The nucleotide sequences of the genes encoding the F1F0-ATPase beta-subunit from Oenococcus oeni, Leuconostoc mesenteroides subsp. mesenteroides, Pediococcus damnosus, Pediococcus parvulus, Lactobacillus brevis and Lactobacillus hilgardii were determined. Their deduced amino acid sequences showed homology values of 79-98%. Data from the alignment and ATPase tree indicated that O. oeni and L. mesenteroides subsp. mesenteroides formed a group well-separated from P. damnosus and P. parvulus and from the group comprises L. brevis and L. hilgardii. The N-terminus of the F1F0-ATPase beta-subunit of O. oeni contains a stretch of additional 38 amino acid residues. The catalytic site of the ATPase beta-subunit of the investigated strains is characterized by the two conserved motifs GGAGVGKT and GERTRE. The amplified atpD coding sequences were inserted into the pCRT7/CT-TOPO vector using TA-cloning strategy and transformed in Escherichia coli. SDS-PAGE and Western blot analyses confirmed that O. oeni has an ATPase beta-subunit protein which is larger in size than the corresponding molecules from the investigated strains.

中文翻译：

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葡萄酒乳酸菌中F1F0-ATPaseβ亚基的克隆，序列分析和表达。

猪肠球菌，Mesenteroides亚种的F1F0-ATPaseβ-亚基的基因的核苷酸序列。确定了肠系膜内膜炎，鼠李糖球菌，小球囊球菌，短乳杆菌和hilgardii乳杆菌。他们推导的氨基酸序列显示出79-98％的同源性值。来自比对和ATP酶树的数据表明O. oeni和L. mesenteroides亚种。mesenteroides形成了一个组，该组与damnosus damnosus和P. parvulus完全分离，并且由短短乳杆菌L. hilgardii组成。O. oeni的F1F0-ATPaseβ-亚基的N端含有一段额外的38个氨基酸残基。被研究菌株的ATPaseβ-亚基的催化位点由两个保守的基序GGAGVGKT和GERTRE表征。使用TA克隆策略将扩增的atpD编码序列插入到pCRT7 / CT-TOPO载体中，并在大肠杆菌中转化。SDS-PAGE和Western印迹分析证实o。oeni具有ATPaseβ亚基蛋白，其大小比来自研究菌株的相应分子大。