Enhancing survival of Escherichia coli by expression of azoreductase AZR possessing quinone reductase activity.
Quinone reductase activity of azoreductase AZR from Rhodobacter sphaeroides was reported. High homologies were found in the cofactor/substrate-binding regions of quinone reductases from different domains. 3D structure comparison revealed that AZR shared a common overall topology with mammal NAD(P)H/quinone oxidoreductase NQO1. With menadione as substrate, the optimal pH value and temperature were pH 8-9 and 50 degrees C, respectively. Following the ping-pong kinetics, AZR transferred two electrons from NADPH to quinone substrate. It could reduce naphthoquinones and anthraquinones, such as menadione, lawsone, anthraquinone-2-sulfonate, and anthraquinone-2,6-disulfonate. However, no activity was detected with 1,4-benzoquinone. Dicoumarol competitively inhibited AZR's quinone reductase activity with respect to NADPH, with an obtained K (i) value of 87.6 microM. Significantly higher survival rates were obtained in Escherichia coli YB overexpressing AZR than in the control strain when treated by heat shock and oxidative stressors such as H(2)O(2) and menadione.
Liu
Guangfei
G
School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, People's Republic of China.
Zhou
Jiti
J
Jin
Ruofei
R
Zhou
Mi
M
Wang
Jing
J
Lu
Hong
H
Qu
Yuanyuan
Y
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D050296:Microbial Viability; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009601:Nitroreductases; D012242:Rhodobacter sphaeroides / Q000201:enzymology*; D016415:Sequence Alignment; D013379:Substrate Specificity
D001426:Bacterial Proteins; D009247:NADH, NADPH Oxidoreductases; D016660:NAD(P)H Dehydrogenase (Quinone); D009601:Nitroreductases; C021106:azoreductase