Browsing by Author "BINDU DEVI"
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ThesisItem Open Access PURIFICATION OF IMPORTANT BIOACTIVE COMPOUNDS SYNTHESIZED BY FUNCTIONALLY PROFICIENT PROBIOTICS AND THEIR GENETIC CHARACTERIZATION(UHF,NAUNI, 2022-02) BINDU DEVI; SHARMA, NIVEDITAABSTRACT The present study was carried out to investigate important bioactive compounds synthesized by functionally proficient probiotics i.e. L. reuteri F8, L. crustorum F11, L. spicheri G2 and L. fermentum L2 isolated from traditional fermented food matrices of Himachal Pradesh. Morphological and genetic characterization using 16S rRNA technique of all four LAB were done. The 4 LAB strains along with reference strain, L. fermentum 214 were further evaluated for their safety assessment and found to be non-hemolytic, nonmucinolytic, non-DNase, non-gelatinase and non-biogenic amine producers. All these were sensitive to antibiotics tested and MIC determined was found to be lower than MIC break points referred by EFSA. All these 4 isolated showed high acidic tolerance, bile salt tolerance, auto-aggregation, co-aggregation and hydrophobicity properties. Therapeutic effects such as antioxidant activity, proteolytic activity, lipid peroxidation inhibition activity, cholesterol lowering, α-glucosidase inhibition and α-amylase inhibition activities were studied. The cumulative probiotic score was calculated and ranged between 77 to 95.83%. Qualitative and quantitative screening of bioactive compounds i.e. lactic acid, EPS and β-galactosidase has been done. L. crustorum F11 has shown maximum lactic acid (3.93 g/L) and EPS (3.50 mg/ml) production and L. fermentum L2 has shown maximum β-galactosidase (33.55 IU/ml) production in quantitative estimation. Percent increase from 16.34-33.64g/L, 11.36 to 32.95 mg/ml and 60.59 to 116.38 IU/ml, respectively was observed after optimization of Lactic acid, EPS and β-galactosidase production using OFAT. Further, using GC-MS technique, metabolite fingerprinting of all four isolates was done and total 20 metabolites were found and recorded that L. reuteri F8 and L. crustorum F11 produces 6 metabolites while L. spicheri G2 has produced 8 metabolites and L. fermentum L2 has produced maximum number (17) of metabolites which are known to exert various beneficial biological effects over the host’s health. Using dendrogram, L. fermentum L2 was selected as the best isolate using SPSS 16 software and used for further studies. Yoghurt was prepared with L. fermentum L2 and reference strain, L. fermentum 214 and physicochemical, microbiological and organoleptic attributes were evaluated. Casein and whey fractions of yogurt were collected and purified using gel filtration chromatography. Fractions with maximum OD at 220 nm were tested for antidiabetic (α-glucosidase and α-amylase inhibition activities) and antioxidative activities. Fatty acid extraction from yoghurt YL2 and Y214 has been done and further subjected for the fatty acid profiling through GC-MS/MS after synthesizing the FAME. Short chain, medium chained and long chain fatty acids ranging from C4:0 to C20:0 were found in yoghurt samples which exert various health benefits.