Cloning, sequence analysis and expression of the F1F0-ATPase beta-subunit from wine lactic acid bacteria.
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.
Sievers
Martin
M
Hochschule Wädenswil, Molekularbiologie, Wädenswil, Switzerland. m.sievers@hsw.ch
Uermösi
Christina
C
Fehlmann
Marc
M
Krieger
Sibylle
S
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000302:isolation & purification; D025243:Bacterial Proton-Translocating ATPases / Q000235:genetics; D015153:Blotting, Western; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005516:Food Microbiology; D006094:Gram-Positive Bacteria / Q000201:enzymology* / Q000235:genetics*; D019344:Lactic Acid / Q000096:biosynthesis*; D007778:Lactobacillus / Q000201:enzymology / Q000235:genetics; D007934:Leuconostoc / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D010376:Pediococcus / Q000201:enzymology / Q000235:genetics; D010802:Phylogeny; D016415:Sequence Alignment; D014920:Wine / Q000382:microbiology*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D019344:Lactic Acid; D025243:Bacterial Proton-Translocating ATPases