Identification of a gene cluster encoding Krebs cycle oxidative enzymes linked to the pyruvate carboxylase gene in Lactococcus lactis ssp. lactis C2.
We identified a 14-kb pyruvate carboxylase gene-containing fragment from a lactococcal C2-lambda phage genomic library. Downstream of the pyruvate carboxylase gene-containing fragment, a gene cluster coding for open reading frames displaying extensive homology to citrate synthase, aconitase, and a truncated isocitrate dehydrogenase was identified. However, the truncation was shown to have occurred during the cloning by two noncontiguous Sau3AI fragments ligating together. The lactococcal citrate synthase gene consisted of 1323 bp and encoded a 441-amino acid citrate synthase protein. The lactococcal aconitase gene was 2544 bp and encoded an 848-amino acid protein. Corresponding to the complete citrate synthase gene, citrate synthase activity was detected in Lactococcus lactis ssp. lactis C2. Isocitrate dehydrogenase activity was found to be missing in Lactococcus lactis C2, suggesting that the gene may be incomplete or is not expressed, resulting in a requirement for glutamic acid in lactococci.
Wang
H
H
Department of Food Science and Nutrition University of Minnesota, St. Paul 55108, USA.
Baldwin
K A
KA
O'Sullivan
D J
DJ
McKay
L L
LL
D000154:Aconitate Hydratase / Q000235:genetics; D002950:Citrate (si)-Synthase / Q000235:genetics; D002952:Citric Acid Cycle; D005798:Genes, Bacterial; D008040:Genetic Linkage; D007521:Isocitrate Dehydrogenase / Q000235:genetics; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D011766:Pyruvate Carboxylase / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D007521:Isocitrate Dehydrogenase; D002950:Citrate (si)-Synthase; D000154:Aconitate Hydratase; D011766:Pyruvate Carboxylase