Abstract
BackgroundLactococcus lactis is industrially employed to manufacture various fermented dairy products. The most cost-effective method for the preservation of L. lactis starter cultures is spray drying, but during this process cultures encounter heat and oxidative stress, typically resulting in low survival rates. However, viability of starter cultures is essential for their adequate contribution to milk fermentation, supporting the ambition to better understand and improve their robustness phenotypes.ResultsThis study describes a transcriptome-phenotype matching approach in which the starter L. lactis MG1363 was fermented under a variety of conditions that differed in the levels of oxygen and/or salt, as well as the fermentation pH and temperature. Samples derived from these fermentations in the exponential phase of bacterial growth were analyzed by full-genome transcriptomics and the assessment of heat and oxidative stress phenotypes. Variations in the fermentation conditions resulted in up to 1000-fold differences in survival during heat and oxidative stress. More specifically, aeration during fermentation induced protection against heat stress, whereas a relatively high fermentation temperature resulted in enhanced robustness towards oxidative stress. Concomitantly, oxygen levels and fermentation temperature induced differential expression of markedly more genes when compared with the other fermentation parameters. Correlation analysis of robustness phenotypes and gene expression levels revealed transcriptome signatures for oxidative and/or heat stress survival, including the metC-cysK operon involved in methionine and cysteine metabolism. To validate this transcriptome-phenotype association we grew L. lactis MG1363 in the absence of cysteine which led to enhanced robustness towards oxidative stress.ConclusionsOverall, we demonstrated the importance of careful selection of fermentation parameters prior to industrial processing of starter cultures. Furthermore, established stress genes as well as novel genes were associated with robustness towards heat and/or oxidative stress. Assessment of the expression levels of this group of genes could function as an indicator for enhanced selection of fermentation parameters resulting in improved robustness during spray drying. The increased robustness after growth without cysteine appeared to confirm the role of expression of the metC-cysK operon as an indicator of robustness and suggests that sulfur amino acid metabolism plays a pivotal role in oxidative stress survival.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-014-0148-6) contains supplementary material, which is available to authorized users.
Highlights
Lactococcus lactis is industrially employed to manufacture various fermented dairy products
In this study, we demonstrated the importance of careful selection of fermentation parameters prior to industrial processing of starter cultures
We have shown that correlation of heat and oxidative stress survival phenotypes and gene expression levels can reveal transcriptome signatures for robustness
Summary
Lactococcus lactis is industrially employed to manufacture various fermented dairy products. Lactococcus lactis is important during milk fermentations, as this lactic acid bacterium (LAB) positively contributes to the preservation, flavor and texture characteristics of the resulting fermented end-products, which include cheese and butter(milk) [1]. Nowadays, these milk fermentation processes have largely been industrialized and standardized, and are initiated with the addition of starter cultures containing high concentrations of one or multiple L. lactis strains. Transcriptome analyses of strain MG1363 revealed the regulatory networks of nitrogen metabolism [CodY [10]], carbon catabolite repression [CcpA [11]] and fatty acid biosynthesis [FabT [12]], and this technology was employed to study gene expression during growth in milk [13]
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