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

Wu, Hao; Zhao, Yue; Du, Yuhui; Miao, Sen; Liu, Jingui; Li, Yanni; Caiyin, Qinggele; Qiao, Jianjun

doi:10.3168/jds.2018-14594

Cited by 24 publications

(23 citation statements)

References 69 publications

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“…We speculated that the improved multi-stress tolerance of the mutant strains was owing to the loss of PRF, resulting in remodeling of the cell wall/ membrane and changes in carbon source metabolism. Previous studies have shown that the structure of the cell wall/membrane affects the nisin tolerance of LAB [45], and cell wall, as the first barrier of bacteria, is essential for the strain's stress tolerance [20]. Similarly, in the present study, the synthesis levels of enzymes related to cell wall/membrane biosynthesis and degradation in L. lactis N8-1 were noted to be altered.…”

Section: Discussionsupporting

confidence: 75%

“…In general, deletion of non-essential genome fragments, such as prophages and transposons, could confer some advantages on bacterial strains [17], including enhanced growth, increased biomass, and higher level of proteins synthesis [18]. Besides, prophages can usually synthesize proteins like prophage lysin that affect the host cell wall/membrane [19], and cell wall/membrane is essential for maintaining cellular integrity and resisting environmental stress [20]. For instance, the structure of the cell wall/membrane of nisin-producing L. lactis can also affect its tolerance to nisin [21].…”

Section: Introductionmentioning

confidence: 99%

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Engineering Lactococcus lactis as a multi-stress tolerant biosynthetic chassis by deleting the prophage-related fragment

Qiao

Liu

et al. 2020

Microb Cell Fact

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Background In bioengineering, growth of microorganisms is limited because of environmental and industrial stresses during fermentation. This study aimed to construct a nisin-producing chassis Lactococcus lactis strain with genome-streamlined, low metabolic burden, and multi-stress tolerance characteristics. Results The Cre-loxP recombination system was applied to reduce the genome and obtain the target chassis strain. A prophage-related fragment (PRF; 19,739 bp) in the L. lactis N8 genome was deleted, and the mutant strain L. lactis N8-1 was chosen for multi-stress tolerance studies. Nisin immunity of L. lactis N8-1 was increased to 6500 IU/mL, which was 44.44% higher than that of the wild-type L. lactis N8 (4500 IU/mL). The survival rates of L. lactis N8-1 treated with lysozyme for 2 h and lactic acid for 1 h were 1000- and 10,000-fold higher than that of the wild-type strain, respectively. At 39 ℃, the L. lactis N8-1 could still maintain its growth, whereas the growth of the wild-type strain dramatically dropped. Scanning electron microscopy showed that the cell wall integrity of L. lactis N8-1 was well maintained after lysozyme treatment. Tandem mass tags labeled quantitative proteomics revealed that 33 and 9 proteins were significantly upregulated and downregulated, respectively, in L. lactis N8-1. These differential proteins were involved in carbohydrate and energy transport/metabolism, biosynthesis of cell wall and cell surface proteins. Conclusions PRF deletion was proven to be an efficient strategy to achieve multi-stress tolerance and nisin immunity in L. lactis, thereby providing a new perspective for industrially obtaining engineered strains with multi-stress tolerance and expanding the application of lactic acid bacteria in biotechnology and synthetic biology. Besides, the importance of PRF, which can confer vital phenotypes to bacteria, was established.

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Section: Discussionsupporting

confidence: 75%

Section: Introductionmentioning

confidence: 99%

Engineering Lactococcus lactis as a multi-stress tolerant biosynthetic chassis by deleting the prophage-related fragment

Qiao

Liu

et al. 2020

Microb Cell Fact

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“…Under carbohydrate-limiting conditions, homofermentative LAB metabolize pyruvate via pyruvate formate-lyase or pyruvate dehydrogenase to formate/CO 2 , acetate, and ethanol as an alternative to lactate formation (24), with pyruvate formate-lyase being preferentially used under anaerobic conditions at nearneutral pH (59,60). In addition, downregulation of L-lactate dehydrogenase and upregulation of pyruvate formate-lyase in response to low pH and high lactate concentrations were observed in Lactococcus lactis (61). The lack of lactate production by Carnobacterium spp.…”

Section: Discussionmentioning

confidence: 99%

Complementary Antibacterial Effects of Bacteriocins and Organic Acids as Revealed by Comparative Analysis of Carnobacterium spp. from Meat

