Linke Diana, Lehnert Nicole, Nimtz Manfred, Berger Ralf G
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, D-30167, Hannover, Germany.
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, D-30167, Hannover, Germany.
Enzyme Microb Technol. 2014 Jul-Aug;61-62:7-12. doi: 10.1016/j.enzmictec.2014.04.001. Epub 2014 Apr 23.
An intracellular alcohol oxidase (AOX) was isolated from the white-rot basidiomycete Phanerochaete chrysosporium (Pch), grown on l-lactate induction medium, and purified to electrophoretic homogeneity. The dimeric protein consisted of two identical 75kDa subunits. The open reading frame of 1,956bp resulted in a monomer consisting of 651 amino acids. The enzyme showed a pI at 5.4, a pH optimum of 9, a temperature optimum at 50°C, possessed putative conserved domains of the GMC superfamily, a FAD binding domain, and showed up to 86% homology to alcohol oxidase sequences of Gloeophyllum trabeum and Coprinopsis cinerea. As was shown for the first time for an AOX from a basidiomycete, not only methanol, but also lower primary alcohols and glycerol were accepted as substrates. An assay based on aldehyde dehydrogenase confirmed d-glyceraldehyde as the product of the reaction. A bioprocess based on this enzyme could alleviate the problems associated with the huge side-stream of glycerol occurring during the manufacture of biodiesel, yielding the green oxidant hydrogen peroxide.
从在L-乳酸诱导培养基上生长的白腐担子菌黄孢原毛平革菌（Pch）中分离出一种细胞内乙醇氧化酶（AOX），并将其纯化至电泳纯。该二聚体蛋白由两个相同的75kDa亚基组成。1956bp的开放阅读框产生了一个由651个氨基酸组成的单体。该酶的pI为5.4，最适pH为9，最适温度为50°C，具有GMC超家族的推定保守结构域、一个FAD结合结构域，与密粘褶菌和灰盖鬼伞的乙醇氧化酶序列具有高达86%的同源性。正如首次在担子菌的AOX中所显示的那样，不仅甲醇，而且低级伯醇和甘油也被接受为底物。基于醛脱氢酶的测定证实d-甘油醛是该反应的产物。基于这种酶的生物过程可以缓解与生物柴油生产过程中产生的大量甘油副产物相关的问题，产生绿色氧化剂过氧化氢。
