Regulation and adaptive evolution of lactose operon expression in Lactobacillus delbrueckii.
Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis are both used in the dairy industry as homofermentative lactic acid bacteria in the production of fermented milk products. After selective pressure for the fast fermentation of milk in the manufacture of yogurts, L. delbrueckii subsp. bulgaricus loses its ability to regulate lac operon expression. A series of mutations led to the constitutive expression of the lac genes. A complex of insertion sequence (IS) elements (ISL4 inside ISL5), inserted at the border of the lac promoter, induced the loss of the palindromic structure of one of the operators likely involved in the binding of regulatory factors. A lac repressor gene was discovered downstream of the beta-galactosidase gene of L. delbrueckii subsp. lactis and was shown to be inactivated by several mutations in L. delbrueckii subsp. bulgaricus. Regulatory mechanisms of the lac gene expression of L. delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis were compared by heterologous expression in Lactococcus lactis of the two lac promoters in front of a reporter gene (beta-glucuronidase) in the presence or absence of the lac repressor gene. Insertion of the complex of IS elements in the lac promoter of L. delbrueckii subsp. bulgaricus increased the promoter's activity but did not prevent repressor binding; rather, it increased the affinity of the repressor for the promoter. Inactivation of the lac repressor by mutations was then necessary to induce the constitutive expression of the lac genes in L. delbrueckii subsp. bulgaricus.
Lapierre
Luciane
L
Nestlé Research Center, Nestlé, Ltd., CH-1000 Lausanne 26, Switzerland.
Mollet
Beat
B
Germond
Jacques-Edouard
JE
D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D001483:Base Sequence; D004269:DNA, Bacterial; D019143:Evolution, Molecular; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D007763:Lac Operon; D007778:Lactobacillus / Q000235:genetics*; D007785:Lactose / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D001616:beta-Galactosidase / Q000235:genetics
D004269:DNA, Bacterial; D001616:beta-Galactosidase; D007785:Lactose