Fujii Yasuyo, Yamasaki Yoshiaki, Matsumoto Masahiro, Nishida Hiroyuki, Hada Megumi, Ohkubo Katsutoshi
Institute of Advanced Energy, Kyoto University, Uji 611-0011, Japan.
Biosci Biotechnol Biochem. 2004 Aug;68(8):1722-7. doi: 10.1271/bbb.68.1722.
A unique variant of glutathione independent formaldehyde dehydrogenase of Pseudomonas putida was obtained by random mutagenesis using the PCR-reaction. This YM042 mutant, S318G, was a cold-adapted formaldehyde dehyrogenase. The activity at 29 degrees C of the variant was 1.7-fold higher than that of the wild type. The K(m) values of the mutant at 37 degrees C were 0.40 mM for NAD(+) and 2.5 mM for formaldehyde, while those of the wild-type were 0.18 mM for NAD(+) and 2.1 mM for formaldehyde. The catalytic efficiency for formaldehyde was about 1.5-fold greater in the mutant than in the wild-type enzyme. The optimum pHs and temperatures of the mutant and the wild-type enzyme were 7.5, and 8.0 and 37 degrees C, and 47 degrees C, respectively. The thermal stability of the mutant was lower than that of the wild type.
