Cloning, sequence analysis and expression of the F1F0-ATPase beta-subunit from wine lactic acid bacteria |
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Authors: | Sievers Martin Uermösi Christina Fehlmann Marc Krieger Sibylle |
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Affiliation: | Hochschule W?denswil, Molekularbiologie, W?denswil, Switzerland. m.sievers@hsw.ch |
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Abstract: | 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. |
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Keywords: | F1F0-ATPase β -subunit atpD lactic acid bacteria TA-cloning |
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