Development of an in vivo methylation system for the solventogen Clostridium saccharobutylicum NCP 262 and analysis of two endonuclease mutants

Justyna Maria Lesiak; Wolfgang Liebl; Armin Ehrenreich

doi:10.1016/j.jbiotec.2014.07.005

Development of an in vivo methylation system for the solventogen Clostridium saccharobutylicum NCP 262 and analysis of two endonuclease mutants

J Biotechnol. 2014 Oct 20:188:97-9. doi: 10.1016/j.jbiotec.2014.07.005. Epub 2014 Aug 1.

Authors

Justyna Maria Lesiak¹, Wolfgang Liebl², Armin Ehrenreich³

Affiliations

¹ Lehrstuhl für Mikrobiologie, TU-München, Freising, Germany.
² Lehrstuhl für Mikrobiologie, TU-München, Freising, Germany. Electronic address: [email protected].
³ Lehrstuhl für Mikrobiologie, TU-München, Freising, Germany. Electronic address: [email protected].

PMID: 25087740
DOI: 10.1016/j.jbiotec.2014.07.005

Abstract

The genome of the biotechnologically important solventogenic Clostridium saccharobutylicum NCP 262 contains two operons coding for genes of presumed type I RM systems belonging to the families A and C. They represent a limiting factor for the development of transformation and conjugation protocols. We established an efficient triparental mating system to transfer DNA to C. saccharobutylicum by conjugation, which includes an in vivo methylation of the donor DNA. Furthermore we describe increased rates of conjugation in knock-out mutants of the restrictase subunits of both RM systems.

Keywords: ABE fermentation; C. saccharobutylicum NCP262; Conjugation; In vivo methylation; Type I restriction–modification system.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Clostridium / genetics
Clostridium / metabolism*
DNA Methylation*
DNA, Bacterial / metabolism
Endonucleases / genetics*
Mutation*

Substances

DNA, Bacterial
Endonucleases