STANDARDIZATION OF ENCAPSULATION TECHNIQUE FOR EFFECTIVE PROBIOTIC DELIVERY
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Date
2022-02-11
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COLLEGE OF DAIRY SCIENCE AND TECHNOLOGY MANNUTHY
Abstract
A potentially probiotic lactic acid bacterium was isolated from human breast milk.
Based on biochemical characterization and 16SrRNA sequencing, the isolate was identified as
Pediococcus pentosaceus DM101, the sequence of which is deposited in NCBI with accession
number MK774704. Acid and bile tolerance studies indicated that Pediococcus pentosaceus
DM101 can withstand pH 3.0 and 0.6 % (w/v) bile salt for four hours. The isolate exhibited an
auto-aggregation value of 67.9±0.74% and a cell surface hydrophobicity values of 60.7±0.67%
with xylene signifying the good adhesion potential for the isolate. Inability to cause
haemolysis and liquefy gelatin suggests the possible absence of virulence factors. Antibiogram
revealed the isolate to be resistant to Vancomycin, Methicillin, Bacitracin, and Cephalosporin
group of antibiotics. Congo red assay confirmed the exopolysaccharide (EPS) producing
nature of the isolate. DPPH(2,2-diphenyl picrylhydrazyl) assay of the cell pellet and cell-free
supernatant revealed an IC 50 value of 61.57 and 20.78mg/ml, endorsing its radical
scavenging potential. Based on survival per cent, encapsulation by extrusion was optimized
with 2.0% sodium alginate as gelling agent, 0.1 M CaCl2 as hardening solution and gelling
time 20 minutes. The alginate beads so obtained were characterized in terms of swelling ratio
and dissolution behavior. The alkaline pH 8.0 facilitated the release of bioactive components
by 4h, whereas acidic pH 2.0 did not release the contents even after 8h. When exposed to
simulated gastrointestinal conditions, the survival rates for encapsulated and non-encapsulated
cells were 86.11% and 15.86% respectively. On 60th day of storage, log reduction of viable
cells in encapsulated form at 4°C and -18°C was 1.10 and 3.30 respectively. For the same
time period, corresponding values were 9.17 and 9.30 for non-encapsulated cells. The
observations endorse encapsulation as an effective tool for sustaining viability when exposed
to challenging environment.
Description
Submitted in partial fulfillment of the requirement for the degree of
Master of Technology in
Dairy Microbiology