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  4. Improved electrocompetence and metabolic engineering of Clostridium pasteurianum reveals a new regulation pattern of glycerol fermentation
 
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Improved electrocompetence and metabolic engineering of Clostridium pasteurianum reveals a new regulation pattern of glycerol fermentation

Publikationstyp
Journal Article
Date Issued
2019-06
Sprache
English
Author(s)
Schmitz, Rebekka  
Sabra, Wael  
Arbter, Philipp  
Hong, Yaeseong  
Utesch, Tyll  
Zeng, An-Ping  orcid-logo
Institut
Bioprozess- und Biosystemtechnik V-1  
TORE-URI
http://hdl.handle.net/11420/2962
Journal
Engineering in life sciences  
Volume
19
Issue
6
Start Page
412
End Page
422
Citation
Engineering in Life Sciences 6 (19): 412-422 (2019-06)
Publisher DOI
10.1002/elsc.201800118
Scopus ID
2-s2.0-85057499257
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.
DDC Class
000: Allgemeines, Wissenschaft
Funding(s)
European Multilevel Integrated Biorefinery Design for Sustainable Biomass Processing  
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