Tamaki T, Fukaya M, Takemura H, Tayama K, Okumura H, Kawamura Y, Nishiyama M, Horinouchi S, Beppu T
Nakano Central Research Institute, Nakano Vinegar Co., Ltd., Handa, Aichi, Japan.
Biochim Biophys Acta. 1991 Feb 16;1088(2):292-300. doi: 10.1016/0167-4781(91)90066-u.
The membrane-bound alcohol dehydrogenase (ADH) from Acetobacter polyoxogenes NBI1028 is composed of a 72 kDa subunit and a 44 kDa cytochrome c subunit. The amino acid sequences of the two regions of the 72 kDa subunit were determined to prepare oligonucleotides for the purpose of amplification of a DNA fragment corresponding to the intermediate region by the polymerase chain reaction. A 0.5 kb DNA fragment thus amplified was used as the probe to clone a 7.0 kb PstI fragment coding for the whole 72 kDa subunit. Nucleotide sequencing and immunoblot analysis revealed that the cloned fragment contained the full structural genes for the 72 kDa and the 44 kDa subunits and they were clustered with the same transcription polarity. The predicted amino acid sequence of the gene for the 72 kDa subunit showed homology with that of the 72 kDa subunit from ADH of A. aceti and those of methanol dehydrogenase from methylotrophic bacteria. The 72 and 44 kDa subunits contained one and three typical haem binding sequences, respectively.
多氧醋酸杆菌NBI1028的膜结合乙醇脱氢酶（ADH）由一个72 kDa亚基和一个44 kDa细胞色素c亚基组成。测定了72 kDa亚基两个区域的氨基酸序列，以制备寡核苷酸，用于通过聚合酶链反应扩增对应于中间区域的DNA片段。如此扩增得到的一个0.5 kb DNA片段用作探针，克隆编码整个72 kDa亚基的一个7.0 kb PstI片段。核苷酸测序和免疫印迹分析表明，克隆片段包含72 kDa和44 kDa亚基的完整结构基因，它们以相同的转录极性成簇。72 kDa亚基基因的预测氨基酸序列与醋酸醋杆菌ADH的72 kDa亚基以及甲基营养细菌甲醇脱氢酶的氨基酸序列具有同源性。72 kDa和44 kDa亚基分别包含一个和三个典型的血红素结合序列。
