Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.

Cofactor F(420) is a unique electron carrier in a number of microorganisms including Archaea and Mycobacteria. It has been shown that F(420) has a direct and important role in archaeal energy metabolism whereas the role of F(420) in mycobacterial metabolism has only begun to be uncovered in the last...

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Autores principales:	Ghader Bashiri, Aisyah M Rehan, David R Greenwood, James M J Dickson, Edward N Baker
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3506db2cc61840cc8e46470ba8a91f5a
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spelling	oai:doaj.org-article:3506db2cc61840cc8e46470ba8a91f5a2021-11-18T07:01:03ZMetabolic engineering of cofactor F420 production in Mycobacterium smegmatis.1932-620310.1371/journal.pone.0015803https://doaj.org/article/3506db2cc61840cc8e46470ba8a91f5a2010-12-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21209917/?tool=EBIhttps://doaj.org/toc/1932-6203Cofactor F(420) is a unique electron carrier in a number of microorganisms including Archaea and Mycobacteria. It has been shown that F(420) has a direct and important role in archaeal energy metabolism whereas the role of F(420) in mycobacterial metabolism has only begun to be uncovered in the last few years. It has been suggested that cofactor F(420) has a role in the pathogenesis of M. tuberculosis, the causative agent of tuberculosis. In the absence of a commercial source for F(420), M. smegmatis has previously been used to provide this cofactor for studies of the F(420)-dependent proteins from mycobacterial species. Three proteins have been shown to be involved in the F(420) biosynthesis in Mycobacteria and three other proteins have been demonstrated to be involved in F(420) metabolism. Here we report the over-expression of all of these proteins in M. smegmatis and testing of their importance for F(420) production. The results indicate that co-expression of the F(420) biosynthetic proteins can give rise to a much higher F(420) production level. This was achieved by designing and preparing a new T7 promoter-based co-expression shuttle vector. A combination of co-expression of the F(420) biosynthetic proteins and fine-tuning of the culture media has enabled us to achieve F(420) production levels of up to 10 times higher compared with the wild type M. smegmatis strain. The high levels of the F(420) produced in this study provide a suitable source of this cofactor for studies of F(420)-dependent proteins from other microorganisms and for possible biotechnological applications.Ghader BashiriAisyah M RehanDavid R GreenwoodJames M J DicksonEdward N BakerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 12, p e15803 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ghader Bashiri Aisyah M Rehan David R Greenwood James M J Dickson Edward N Baker Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.
description	Cofactor F(420) is a unique electron carrier in a number of microorganisms including Archaea and Mycobacteria. It has been shown that F(420) has a direct and important role in archaeal energy metabolism whereas the role of F(420) in mycobacterial metabolism has only begun to be uncovered in the last few years. It has been suggested that cofactor F(420) has a role in the pathogenesis of M. tuberculosis, the causative agent of tuberculosis. In the absence of a commercial source for F(420), M. smegmatis has previously been used to provide this cofactor for studies of the F(420)-dependent proteins from mycobacterial species. Three proteins have been shown to be involved in the F(420) biosynthesis in Mycobacteria and three other proteins have been demonstrated to be involved in F(420) metabolism. Here we report the over-expression of all of these proteins in M. smegmatis and testing of their importance for F(420) production. The results indicate that co-expression of the F(420) biosynthetic proteins can give rise to a much higher F(420) production level. This was achieved by designing and preparing a new T7 promoter-based co-expression shuttle vector. A combination of co-expression of the F(420) biosynthetic proteins and fine-tuning of the culture media has enabled us to achieve F(420) production levels of up to 10 times higher compared with the wild type M. smegmatis strain. The high levels of the F(420) produced in this study provide a suitable source of this cofactor for studies of F(420)-dependent proteins from other microorganisms and for possible biotechnological applications.
format	article
author	Ghader Bashiri Aisyah M Rehan David R Greenwood James M J Dickson Edward N Baker
author_facet	Ghader Bashiri Aisyah M Rehan David R Greenwood James M J Dickson Edward N Baker
author_sort	Ghader Bashiri
title	Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.
title_short	Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.
title_full	Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.
title_fullStr	Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.
title_full_unstemmed	Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.
title_sort	metabolic engineering of cofactor f420 production in mycobacterium smegmatis.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/3506db2cc61840cc8e46470ba8a91f5a
work_keys_str_mv	AT ghaderbashiri metabolicengineeringofcofactorf420productioninmycobacteriumsmegmatis AT aisyahmrehan metabolicengineeringofcofactorf420productioninmycobacteriumsmegmatis AT davidrgreenwood metabolicengineeringofcofactorf420productioninmycobacteriumsmegmatis AT jamesmjdickson metabolicengineeringofcofactorf420productioninmycobacteriumsmegmatis AT edwardnbaker metabolicengineeringofcofactorf420productioninmycobacteriumsmegmatis
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Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis.

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