Construction of butanol-producing pathway from Clostridium saccharobutylicum in Escherichia coli JM109 (DE3) and its fermentation

Abstract:

Abstract:［Objective］Several key genes (thlA，bcs-operon/crt-bcd1-etfB2-fixB2-hbd and adhE) in butanol pathway from Clostridium saccharobutylicum DSM13864 were cloned，and a butanol-producing Escherichia coli strain was successfully constructed．［Methods］Using genome of Clostridium saccharobutylicum DSM13864 as template，the key genes in butanol synthesis pathway were amplified，the recombinant plasmids pETDuet-bcs and pRSFDuet-thlA-adhE were constructed． Then the resultant plasmids were transformed into E.coli JM109(DE3) to obtain E． coli BUT1 for butanol production，under the semi-anaerobic condition．Effects of different mediums on butanol production were studied． ［Results］The recombinant E.coli was capable of producing butanol (25.4 mg/L) under semi-anaerobic fermentation．After optimization on the fermentation medium，butanol titer reached 34.1 mg/L．［Conclusion］Butanol production by recombinant E． coli harboring exogenous butanol-producing pathway from Clostridium saccharobutylicum provides a feasible solution to overcome the hurdles in traditional butanol production approach by Clostridia.

Key words:Keywords: Clostridium saccharobutylicum，Escherichia coli JM109(DE3)，butanol，semi-anaerobic fermentation