Jensen Jr Kenneth A, Ryan Zachary C, Vanden Wymelenberg Amber, Cullen Daniel, Hammel Kenneth E
Institute for Microbial and Biochemical Technology, USDA Forest Products Laboratory, Madison, Wisconsin 53705, USA.
Appl Environ Microbiol. 2002 Jun;68(6):2699-703. doi: 10.1128/AEM.68.6.2699-2703.2002.
The brown-rot basidiomycete Gloeophyllum trabeum uses a quinone redox cycle to generate extracellular Fenton reagent, a key component of the biodegradative system expressed by this highly destructive wood decay fungus. The hitherto uncharacterized quinone reductase that drives this cycle is a potential target for inhibitors of wood decay. We have identified the major quinone reductase expressed by G. trabeum under conditions that elicit high levels of quinone redox cycling. The enzyme comprises two identical 22-kDa subunits, each with one molecule of flavin mononucleotide. It is specific for NADH as the reductant and uses the quinones produced by G. trabeum (2,5-dimethoxy-1,4-benzoquinone and 4,5-dimethoxy-1,2-benzoquinone) as electron acceptors. The affinity of the reductase for these quinones is so high that precise kinetic parameters were not obtainable, but it is clear that k(cat)/K(m) for the quinones is greater than 10(8) M(-1) s(-1). The reductase is encoded by a gene with substantial similarity to NAD(P)H:quinone reductase genes from other fungi. The G. trabeum quinone reductase may function in quinone detoxification, a role often proposed for these enzymes, but we hypothesize that the fungus has recruited it to drive extracellular oxyradical production.
褐腐担子菌变色栓菌利用醌氧化还原循环来产生细胞外芬顿试剂,这是这种极具破坏性的木材腐朽真菌所表达的生物降解系统的关键组成部分。驱动该循环的迄今未被表征的醌还原酶是木材腐朽抑制剂的潜在靶点。我们已经鉴定出在引发高水平醌氧化还原循环的条件下变色栓菌所表达的主要醌还原酶。该酶由两个相同的22 kDa亚基组成,每个亚基含有一分子黄素单核苷酸。它对作为还原剂的NADH具有特异性,并利用变色栓菌产生的醌(2,5 - 二甲氧基 - 1,4 - 苯醌和4,5 - 二甲氧基 - 1,2 - 苯醌)作为电子受体。该还原酶对这些醌的亲和力非常高,以至于无法获得精确的动力学参数,但很明显,醌的k(cat)/K(m)大于10(8) M(-1) s(-1)。该还原酶由一个与其他真菌的NAD(P)H:醌还原酶基因具有高度相似性的基因编码。变色栓菌醌还原酶可能在醌解毒中发挥作用,这是这些酶常被提出的作用,但我们推测该真菌已将其招募来驱动细胞外氧自由基的产生。