Oxygen-dependent regulation of the expression of the catalase gene katA of Lactobacillus sakei LTH677.

Abstract

The catalase gene katA of Lactobacillus sakei LTH677 was cloned and expressed in Escherichia coli UM2, Lactobacillus casei LK1, and Lactobacillus curvatus LTH1432. The last host is a catalase-deficient plasmid-cured derivative of a starter organism used in meat fermentation. The regulation of katA expression was found to be the same in L. sakei LTH677 and the recombinant strains. The addition of H2O2 to anaerobic cultures, as well as a switch to aerobic conditions, resulted in a strong increase in KatA activity. The expression was investigated in more detail with L. sakei LTH677 and L. curvatus LTH4002. The recombinant strain LTH4002 did not accumulate H2O2 under glucose-limited aerobic conditions and remained viable in the stationary phase. Under inductive conditions, the katA-specific mRNA and the apoenzyme were synthesized de novo. Deletion derivatives of the katA promoter were produced, and the regulatory response was investigated by fusion to the beta-glucuronidase reporter gene gusA and expression in L. sakei LTH677. The fact that gene expression was subject to induction was confirmed at the level of transcription and protein synthesis. A small putative regulatory sequence of at least 25 bp was identified located upstream of the -35 site. Competition experiments performed with L. sakei LTH677 harboring the fusion constructs consisting of the katA promoter and gusA revealed that an activator protein is involved in the transcriptional induction of katA.