Hydrogenase Expression for Hydrogen Production and Characterization of. β-N-Acetylglucosaminidase of Clostridium paraputrificum M-21

摘要

Ahydrogenase gene (hyd) was cloned from Clostridium paraputrificum M-21 in Escherichia coli using a probe of a conserved DNA sequence of clostridia hydrogenase genes amplified by PCR. The cloned hyd was ligated into a shuttle vector between E. coli and C. paraputrificum M-21 and was expressed in C. paraputrificumM-21. The hyd consists of an open reading frame of 1749-bp, encoding 582 amino acids with a MW of 64560. The hydrogen gas productivity of the transformant was increased up to 1.8-fold. ?-N-acetylglucosaminidases gene (nag3A) was cloned from C. paraputrificum M-21 in Escherichia coli. The nag3A consists of an ORF of 1,239-bp, encoding 413 amino acids with a MW of 45,531. Nag3A is a single domain enzyme belonging to a family 3 catalytic domain of glycoside hydrolases. Nag3A was purified from a recombinant E. coli and characterized.