Quantitative proteomics of Lactococcus lactis F44 under cross-stress of low pH and lactate

Abstract

Lactococcus lactis encounters 3 environmental stimuli, H⁺, lactate, and undissociated lactic acid, because of the accumulation of lactic acid-the predominant fermentation product. Few studies have examined how these stimuli synergistically affect the bacteria. Herein, we analyzed the dissociation degree of lactic acid at different pH and investigated the cellular response to cross-stress in L. lactis ssp. lactis F44 through quantitative proteomic analysis using isobaric tags for relative and absolute quantitation of 3 groups: 0% lactic acid with pH 4.0 and 0% lactic acid with pH 5.0 for acid stress; 2% lactic acid with pH 7.0 and 3% lactic acid with pH 7.0 for lactate stress; and 2% lactic acid with pH 4.0, 2% lactic acid with pH 5.0, 3% lactic acid with pH 4.0, and 3% lactic acid with pH 5.0 for cross-stress. We observed that the metabolisms of carbohydrate and energy were inhibited, which might be due to the feedback inhibition of lactic acid. The arginine deiminase pathway was improved to maintain the stability of intracellular pH. Additionally, some differentially expressed genes associated with the general stress response, amino acid metabolism, cell wall synthesis, and regulatory systems played significant roles in stress response. Overall, we highlighted the response mechanisms to lactic acid stress and provided a new opportunity for constructing robust industrial strains.

Keywords: Lactococcus lactis; quantitative proteomics; stress response.