Open AccessOverexpression of a hydrogenase gene in Clostridium paraputrificum to enhance hydrogen gas production
Author(s) -
Morimoto Kenji,
Kimura Tetsuya,
Sakka Kazuo,
Ohmiya Kunio
Publication year - 2005
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2005.04.014
Subject(s) - hydrogenase , escherichia coli , clostridium , biochemistry , gene , clostridium perfringens , biology , open reading frame , acetic acid , lactic acid , recombinant dna , chemistry , bacteria , peptide sequence , enzyme , genetics
A [Fe]‐hydrogenase gene ( hydA ) was cloned from Clostridium paraputrificum M‐21 in Escherichia coli using a conserved DNA sequence of clostridial hydrogenase genes amplified by PCR as the probe. The hydA gene consisted of an open reading frame of 1749 bp encoding 582 amino acids with an estimated molecular mass of 64,560 Da. It was ligated into a shuttle vector, pJIR751, originally constructed for Clostridium perfringens and E. coli , and expressed in C. paraputrificum . Hydrogen gas productivity of the recombinant increased up to 1.7‐fold compared with the wild‐type. In the recombinant, overexpression of hydA abolished lactic acid production and increased acetic acid production by over‐oxidation of NADH, which is required for reduction of pyruvic acid to lactic acid in the wild‐type.