Mixed culture fermentation using Lactobacillus delbrueckii and novel Lactiplantibacillus plantarum strains isolated from traditional chhurpi from Sikkim Himalayan region for potential probiotic and improved functional properties

The aim of the study was to isolate potential probiotic lactic acid bacteria (LAB) strains from the traditional fermented milk products, Chhurpi of the Sikkim Himalayan region and to prepare curd using mixed culture fermentation. A preliminary probiotic assessment of 110 LAB isolates including antimicrobial activity, bile and acid tolerance, surface hydrophobicity, auto-aggregation and co-aggregation, and adhesion to Caco-2 cells, was performed. Lactiplantibacillus plantarum strains N4, CHDE2, CHDE4, and SS22 demonstrated better viability in bile (0.3%) and acid (pH 2.5), adherence with hydrophobicity (13.93–92.85%), auto-aggregation (72.44–89.74%), adhesion to Caco-2 cells (45–73.61%) and antimicrobial activity against food borne pathogens. These strains were used for mixed culture fermentation with a previously screened curd forming strain Lactobacillus delbrueckii WS4. Curd produced using mixed culture fermentation of Lb. delbrueckii WS4 and the selected Lp. plantarum strains had a relatively higher acid production (pH 3.97–4.01), protein hydrolysis, and antioxidant activity. Further, mixed culture fermentation also resulted in the increase of HOCl scavenging activity (12.22–32.96%), myeloperoxidase (MPO) inhibition activity (22.38–42.94%) and angiotensin converting enzyme (ACE) inhibition (15.61–33.03%) as compared to that of WS4 fermentation (P < 0.05). This is the first report on identification of potential probiotic strains from the traditional fermented milk products of Sikkim and application in mixed culture fermentation with enhanced multifunctional potential.