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Open Access Research

Manipulating the sleeping beauty mutase operon for the production of 1-propanol in engineered Escherichia coli

Kajan Srirangan, Lamees Akawi, Xuejia Liu, Adam Westbrook, Eric JM Blondeel, Marc G Aucoin, Murray Moo-Young and C Perry Chou*

Author Affiliations

Department of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada

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Biotechnology for Biofuels 2013, 6:139  doi:10.1186/1754-6834-6-139

Published: 28 September 2013

Abstract

Background

While most resources in biofuels were directed towards implementing bioethanol programs, 1-propanol has recently received attention as a promising alternative biofuel. Nevertheless, no microorganism has been identified as a natural 1-propanol producer. In this study, we manipulated a novel metabolic pathway for the synthesis of 1-propanol in the genetically tractable bacterium Escherichia coli.

Results

E. coli strains capable of producing heterologous 1-propanol were engineered by extending the dissimilation of succinate via propionyl-CoA. This was accomplished by expressing a selection of key genes, i.e. (1) three native genes in the sleeping beauty mutase (Sbm) operon, i.e. sbm-ygfD-ygfG from E. coli, (2) the genes encoding bifunctional aldehyde/alcohol dehydrogenases (ADHs) from several microbial sources, and (3) the sucCD gene encoding succinyl-CoA synthetase from E. coli. Using the developed whole-cell biocatalyst under anaerobic conditions, production titers up to 150 mg/L of 1-propanol were obtained. In addition, several genetic and chemical effects on the production of 1-propanol were investigated, indicating that certain host-gene deletions could abolish 1-propanol production as well as that the expression of a putative protein kinase (encoded by ygfD/argK) was crucial for 1-propanol biosynthesis.

Conclusions

The study has provided a novel route for 1-propanol production in E. coli, which is subjected to further improvement by identifying limiting conversion steps, shifting major carbon flux to the productive pathway, and optimizing gene expression and culture conditions.

Keywords:
Bifunctional aldehyde/alcohol dehydrogenases; Cyanocobalamin; Metabolic engineering; Methylmalonyl-CoA mutases; Propanol; Propionate; Sleeping beauty mutase operon