Yamamoto I, Ishimoto M
J Biochem. 1978 Sep;84(3):673-9. doi: 10.1093/oxfordjournals.jbchem.a132172.
E. coli K10 was found to grow anaerobically on molecular hydrogen by reducing nitrate, fumarate, and trimethylamine N-oxide when peptone was added to the culture medium. Molar growth yields based on consumed hydrogen estimated from the amounts of reduction products were all 7.8 g cells/mol, suggesting that 1 mol of ATP was produced in the oxidation of 1 mol of hydrogen. Hydrogenase activity measured in terms of hydrogen evolution was several times higher in cells grown on glucose than in cells grown on hydrogen in the presence of fumarate and trimethylamine N-oxide, while hydrogenase activity measured in terms of hydrogen uptake was unchanged in both cases. The ratio of hydrogenase activities measured in terms of hydrogen uptake and evolution was also high in the extract and centrifugal fractions from cells grown in hydrogen. The soluble fraction and trypsin digest of the precipitate at 100,000 X g were subjected to polyacrylamide disc gel electrophoresis and hydrogenase bands were stained by reduction of benzyl viologen with hydrogen and by oxidation of reduced methyl viologen. The resulting patterns suggest that multiple forms of hydrogenase are present and that the amounts of forms functioning in hydrogen evolution were greatly decresed in cells grown on hydrogen in the presence of acceptors.
当向培养基中添加蛋白胨时，发现大肠杆菌K10能够通过还原硝酸盐、富马酸盐和三甲胺N-氧化物在分子氢上厌氧生长。根据还原产物的量估算，基于消耗的氢的摩尔生长产率均为7.8克细胞/摩尔，这表明在1摩尔氢的氧化过程中产生了1摩尔ATP。以氢气释放量衡量的氢化酶活性，在以葡萄糖为碳源生长的细胞中比在存在富马酸盐和三甲胺N-氧化物时以氢气为碳源生长的细胞中高几倍，而以氢气吸收量衡量的氢化酶活性在两种情况下均未改变。在以氢气为碳源生长的细胞的提取物和离心级分中，以氢气吸收和释放量衡量的氢化酶活性之比也很高。将100,000×g下沉淀的可溶性级分和胰蛋白酶消化物进行聚丙烯酰胺圆盘凝胶电泳，并用氢气还原苄基紫精和氧化还原型甲基紫精对氢化酶条带进行染色。所得图谱表明存在多种形式的氢化酶，并且在存在受体的情况下以氢气为碳源生长的细胞中，参与氢气释放的氢化酶形式的量大大减少。
