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Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum

Schwarz, Katrin; Grosse-Honebrink, Alexander; Derecka, Kamila; Rotta, Carlo; Zhang, Ying; Minton, Nigel P.

Authors

Katrin Schwarz Katrin.Schwarz@nottingham.ac.ukacs@nottingham.ac.uk

Alexander Grosse-Honebrink

Kamila Derecka

Carlo Rotta

Ying Zhang

Nigel P. Minton nigel.minton@nottingham.ac.uk



Abstract

Declining fossil fuel reserves, coupled with environmental concerns over their continued extraction and exploitation have led to strenuous efforts to identify renewable routes to energy and fuels. One attractive option is to convert glycerol, a by-product of the biodiesel industry, into n-butanol, an industrially important chemical and potential liquid transportation fuel, using Clostridium pasteurianum. Under certain growth conditions this Clostridium species has been shown to predominantly produce n-butanol, together with ethanol and 1, 3-propanediol, when grown on glycerol. Further increases in the yields of n-butanol produced by C. pasteurianum could be accomplished through rational metabolic engineering of the strain. Accordingly, in the current report we have developed and exemplified a robust tool kit for the metabolic engineering of C. pasteurianum and used the system to make, for the first time, in-frame deletion mutants of pivotal genes involved in solvent production, namely hydA (hydrogenase), rex (Redox response regulator) and dhaBCE (glycerol dehydratase). We were, for the first time in C. pasteurianum, able to eliminate 1, 3-propanediol synthesis and demonstrate its production was essential for growth on glycerol as a carbon source. Inactivation of both rex and hydA resulted in increased n-butanol titres, representing the first steps towards improving the utilisation of C. pasteurianum as a chassis for the industrial production of this important chemical.

Journal Article Type Article
Journal Metabolic Engineering
Print ISSN 1096-7176
Electronic ISSN 1096-7184
Publisher Elsevier
Peer Reviewed Peer Reviewed
APA6 Citation Schwarz, K., Grosse-Honebrink, A., Derecka, K., Rotta, C., Zhang, Y., & Minton, N. P. (in press). Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum. Metabolic Engineering, https://doi.org/10.1016/j.ymben.2017.01.009
DOI https://doi.org/10.1016/j.ymben.2017.01.009
Keywords Clostridium pasteurianum; rex; hydA; dhaBCE; butanol; 1; 3-propanediol (PDO)
Publisher URL http://www.sciencedirect.com/science/article/pii/S1096717616302397
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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