Genotypic and phenotypic diversity among Lactobacillus plantarum and Lactobacillus pentosus isolated from industrial scale cucumber fermentations.

The Lactobacillus plantarum and Lactobacillus pentosus genotypes existing in industrial-scale cucumber fermentations were defined using rep-PCR-(GTG)5. The ability of each genotype to ferment cucumbers under various conditions was evaluated. Rep-PCR-(GTG)5 was the technique capable of illustrating the most intraspecies discrimination compared to the sequencing of housekeeping genes (recA, dnaK, pheS and rpoA), MLST and RAPD with primers LP1, OPL5, M14 and COC. Ten genotypic clusters were defined for the 199 L. pentosus tested and three for the 17 L. plantarum clones. The ability of the 216 clones genotyped and 37 additional cucumber fermentation isolates, of the same species, to rapidly decrease the pH of cucumber juice medium under various combinations of sodium chloride (0 or 6%), initial pH (4.0 or 5.2) and temperatures (15 or 30 °C) was determined using a fractional factorial screening design. A reduced fermentation ability was observed for the L. plantarum strains as compared to L. pentosus, except for clone 3.2.8, which had a ropy phenotype and aligned to genotypic cluster A. L. pentosus strains belonging to three genotypic clusters (B, D and J) were more efficient in cucumber juice fermentation as compared to most L. plantarum strains. This research identified three genetically diverse L. pentosus strains and one L. plantarum as candidates for starter cultures for commercial cucumber fermentations.

Effect of Brine Acidification on Fermentation Microbiota, Chemistry, and Texture Quality of Cucumbers Fermented in Calcium or Sodium Chloride Brines.

Commercial fermentation for bulk preservation of cucumbers relies on natural microbiota and approximately 1 M sodium chloride (NaCl) brines, resulting in large volumes of high-salt wastewater. An alternative process utilizing 0.1 M calcium chloride (CaCl2 ) as the only salt was developed to eliminate NaCl from fermentation brines for reduced environmental impact. This study determined the effect of brine acidification on the fermentation microbiota and texture quality of cucumbers fermented in CaCl2 brines. Cucumber fermentations were conducted in sealed glass jars for six independent lots of cucumbers in a randomized complete block design with a full-factorial treatment structure for brine acidification (acetic acid, hydrochloric acid, or nonacidified) and brining salt (1 M NaCl or 0.1 M CaCl2 ). Enterobacteriaceae spp. survived longer and were >1 log colony forming units/mL higher in fermenting cucumbers than in brines. Addition of 25 mM acetic acid to fermentation brines (but not the addition of hydrochloric acid at the same pH) reduced Enterobacteriaceae spp. in brines and cucumbers (P < 0.002) during the initiation of fermentation for both brining salts. However, acidification had no effect on texture quality of fermented cucumbers (P = 0.8235). Despite differences in early fermentation microbiota, fermentation of cucumbers in calcium chloride brines under controlled conditions, with or without acidification, resulted in high retention of tissue firmness. These results differ from fermentations in a commercial setting initiated in brines of neutral pH, indicating that production variables, such as air exposure, interact with brining in CaCl2 to negatively affect the texture quality of fermented cucumbers. PRACTICAL APPLICATION: This study examined the effects of initial brine acidification on the course of lactic acid fermentation and resulting texture quality of cucumbers fermented in calcium or sodium salt brines. Fermentation brines containing acetic acid (the acid in vinegar) reduced the pH of the cucumber and the soil-associated Enterobacteriaceae spp. most rapidly, and favored the conversion of sugars to lactic acid. Interestingly, the texture quality was not affected by brine acidification, and all cucumbers fermented in calcium brines in the absence of air retained their firmness during fermentation and bulk storage.

Direct Analysis of Triterpenes from High-Salt Fermented Cucumbers Using Infrared Matrix-Assisted Laser Desorption Electrospray Ionization (IR-MALDESI).

High-salt samples present a challenge to mass spectrometry (MS) analysis, particularly when electrospray ionization (ESI) is used, requiring extensive sample preparation steps such as desalting, extraction, and purification. In this study, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) coupled to a Q Exactive Plus mass spectrometer was used to directly analyze 50-µm thick slices of cucumber fermented and stored in 1 M sodium chloride brine. From the several hundred unique substances observed, three triterpenoid lipids produced by cucumbers, ß-sitosterol, stigmasterol, and lupeol, were putatively identified based on exact mass and selected for structural analysis. The spatial distribution of the lipids were imaged, and the putative assignments were confirmed by tandem mass spectrometry performed directly on the same cucumber, demonstrating the capacity of the technique to deliver confident identifications from highly complex samples in molar concentrations of salt without the need for sample preparation. Graphical Abstract ᅟ.