CHARACTERIZATION OF LACTIC ACID BACTERIA AS PROBIOTICS AND TO ASSESS THEIR FUNCTIONAL PROFICIENCY IN A FERMENTED BEVERAGE
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Date
2023-11-26
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UHF Nauni
Abstract
The present investigation was carried out to evaluate lactic acid-producing isolates for their
application in the preparation of nutritionally enriched functional food. Total, 3 in-house lactic acid bacterial strains
isolated from 3 different food matrices viz. Gundruk, Dhaheli, and corn molasses (reference strain) were selected for
the evaluation of their therapeutic potential as probiotics. Safety assessment of the isolates were done by evaluating
hemolytic, DNase, gelatinase activities, and antibiotic susceptibility. All three isolates exhibited non-hemolytic, non-
DNase, and non-gelatinase activities and were highly sensitive to all the antibiotics, thereby proving overall safe
status. These screened isolates were further assessed for their probiotic potential viz. acidity tolerance, bile tolerance,
α-amylase and GABA activity. All three screened isolates were highly acidity tolerant with 92%, 78% respectively at
a low pH i.e. 3.0 after 3 h of incubation These three isolates were also able to resist high bile salt concentration i.e.
86%, 89 % survival rate at 1.5% bile salt concentration after 4 h. . The amylase activity of all three isolates i.e. L.
spicheri G2, E. lactis Dh2. and L. delbrueckii 2025 had exhibited clear zones and were designated as amylaseproducing
strains. Gamma amino butyric acid (GABA) activity was analyzed and L. spicheri G2 and L. delbrueckii
had showed higher GABA production in terms bright yellow colored colonies as compared to E. lactis Dh2.
Qualitative and quantitative analysis of selected probiotics isolates was done. In Qualitative analysis, the colour
change from yellow to pink indicated the presence of lactic acid. In the case of quantitative analysis, the maximum
titratable acidity and lactic acid were observed in G2 i.e. 2.88% and 4.17g/L as a compared to Dh2 and 2025 i.e.
1.44% and 3.72 g/L respectively. Out of three isolates, two were selected on the basis of higher lactic acid production.
Optimization of COFAT for LA production of selected probiotic iaolates i.e G2, Dh2, and 2025 was done where strain
L. spicheri G2 showed production of LA on different process parameters i.e carbon source concentration: (5.1g/L),
pH: (4.72g/L), inoculum size (5.44g/L), temperature: (4.40g/L) ,incubation time: (8.12g/L) and strain L. delbrueckii
2025 showed production of LA on different process parameters i.e carbon source concentration: (4.93g/L), pH:
(4.17g/L), inoculum size(4.91g/L), temperature:(5.38g/L), incubation time: (6.52g/L). After optimization through
OFAT, the percent increment of lactic acid for LA production of probiotic strain i.e L. spicheri G2 with initial LA was
22.30% and increased to 59.21% where L. delbrueckii 2025 with initial LA was 21.02 and increased to 32.25%. RSM
overall increase in LA production by L. spicheri G2 was observed to be 8.05mg/ml with optimized conditions i.e. 48 h
incubation time, 35°C, 6.0 pH, 2% carbon concentration,. In the case of L. delbrueckii 2025 with 6.52g/L LA
production under optimized conditions were 48 h incubation time, 35°C temperature, 6.0 pH, 2% carbon
concentration. Depending upon these better probiotic potential L. spicheri G2 and L. delbrueckii 2025 were selected
for further study been used to prepare probiotic enriched functional probiotic red rice beverage with or without peach
pulp. Rice beverage fermented with lactic acid producing probiotic bacteria had overall enhanced its health promoting
properties besides nutritional profile, thus designating it as a functional drink having good taste, aroma and appearing
color. In total, increased nutritional, therapeutic and desirable phyisco-chemical characteristics make it a functional
proficient fermented beverage rich in lactic acid producing probiotics.
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CHARACTERIZATION OF LACTIC ACID BACTERIA AS PROBIOTICS AND TO ASSESS THEIR FUNCTIONAL PROFICIENCY IN A FERMENTED BEVERAGE