STUDIES OF CONTROL OF LUMINESCENCE IN BENECKEA-HARVEYI - PROPERTIES OF NADH AND NADPH-FMN OXIDOREDUCTASES
STUDIES OF CONTROL OF LUMINESCENCE IN BENECKEA-HARVEYI - PROPERTIES OF NADH AND NADPH-FMN OXIDOREDUCTASES
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DOI：
10.1021/bi00597a018
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发表时间：
1978-01-01
期刊：
影响因子：
2.9
通讯作者：
DELUCA, M
中科院分区：
文献类型：
--
作者：
JABLONSKI, E;DELUCA, M
来自B的高度纯化的NADH和NADPH：FMN氧化还原酶。harveyi的动力学参数、与荧光素酶的结合、与人工电子受体的活性和抑制剂的作用。NADH：FMN氧化还原酶表现出单置换动力学，而NADPH：FMN氧化还原酶表现出双置换或乒乓动力学。这与NADPH：FMN氧化还原酶催化的反应中作为中间体的还原酶的形成一致。任何一种氧化还原酶与荧光素酶反应的偶联降低了NADH、NADPH和FMN的表观Km，支持氧化还原酶和荧光素酶之间复合物的建议。可溶性氧化还原酶在用荧光素酶产生光方面比从这些细菌的膜获得的NADH脱氢酶制剂更有效。可溶性酶使用FMN或FAD作为还原吡啶核苷酸氧化的底物，而膜NADH脱氢酶对人工电子受体如铁氰化物和亚甲蓝更有活性。FMN和FAD是非常差的受体。显然，两种可溶性氧化还原酶都不是来自膜。这两种酶都是组成型的，不依赖于荧光素酶的合成。
Highly purified NADH and NADPH:FMN oxidoreductases from B. harveyi were characterized with regard to kinetic parameters, association with luciferase, activity with artificial electron acceptors and the effects of inhibitors. The NADH:FMN oxidoreductase exhibits single displacement kinetics while the NADPH:FMN oxidoreductase exhibits double displacement or ping-pong kinetics. This is consistent with the formation of a reduced enzyme as an intermediate in the reaction catalyzed by the NADPH:FMN oxidoreductase. Coupling of either of the oxidoreductases to the luciferase reaction decreases the apparent Km for NADH, NADPH and FMN, supporting the suggestion of a complex between the oxidoreductases and luciferase. The soluble oxidoreductases are more efficient in producing light with luciferase than is a NADH dehydrogenase preparation obtained from the membranes of these bacteria. The soluble enzymes use FMN or FAD as substrates for the oxidation of reduced pyridine nucleotides while the membrane NADH dehydrogenase is much more active with artificial electron acceptors such as ferricyanide and methylene blue. FMN and FAD are very poor acceptors. Apparently, neither of the soluble oxidoreductases is derived from the membranes. Both enzymes are constitutive and do not depend on the synthesis of luciferase.