초록 |
[FeFe]-hydrogenases in clostridia catalyze the evolution of molecular hydrogen from two protons (H+) and electrons. Overexpression of the hydA gene, encoding [FeFe]-hydrogenase, was used as a strategy to enhance hydrogen production in this study.A 4.3-kb DNA region of Clostridium tyrobutyricum, containing the hydA gene, was isolated using a genome walking method. A 2.3-kb DNA region containing the ORF and a possible promoter region was amplified and subcloned into the pJIR418 shuttle vector to construct the recombinant plasmid pJIR418-hydA. For homologous overexpression of hydA, the plasmid was then successfully transferred into C. tyrobutyricum with an electroporation protocol newly developed in this study. As a result, the hydrogen yield of the recombinantstrain in batch culture was1.5-fold higher than that of the wild-type strain. |