Duine J A, Frank J, Berkhout M P
FEBS Lett. 1984 Mar 26;168(2):217-21. doi: 10.1016/0014-5793(84)80249-5.
Cell-free extracts of methanol-grown Nocardia sp. 239 only show significant dye-linked methanol-oxidizing activity when NAD+ is added to the assay mixture. This activity resides in a multienzyme complex which could be resolved into 3 components, namely the methanol dehydrogenase, NAD-dependent aldehyde dehydrogenase and NADH dehydrogenase. In its dissociated form, the methanol dehydrogenase no longer shows dye reduction and although rises in the absorbance values around 340 nm are seen on addition of methanol plus NAD+ to the enzyme, this is not due to NADH production. However, dye reduction (NAD dependent) could be restored on incubating methanol dehydrogenase with the corresponding NADH dehydrogenase, obtained from the enzyme complex. It is concluded that this novel methanol dehydrogenase transfers the reducing equivalents, derived from methanol, directly to its associated NADH dehydrogenase via a mechanism in which NAD+ and PQQ are involved.
甲醇培养的诺卡氏菌属239的无细胞提取物,只有在向测定混合物中添加NAD⁺时,才表现出显著的与染料偶联的甲醇氧化活性。该活性存在于一种多酶复合物中,该复合物可分解为3个组分,即甲醇脱氢酶、NAD依赖的醛脱氢酶和NADH脱氢酶。以解离形式存在时,甲醇脱氢酶不再表现出染料还原作用,并且尽管在向该酶中添加甲醇和NAD⁺后,在340nm左右的吸光度值会升高,但这并非由于产生了NADH。然而,将甲醇脱氢酶与从酶复合物中获得的相应NADH脱氢酶一起孵育时,染料还原作用(NAD依赖)可以恢复。得出的结论是,这种新型甲醇脱氢酶通过一种涉及NAD⁺和PQQ的机制,将源自甲醇的还原当量直接转移至其相关的NADH脱氢酶。