Proteomic and genetics insights on the response of the bacteriocinogenic Lactobacillus sakei CRL1862 during biofilm formation on stainless steel surface at 10°C.

Some lactic acid bacteria have the ability to form biofilms on food-industry surfaces and this property could be used to control food pathogens colonization. Lactobacillus sakei CR1862 was selected considering its bacteriocinogenic nature and ability to adhere to abiotic surfaces at low temperatures. In this study, the proteome of L. sakei CRL1862 grown either under biofilm on stainless steel surface and planktonic modes of growth at 10°C, was investigated. Using two-dimensional gel electrophoresis, 29 out of 43 statistically significant spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Ten proteins resulted up-regulated whereas 16 were down-regulated during biofilm formation. Differentially expressed proteins were found to belong to carbohydrate, nucleotide, aminoacid and lipid metabolisms as well as translation, peptide hydrolysis, cell envelope/cell wall biosynthesis, adaption to atypical conditions and protein secretion. Some proteins related to carbohydrate and nucleotide metabolisms, translation and peptide degradation were overexpressed whereas those associated to stress conditions were synthesized in lower amounts. It seems that conditions for biofilm development would not imply a stressful environment for L. sakei CRL1862 cells, directing its growth strategy towards glycolytic flux regulation and reinforcing protein synthesis. In addition, L. sakei CRL1862 showed to harbor nine out of ten assayed genes involved in biofilm formation and protein anchoring. By applying qRT-PCR analysis, four of these genes showed to be up regulated, srtA2 being the most remarkable. The results of this study contribute to the knowledge of the physiology of L. sakei CRL1862 growing in biofilm on a characteristic food contact surface. The use of this strain as green biocide preventing L. monocytogenes post-processing contamination on industrial surfaces may be considered.

Antimicrobial activity of whey protein films supplemented with Lactobacillus sakei cell-free supernatant on fresh beef.

The aim of this work was to evaluate the antimicrobial activity of whey protein isolate (WPI) films supplemented with Lactobacillus sakei NRRL B-1917 cell-free supernatant on beef inoculated with Escherichia coli ATCC 25922 or Listeria monocytogenes Scott A; additionally, sensory evaluation was performed on wrapped beef cubes. Supernatant concentrates were obtained from Lb. sakei cultures in MRS broth after centrifugation, filtering, and freeze-drying. Films were prepared with WPI (3% w/w), alginate (0.625% w/w), rehydrated supernatant (18 mg/ml), and glycerol. Films were used to wrap beef cubes inoculated with ≈103 CFU/g E. coli or L. monocytogenes. Sensory evaluation was carried out on grilled beef wrapped or not with the studied antimicrobial films. During refrigerated storage, antimicrobial films reduced 1.4 log10 CFU/g of L. monocytogenes after 120 h, while E. coli decreased 2.3 log10 CFU/g after 36 h. Grilled beef wrapped with antimicrobial film was well accepted by panelists, besides scores evidenced no significant differences (p > 0.05) between wrapped and unwrapped beef.