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Techno-functional differentiation of two vitamin B12 producing Lactobacillus plantarum strains: an elucidation for diverse future use

Bhushan, Bharat, Tomar, S. K., Chauhan, Arun
Applied microbiology and biotechnology 2017 v.101 no.2 pp. 697-709
Lactobacillus plantarum, Lactobacillus reuteri, bile, biofortification, fermentation, functional foods, gastrointestinal system, hydrophobicity, immunoassays, nucleotide sequences, pathogens, phylogeny, polymerase chain reaction, probiotics, soymilk, structural genes, vitamin B12
An appropriate selection of Lactobacillus strain (probiotic/starter/functional) on the basis of its techno-functional characteristics is required before developing a novel fermented functional food. We compared vitamin B₁₂ (B₁₂, cobalamin) producing Lactobacillus plantarum isolates, BHM10 and BCF20, for functional (vitamin over-production, genomic insight to B₁₂ structural genes, and probiotic attributes) and technological [milks (skim and soy) fermentation and B₁₂ bio-fortification] characteristics. Addition of B₁₂ precursors (5-amonolevulinate and dimethylbenzimidazole) to cobalamin-free fermentation medium increased vitamin production in BHM10, BCF20, and DSM20016 (a positive standard) by 3.4-, 4.4-, and 3.86-folds, respectively. Three important B₁₂ structural genes were detected in L. plantarum species (strains BHM10 and BCF20) by PCR for the first time. The gene sequences were submitted to NCBI GenBank and found phylogenetically closer to respective sequences in B₁₂ producing Lactobacillus reuteri strains. During comparative probiotic testing, BCF20 showed significantly higher (p < 0.05 to p < 0.001) gastrointestinal tolerance and cell surface hydrophobicity (p < 0.05) than BHM10. Moreover, only BCF20 was found positive for BSH activity and also exhibited comparatively better antagonistic potential against potent pathogens. Conversely, high acid and bile susceptible strain BHM10 displayed significantly higher soy milk fermentation and resultant B₁₂ bio-fortification abilities during technological testing. Two B₁₂ quantification techniques, UFLC and competitive immunoassay, confirmed the in vitro and in situ bio-production of bio-available form of B₁₂ after BHM10 fermentation. Conclusively, techno-functional differentiation of two B₁₂ producing strains elucidates their diverse future use; BCF20 either for B₁₂ over-production (in vitro) or as a probiotic candidate, while BHM10 for cobalamin bio-fortification (in situ) in soy milk.