Cloning and expression of the hydrogenase gene from Clostridium butyricum in Escherichia coli

Hydrogenase gene from Clostridium butyricum was cloned in Escherichia coli HK16 (Hyd−) using pBR322 and PstI. The plasmid, pCBH1, containing hydrogenase gene was 7.3 MDa and pCBH1 had 5 PstI-DNA fragments (3.9, 2.6, 0.7, 0.03–0.04, <0.02 MDa, respectively). The hydrogenase activity of HK16 (pCBH1) was about 3.1–3.5-times as high as those of the present strains, such as C. butyricum and E. coli C600 (Hyd+).