Quantitative Proteomics for the Comprehensive Analysis of Stress Responses of Lactobacillus paracasei subsp. paracasei F19

Abstract

Lactic acid bacteria are broadly employed as starter cultures in the manufacture of foods. Upon technological preparation, they are confronted with drying stress that amalgamates numerous stress conditions resulting in losses of fitness and survival. To better understand and differentiate physiological stress responses, discover general and specific markers for the investigated stress conditions, and predict optimal preconditioning for starter cultures, we performed a comprehensive genomic and quantitative proteomic analysis of a commonly used model system, Lactobacillus paracasei subsp. paracasei TMW 1.1434 (isogenic with F19) under 11 typical stress conditions, including among others oxidative, osmotic, pH, and pressure stress. We identified and quantified >1900 proteins in triplicate analyses, representing 65% of all genes encoded in the genome. The identified genes were thoroughly annotated in terms of subcellular localization prediction and biological functions, suggesting unbiased and comprehensive proteome coverage. In total, 427 proteins were significantly differentially expressed in at least one condition. Most notably, our analysis suggests that optimal preconditioning toward drying was predicted to be alkaline and high-pressure stress preconditioning. Taken together, we believe the presented strategy may serve as a prototypic example for the analysis and utility of employing quantitative-mass-spectrometry-based proteomics to study bacterial physiology.

Sucrose-Induced Proteomic Response and Carbohydrate Utilization of Lactobacillus sakei TMW 1.411 During Dextran Formation

“…replicates) of L. sakei TMW 1.411 were prepared in mMRS as described above (section Bacterial Strain and Growth Conditions) and used to inoculate 4 × 100 ml cultures in mMRS (20 g/L glucose) with a final OD 600 of 0.1. The cultures were grown to the mid-exponential growth phase (pH ~5.0, determined in preliminary experiments), which had given good results in previous experiments (Schott et al, 2017 ), and subsequently distributed to 50 ml sealed tubes each (eight tubes in total). Afterwards, the cultures were pelletized (5,000 × g, 10 min) and washed once in fresh mMRS.…”

“… Overview of the experimental steps for the analysis of sucrose-induced changes in the proteomic profile of L. sakei TMW 1.411. This figure is partly based on Figure 1 of Schott et al ( 2017 ). …”

Technological characterization of vaginal probiotic lactobacilli: resistance to osmotic stress and strains compatibility

“…), expression of a set of proteins or quantitative proteomic (Schott et al . ) or glycine betaine transport (Han et al . ).…”

Adaptation strategies of Pseudomonas protegens SN15‐2 to hyperosmotic growth environment

“…The adaptation mechanism of micro‐organisms to stress environments is an area of current research interest (Velliou et al ; Bustos and Borquez ; Schott et al ). Different micro‐organisms exhibit a variety of adaptive mechanism for the regulation of hyperosmotic conditions including the accumulation of compatible solutes, ion transport and energy metabolism (Sleator and Hill ).…”