Biochemical characterization of NADH:FMN oxidoreductase HcbA3 from Nocardioides sp. PD653 in catalyzing aerobic HCB dechlorination

Autor:	Ryota Kataoka, Hiromasa Kiyota, Kazuhiro Takagi, Koji Ito
Rok vydání:	2020
Předmět:	chemistry.chemical_classification Enzyme chemistry Biochemistry Health Toxicology and Mutagenesis Insect Science Flavin reductase Halogenation Flavin group Enzyme kinetics Oxidative phosphorylation Reductase Dehalogenase
Zdroj:	Journal of Pesticide Science. 45:125-131
ISSN:	1349-0923 1348-589X
Popis:	Nocardioides sp. PD653 genes hcbA1, hcbA2, and hcbA3 encode enzymes that catalyze the oxidative dehalogenation of hexachlorobenzene (HCB), which is one of the most recalcitrant persistent organic pollutants (POPs). In this study, HcbA1, HcbA2, and HcbA3 were heterologously expressed and characterized. Among the flavin species tested, HcbA3 showed the highest affinity for FMN with a Kd value of 0.75±0.17 µM. Kinetic assays revealed that HcbA3 followed a ping-pong bi-bi mechanism for the reduction of flavins. The Km for NADH and FMN was 51.66±11.58 µM and 4.43±0.69 µM, respectively. For both NADH and FMN, the Vmax and kcat were 2.21±0.86 µM and 66.74±5.91 sec-1, respectively. We also successfully reconstituted the oxidative dehalogenase reaction in vitro, which consisted of HcbA1, HcbA3, FMN, and NADH, suggesting that HcbA3 may be the partner reductase component for HcbA1 in Nocardioides sp. PD653.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::490e91d6e6851f4e42edb8aa454ce241 https://doi.org/10.1584/jpestics.d20-23 Zobrazit plný text záznamu Plný text ve formátu PDF