Publisher DOI: 10.1002/elsc.201800118
Title: Improved electrocompetence and metabolic engineering of Clostridium pasteurianum reveals a new regulation pattern of glycerol fermentation
Language: English
Authors: Schmitz, Rebekka 
Sabra, Wael 
Arbter, Philipp 
Hong, Yaeseong 
Utesch, Tyll 
Zeng, An-Ping  
Issue Date: Jun-2019
Source: Engineering in Life Sciences 6 (19): 412-422 (2019-06)
Journal or Series Name: Engineering in life sciences 
Abstract (english): Clostridium pasteurianum produces industrially valuable chemicals such as n-butanol and 1,3-propanediol from fermentations of glycerol and glucose. Metabolic engineering for increased yields of selective compounds is not well established in this microorganism. In order to study carbon fluxes and to selectively increase butanol yields, we integrated the latest advances in genome editing to obtain an electrocompetent Clostridium pasteurianum strain for further engineering. Deletion of the glycerol dehydratase large subunit (dhaB) using an adapted S. pyogenes Type II CRISPR/Cas9 nickase system resulted in a 1,3-propanediol-deficient mutant producing butanol as the main product. Surprisingly, the mutant was able to grow on glycerol as the sole carbon source. In spite of reduced growth, butanol yields were highly increased. Metabolic flux analysis revealed an important role of the newly identified electron bifurcation pathway for crotonyl-CoA to butyryl-CoA conversion in the regulation of redox balance. Compared to the parental strain, the electron bifurcation pathway flux of the dhaB mutant increased from 8 to 46% of the overall flux from crotonyl-CoA to butyryl-CoA and butanol, indicating a new, 1,3-propanediol-independent pattern of glycerol fermentation in Clostridium pasteurianum.
ISSN: 1618-2863
Institute: Bioprozess- und Biosystemtechnik V-1 
Type: (wissenschaftlicher) Artikel
Funded by: A part of this work was financially supported by the European Union Seventh Framework Program (FP7/2007 2013) through the EuroBioRef project (grant agreement Nr. 241718).
Project: European Multilevel Integrated Biorefinery Design for Sustainable Biomass Processing 
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