O-405 バイオマス由来水素または合成ガスからの有用物質生産を目的とした好熱性微生物の遺伝子組換技術の開発(ガス・ケミカル,Oral Session4)

概要

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To develop a genetic transformation system for M. thermoacetica ATCC39073, the uracil auxotrophic strain dpyrF was constructed by disrupting pyrF for orotate monophosphate (OMP) decarboxylase. To test heterologous gene expression in dpyrF, the lactate dehydrogenase (LDH) gene (T-ldh) from Thermoanaerobacter pseudethanolicus ATCC33223 was electroporated into dpyrF with a promoter of the glyceraldehyde-3-phosphate dehydrogenase (G3PD) gene of M. thermoacetica ATCC39073. The resulting transformant successfully produced 6.8 mM of lactate from fructose. To develop a transformation system without auxotrophy, a thermostable kanamycin resistant gene (kan^r) derived from the plasmid that Streptococcus faecalis harbored was introduced into M. thermoacetica ATCC39073. By using kan^r with promoter of endogenous G3PD gene as a selection marker, we succeeded at the transformation. Using the developed system, we successfully constructed acetate-deficient, lactate producing mutant by deleting phosphotransacylase gene candidates and introducing T-ldh.
一般社団法人日本エネルギー学会の論文
2014-01-08