Lactic acid bacteria (LAB) is defined as live microorganism, when administered in adequate amount confer a health benefit on the host. Raw cow milk was serially diluted, spread plated, sub cultured and pure cultured. Screening of isolates (MK-2 and MK-4) was performed on the basis of zone formation and characterized. The morphological and biochemicalcharacterization was carried out and it was identified as MK-2 (Lactobacillus sp) and MK-4 (Bacillus cereus). Both the organism were screened for Folateproduction by inoculating the microorganism in folate production media. The highest Folatedetected for MK-2 (Lactobacillus), followed by MK-4 (Bacillus cereus) revealed 150 μg/ml of Folate. In strain MK-4 showed 50-60 μg/ml of Folate production. Further study extended for Folateproducing lipase enzyme from MK-2 and MK-4 revealed negative resultfor lipase. Whereas, positive result was obtained for protease enzyme by MK-2 and MK-4. Antimicrobial activity of Lactobacillusagainst selective bacterial pathogens revealed maximum activity was observed against Pseudomonas aeruginosa, E.coli, Staphylococcus aureus, and Shigella flexneri. The antifungal activityrevealed maximum activity against Aspergillus niger and Aspergillus flavus. Probiotic properties of Lactobacillussptested against different concentration of ox bile with MK-2 and MK-4 strain showed resistance against ox bile. The strain MK-4 showed maximum survivability. In another study to assess the probiotic property such as NaCl tolerance, MK-4 strain showed maximum tolerance of6.5%of NaCl. MK-2 showed maximum growth at pH7. The total protein content of folic acid (bacteriocin) produced by MK-2 (Lactobacillus sp) revealed 2.01μg/ml, and the total protein content ofMK-4 showed 1.97 μg/ml protein content respectively. Determination of molecular weight of folic acid for the strain (MK-2 and MK-4) revealed 46 KDa and 30 KDaby SDS PAGE analysis. HPLC analysis was performed to analyze the metabolite present and it was confirmed with prominent peaks with retention time of 1.801 indicate the folic acid metabolite.
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