Zhang

Gänzle

Yang

2019

Appl Environ Microbiol

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Carnobacterium maltaromaticum and Carnobacterium divergens are often predominant in the microbiota of vacuum-packaged (VP) meats after prolonged storage at chiller temperatures, and more so in recent studies. We investigated the antibacterial activities of C. maltaromaticum and C. divergens (n = 31) from VP meats by phenotypic characterization and genomic analysis. Five strains showed antibacterial activities against Gram-positive bacteria in a spot-lawn assay, with C. maltaromaticum strains having an intergeneric and C. divergens strains an intrageneric inhibition spectrum. This inhibitory activity is correlated with the production of predicted bacteriocins, including carnobacteriocin B2 and carnolysin for C. maltaromaticum and divergicin A for C. divergens. The supernatants of both species cultured in meat juice medium under anaerobic conditions retarded the growth of most Gram-positive and Gram-negative bacteria in broth assay in a strain-dependent manner. C. maltaromaticum and C. divergens produced formate and acetate but not lactate under VP meat-relevant conditions. The relative inhibitory activity by Carnobacterium strains was significantly correlated (P < 0.05) to the production of both acids. Genomic analysis revealed the presence of genes required for respiration in both species. In addition, two clusters of C. divergens have an average nucleotide identity below the cutoff value for species delineation and thus should be considered to be two subspecies. In conclusion, both bacteriocins and organic acids are factors contributing significantly to the antibacterial activity of C. maltaromaticum and C. divergens under VP meat-relevant conditions. A few Carnobacterium strains can be explored as protective cultures to extend the shelf life and improve the safety of VP meats. IMPORTANCE The results of this study demonstrated that both bacteriocins and organic acids are important factors contributing to the antibacterial activities of Carnobacterium from vacuum-packaged (VP) meats. This study demonstrated that formate and acetate are the key organic acids produced by Carnobacterium and demonstrated their association with the inhibitory activity of carnobacteria under VP meat-relevant storage conditions. The role of lactate, on the other hand, may not be as important as previously believed in the antimicrobial activities of Carnobacterium spp. on chilled VP meats. These findings advance our understanding of the physiology of Carnobacterium spp. to better explore their biopreservative properties for chilled VP meats.

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“…Figure 5A shows that arginine accounts for the highest proportion of the intracellular free amino acids. In fact, the arginine deaminase (ADI) system is considered to be an important factor to protect microbial cells against acidic environment (Senouci-Rezkallah et al, 2011;Wu et al, 2018). Besides, aspartate can be converted into arginine to participate in the ADI system, accompanied by the formation of NH 3 (Fernández and Zúñiga, 2006).…”

Section: Effects Of Environmental Ph On Intracellular Free Amino Acidmentioning

confidence: 99%

Physiological and Transcriptional Responses of Streptomyces albulus to Acid Stress in the Biosynthesis of ε-Poly-L-lysine

et al. 2020

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Streptomyces albulus has commercially been used for the production of ε-poly-Llysine (ε-PL), a natural food preservative, where acid stress is inevitably encountered in the biosynthesis process. To elucidate the acid tolerance response (ATR), a comparative physiology and transcriptomic analysis of S. albulus M-Z18 at different environmental pH (5.0, 4.0, and 3.0) was carried out. In response to acid stress, cell envelope regulated the membrane fatty acid composition and chain length to reduce damage. Moreover, intracellular pH homeostasis was maintained by increasing H +-ATPase activity and intracellular ATP and amino acid (mainly arginine, glutamate, aspartate and lysine) concentrations. Transcriptional analysis based on RNA-sequencing indicated that acid stress aroused global changes and the differentially expressed genes involved in transcriptional regulation, stress-response protein, transporter, cell envelope, secondary metabolite biosynthesis, DNA and RNA metabolism and ribosome subunit. Consequently, the ATR of S. albulus was preliminarily proposed. Notably, it is indicated that the biosynthesis of ε-PL is also a response mechanism for S. albulus to combat acid stress. These results provide new insights into the ATR of S. albulus and will contribute to the production of ε-PL via adaptive evolution or metabolic engineering.

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

Cited by 24 publications

References 69 publications

Engineering Lactococcus lactis as a multi-stress tolerant biosynthetic chassis by deleting the prophage-related fragment

Engineering Lactococcus lactis as a multi-stress tolerant biosynthetic chassis by deleting the prophage-related fragment

Complementary Antibacterial Effects of Bacteriocins and Organic Acids as Revealed by Comparative Analysis of Carnobacterium spp. from Meat

Physiological and Transcriptional Responses of Streptomyces albulus to Acid Stress in the Biosynthesis of ε-Poly-L-lysine

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