Domestication of Clostridium sp. AWRP towards a Platform Strain for Syngas Fermentation

Abstract

Synthesis gas (Syngas) fermentation has been attracting attention since it can be prepared by gasification of various materials that can be directly utilized by microbes. Previously, we isolated a novel alcohologenic acetogen Clostridium sp. AWRP at Ansan. This bacterium is able to produce ethanol and 2,3-butanediol from either CO or CO2 plus H2, but as stated above, engineering this species had been hampered as well by recalcitrance against DNA transformation and non-optimized culture conditions. In this study, we introduce a recent progress in domestication of the AWRP strain: establishment of a genetic toolbox and improvement of culture conditions. In the first part, we discuss construction of Clostridium-E. coli shuttle plasmids and protocols of their transfer to the AWRP strain. The second part consists of modifications of culture medium, focusing on effects of trace elements and gas transfer.