Lysis of an Lactococcus lactis Di-peptidase Mutant and Rescue by Mutation in the Pleiotropic Regulator CodY

Huang, C., Hernandez-Valdes, J. A., Kuipers, O. P. & Kok, J., 31-Jan-2020, In : Applied and environmental microbiology.

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Lactococcus lactis subsp. cremoris MG1363 is a model for the lactic acid bacteria (LAB) used in the dairy industry. The proteolytic system, consisting of a proteinase, several peptide- and amino acid uptake systems and a host of intracellular peptidases, plays a vital role in nitrogen metabolism and is of eminent importance for flavor formation in dairy products. The dipeptidase PepV functions in the last stages of proteolysis. A link between nitrogen metabolism and peptidoglycan (PG) biosynthesis was underlined by the finding that deletion of the dipeptidase gene pepV (MGΔpepV) results in a prolonged lag phase when the mutant strain was grown with a high concentration of glycine. In addition, most MGΔpepV cells lyse and have serious defects in their shape. This phenotype is due to a shortage of alanine, since adding alanine can rescue the growth and shape defects. Strain MGΔpepV is more resistant to vancomycin, an antibiotic targeting peptidoglycan D-Ala-D-Ala ends, which confirmed that MGΔpepV has an abnormal PG composition. A mutant of MGΔpepV was obtained in which growth inhibition and cell shape defects were alleviated. Genome sequencing shows that this mutant has a single point mutation in the codY gene, resulting in an arginine residue at position 218 in the DNA-binding motif of CodY being replaced by a cysteine residue. Thus, this strain was named MGΔpepV_codYR218C RNA-seq data reveals a dramatic de-repression in peptide uptake and amino acid utilization in MGΔpepV_codYR218C Thus, a model of the connections among PepV activity, CodY regulation, and PG synthesis of L. lactis is proposed.ImportancePrecise control of peptidoglycan synthesis is essential in Gram-positive bacteria for maintaining cell shape and integrity as well as resisting stresses. Although neither the dipeptidase PepV nor alanine is essential for L. lactis MG1363, adequate availability of either ensures proper cell wall synthesis. We broaden the knowledge about the dipeptidase PepV, which acts as a linker between nitrogen metabolism and cell wall synthesis in L. lactis.

Original languageEnglish
JournalApplied and environmental microbiology
Publication statusE-pub ahead of print - 31-Jan-2020

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