Abstract:

In this study, we optimized the conditions for enzymatic extraction of polysaccharides from pomegranate seeds
with a mixed enzyme preparation consisting of pectinase, cellulase and mannase. The optimization of the mixing ratio of the
three enzymes was performed by the combined use of one-factor-at-a-time method and D-optimal mixture design. Thereafter,
the enzymatic extraction conditions were optimized by response surface methodology using Box-Behnken design. The yield
of polysaccharides was taken as response for both optimizations. It was shown that the optimal combination of enzymes
were 24.2% pectinase, 67.2% cellulase and 8.6% mannose and that the optimal extraction conditions were determined as
follows: liquid/solid ratio, 20:1 (mL/g); initial pH, 4.5; hydrolysis temperature, 48.5 ℃; and hydrolysis time, 287 min. Under
these conditions, the yield of polysaccharides was 2.83%.

Key words: pomegranate seeds, polysaccharide, enzymatic extraction with mixed enzymes, D-optimal mixture design, response surface analysis, extraction process