Methods for maintaining and using lactic acid bacteria starter cultures for commercial cucumber fermentation brined with reduced salt

Authors: Perez Diaz, Ilenys; Santos, Amanda; Page, Clinton; SANTOS, FERNANDA - North Carolina State University; ARROYO-LOPEZ, FRANCISCO NOE - Instituto De La Grasa

Submitted to: Journal of Food Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/23/2026
Publication Date: 8/21/2026
Citation: Perez Diaz, I.M., Santos, A., Page, C.A., Santos, F., Arroyo-Lopez, F. 2026. Methods for maintaining and using lactic acid bacteria starter cultures for commercial cucumber fermentation brined with reduced salt. Journal of Food Science. 91. https://doi.org/10.1111/1750-3841.71363.
DOI: https://doi.org/10.1111/1750-3841.71363

Interpretive Summary: US pickle processors need vegetable fermentation technology with low salt to expand production capacity while respecting the environment and making healthier products. Implementation of commercial cucumber fermentations with reduced salt demands the utilization of starter cultures to guide fermentations to the endpoint, assure acid-production, and prevent processing defects. We evaluated the long-term storage of selected lactic acid bacteria starter cultures in Microbanks that were stored in a household freezer. While starter cultures for dairy products are readily available worldwide, the plant-derived counterparts proficient in pickling are scarce. The relatively small volume of starter cultures needed by the pickling industry in the U. S. A., relative to the dairy industry, is unattractive for their profitable commercialization. Currently, in the U. S. A., there is a handful of companies that manufacture starter cultures for pickling. An example is the Picallili Vpro that consist of Lactiplantibacillus pentosus LA0445 proficient in the fermentation of cucumbers to pH 3.3. It was determined that starter cultures for pickling can be maintained in Microbanks in a household freezer for up to 2 years. A method compatible with tank yard operations was developed for processors to make sufficient volumes of starter cultures for pickling. Such method creates opportunities for processors to generate starter cultures in-house for commercial scale cucumber fermentations using functional strains of lactic acid bacteria isolated from such habitat.

Technical Abstract: We describe the long-term storage of lactic acid bacteria (LAB) in Microbanks at -20 °C and a method for their utilization as starter cultures in commercial cucumber fermentation. The Microbank manufacturer’s recommendations apply for the preservation of Lactococcus lactis, and Levilactobacillus brevis strains for up to 1 year, while that of Pediococcus pentosaceous and the Lactiplantibacillus is possible for up to 2 years. LAB known to produce exopolysaccharides, such as Lactiplantibacillus, remain viable in frozen Microbanks for more than 2 years. The preserved LAB cultures were resuscitated in cucumber, carrot, or sweetpotato juice after 24 to 30 h of incubation at ambient temperature. Cucumber Fermentation Medium (CFM), used for the pre-adaptation of the starter cultures to 342 mM (2 %) sodium chloride (NaCl), sustained the growth of the preserved LAB to maximum cell densities achievable in 12 to 24 h. The pre-adaptation of the cultures in 100 mL of CFM supported quick acid production, but inoculation of up to 10 L of CFM with a 10 µL loop full of cells sufficed for a complete fermentation and production of biomass. Cultures of Lc. lactis and Lp. plantarum reached maximum cell densities in CFM in 12 h, while Lp. pentosus presented a 2 h delay (14 h). Cultures of Lev. brevis and Pediococcus pentosaceous need a 16 h incubation to achieve maximum cell densities. Acid production by the LAB resuscitated in CFM supplemented with 342 mM NaCl, 18 mM calcium chloride, and 18 mM calcium hydroxide reduced the pH to 5.8 ± 0.3 by the time they reached maximum cell densities. Observations made led to a method for the maintenance of LAB starter cultures in Microbanks using a standard household freezer for storage, and a process for their use in preparing inocula for commercial cucumber fermentations that is compatible with tank yard operations.