De Bruyn J, Johannes A, Weckx M, Beumer-Jochmans M P
J Gen Microbiol. 1981 Jun;124(2):359-63. doi: 10.1099/00221287-124-2-359.
An alcohol dehydrogenase of broad specificity was purified 43-fold from extracts of Mycobacterium tuberculosis var. bovis (BCG) grown on Sauton medium. It was also present in cells grown in Dubos medium and Tween 80 and bovine serum albumin. The enzyme, which appeared to be soluble, acted as an oxidoreductase in the system butan-1-ol-NADP. It was eluted from Sephadex G-200, hydroxylapatite and DEAE-cellulose in a single peak. The molecular weight, as determined by gel filtration on Sephadex G-200, was about 75,000. Results of electrophoresis in sodium dodecyl sulphate-polyacrylamide gels were compatible with the existence of two subunits each of molecular weight 37,500. The optimum pH was about 8.5 when the enzyme catalysed the oxidation of butan-1-ol, and about 8.2 for the reverse reaction. The apparent Km was 0.125 mM for butyraldehyde and 0.22 M for butan-1-ol. The dehydrogenase activity was maintained after heat treatment (40 min at 55 degrees C) in the presence of 30% (W/V) glycerol, but was abolished by heating (40 min at 55 degrees C) in the presence of 0.1 M-EDTA. The activity of enzyme inactivated by heat and EDTA could be fully restored at room temperature in the presence of 2 mM-Zn2+.
从在索顿培养基上生长的牛型结核分枝杆菌（卡介苗）提取物中纯化出一种具有广泛特异性的乙醇脱氢酶，纯化倍数为43倍。在杜博斯培养基、吐温80和牛血清白蛋白中生长的细胞中也存在该酶。该酶似乎是可溶的，在丁醇 - NADP系统中作为氧化还原酶起作用。它在葡聚糖凝胶G - 200、羟基磷灰石和DEAE - 纤维素上以单一峰被洗脱。通过葡聚糖凝胶G - 200凝胶过滤测定的分子量约为75,000。在十二烷基硫酸钠 - 聚丙烯酰胺凝胶中的电泳结果与存在两个分子量均为37,500的亚基相一致。当该酶催化丁醇氧化时，最适pH约为8.5，逆反应的最适pH约为8.2。丁醛的表观Km为0.125 mM，丁醇的表观Km为0.22 M。在30%（W/V）甘油存在下热处理（55℃ 40分钟）后，脱氢酶活性得以维持，但在0.1 M - EDTA存在下加热（55℃ 40分钟）则会使其失活。在2 mM - Zn2+存在下，室温下热和EDTA失活的酶的活性可完全恢复。
