f420 in escherichia coli
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/32148b2317254743871f9cd92f54d262
work_keys_str_mv	AT mihirvshah improvedproductionofthenonnativecofactorf420inescherichiacoli AT hadinazembokaee improvedproductionofthenonnativecofactorf420inescherichiacoli AT jamesantoney improvedproductionofthenonnativecofactorf420inescherichiacoli AT sukwookang improvedproductionofthenonnativecofactorf420inescherichiacoli AT colinjjackson improvedproductionofthenonnativecofactorf420inescherichiacoli AT colinscott improvedproductionofthenonnativecofactorf420inescherichiacoli
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Improved production of the non-native cofactor F420 in Escherichia coli

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