Metabolic engineering of cofactor F420 production in mycobacterium smegmatis

Cofactor F420 is a unique electron carrier in a number of microorganisms including Archaea and Mycobacteria. It has been shown that F420 has a direct and important role in archaeal energy metabolism whereas the role of F420 in mycobacterial metabolism has only begun to be uncovered in the last few y...

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Main Authors:	Bashiri, Ghader, Mohamed Rehan, Aisyah, Greenwood, David R., Dickson, James M. J., Baker, Edward N.
Format:	Article
Language:	English
Published:	The Public Library of Science (PLoS) 2010
Subjects:	Q Science (General) QR Microbiology
Online Access:	http://irep.iium.edu.my/9925/ http://irep.iium.edu.my/9925/ http://irep.iium.edu.my/9925/ http://irep.iium.edu.my/9925/1/journal.pone.0015803.pdf

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spelling	iium-99252011-12-13T12:41:20Z http://irep.iium.edu.my/9925/ Metabolic engineering of cofactor F420 production in mycobacterium smegmatis Bashiri, Ghader Mohamed Rehan, Aisyah Greenwood, David R. Dickson, James M. J. Baker, Edward N. Q Science (General) QR Microbiology Cofactor F420 is a unique electron carrier in a number of microorganisms including Archaea and Mycobacteria. It has been shown that F420 has a direct and important role in archaeal energy metabolism whereas the role of F420 in mycobacterial metabolism has only begun to be uncovered in the last few years. It has been suggested that cofactor F420 has a role in the pathogenesis of M. tuberculosis, the causative agent of tuberculosis. In the absence of a commercial source for F420, M. smegmatis has previously been used to provide this cofactor for studies of the F420-dependent proteins from mycobacterial species. Three proteins have been shown to be involved in the F420 biosynthesis in Mycobacteria and three other proteins have been demonstrated to be involved in F420 metabolism. Here we report the over-expression of all of these proteins in M. smegmatis and testing of their importance for F420 production. The results indicate that co–expression of the F420 biosynthetic proteins can give rise to a much higher F420 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 F420 biosynthetic proteins and fine-tuning of the culture media has enabled us to achieve F420 production levels of up to 10 times higher compared with the wild type M. smegmatis strain. The high levels of the F420 produced in this study provide a suitable source of this cofactor for studies of F420-dependent proteins from other microorganisms and for possible biotechnological applications. The Public Library of Science (PLoS) 2010-12-29 Article PeerReviewed application/pdf en http://irep.iium.edu.my/9925/1/journal.pone.0015803.pdf Bashiri, Ghader and Mohamed Rehan, Aisyah and Greenwood, David R. and Dickson, James M. J. and Baker, Edward N. (2010) Metabolic engineering of cofactor F420 production in mycobacterium smegmatis. PLoS ONE, 5 (12). e15803. ISSN 1932-6203 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0015803 10.1371/journal.pone.0015803
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	Q Science (General) QR Microbiology
spellingShingle	Q Science (General) QR Microbiology Bashiri, Ghader Mohamed Rehan, Aisyah Greenwood, David R. Dickson, James M. J. Baker, Edward N. Metabolic engineering of cofactor F420 production in mycobacterium smegmatis
description	Cofactor F420 is a unique electron carrier in a number of microorganisms including Archaea and Mycobacteria. It has been shown that F420 has a direct and important role in archaeal energy metabolism whereas the role of F420 in mycobacterial metabolism has only begun to be uncovered in the last few years. It has been suggested that cofactor F420 has a role in the pathogenesis of M. tuberculosis, the causative agent of tuberculosis. In the absence of a commercial source for F420, M. smegmatis has previously been used to provide this cofactor for studies of the F420-dependent proteins from mycobacterial species. Three proteins have been shown to be involved in the F420 biosynthesis in Mycobacteria and three other proteins have been demonstrated to be involved in F420 metabolism. Here we report the over-expression of all of these proteins in M. smegmatis and testing of their importance for F420 production. The results indicate that co–expression of the F420 biosynthetic proteins can give rise to a much higher F420 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 F420 biosynthetic proteins and fine-tuning of the culture media has enabled us to achieve F420 production levels of up to 10 times higher compared with the wild type M. smegmatis strain. The high levels of the F420 produced in this study provide a suitable source of this cofactor for studies of F420-dependent proteins from other microorganisms and for possible biotechnological applications.
format	Article
author	Bashiri, Ghader Mohamed Rehan, Aisyah Greenwood, David R. Dickson, James M. J. Baker, Edward N.
author_facet	Bashiri, Ghader Mohamed Rehan, Aisyah Greenwood, David R. Dickson, James M. J. Baker, Edward N.
author_sort	Bashiri, Ghader
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	The Public Library of Science (PLoS)
publishDate	2010
url	http://irep.iium.edu.my/9925/ http://irep.iium.edu.my/9925/ http://irep.iium.edu.my/9925/ http://irep.iium.edu.my/9925/1/journal.pone.0015803.pdf
first_indexed	2023-09-18T20:19:42Z
last_indexed	2023-09-18T20:19:42Z
_version_	1777408024380440576

Metabolic engineering of cofactor F420 production in mycobacterium smegmatis

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