title: Effect of redox mediators on nitrogenase and hydrogenase activities in Azotobacter vinelandii.
abstract: In bioelectrochemical studies, redox mediators such as methylene blue, natural red, and thionine are used to studying the redox characteristics of enzymes in the living cell. Here we show that nitrogenase activity in Azotobacter vinelandii is completely inhibited by oxidized methylene blue (MBo) when the concentration of this mediator in the medium is increased up to 72 microM. This activity in A. vinelandii is somewhat inhibited by a coenzyme, ascorbic acid (AA). However, the nitrogenase activity within the A. vinelandii cell is unchanged even for a high concentration of oxidized natural red (NRo) alone. Interestingly, these mediators and AA do not have the capacity to inhibit the H2 uptake activity of the hydrogenase in A. vinelandii. Average active rates of 66 nM H2 evolved/mg cell protein/min from the nitrogenase and 160 nM H2-uptake/mg cell protein/min from the hydrogenase in A. vinelandii are found in aid of the activities of the enzymes for H2 evolution and for H2 uptake are compared. The activities of both enzymes in A. vinelandii are strongly inhibited by thionine having high oxidative potential. Mechanisms of various mediators acting in vivo for both enzymes in A. vinelandii are discussed.
authors: School of Life Sciences, Xiamen University, China. hqhuang@jingxian.xmu.edu.cn
authors: H Q
authors: HQ
authors: Huang
authors: Q M
authors: QM
authors: Lin
authors: W J
authors: WJ
authors: Zhai
authors: C H
authors: CH
authors: Chen
mesh_terms: D016948:Azotobacter vinelandii / Q000187:drug effects* / Q000201:enzymology; D004396:Coloring Agents / Q000494:pharmacology*; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009591:Nitrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010084:Oxidation-Reduction
chemical_list: D004396:Coloring Agents; D006859:Hydrogen; D006864:Hydrogenase; D009591:Nitrogenase