The esterase activity was determined
spectrophotometrically by measuring the hydrolysis of 4-nitrophenyl
butyrate (pNPB) as a substrate.26 (link) The pNPB
dissolved in acetonitrile (50 mM) was added to 200 μL of Na
phosphate buffer (100 mM, pH 7.5) with 0.5% (v/v) Triton X-100 to
0.5 mM pNPB as the final concentration; 10 μL of crude extract
was used as an enzyme source. The enzymatic reaction was carried out
at 25 °C for 15 min, and the release of pNP was measured at 405
nm. Enzyme activity was calculated using the extinction coefficient
of pNP corresponding to 18.5 mM–1 cm–1. One Unit is defined as the amount of the enzyme that catalyzes
the conversion of one micromole of substrate per minute under the
specified conditions of the assay method and normalized by grams of
the fermented substrate (U/g). The statistical correlation between
groups was analyzed by Student’s t-test.
spectrophotometrically by measuring the hydrolysis of 4-nitrophenyl
butyrate (pNPB) as a substrate.26 (link) The pNPB
dissolved in acetonitrile (50 mM) was added to 200 μL of Na
phosphate buffer (100 mM, pH 7.5) with 0.5% (v/v) Triton X-100 to
0.5 mM pNPB as the final concentration; 10 μL of crude extract
was used as an enzyme source. The enzymatic reaction was carried out
at 25 °C for 15 min, and the release of pNP was measured at 405
nm. Enzyme activity was calculated using the extinction coefficient
of pNP corresponding to 18.5 mM–1 cm–1. One Unit is defined as the amount of the enzyme that catalyzes
the conversion of one micromole of substrate per minute under the
specified conditions of the assay method and normalized by grams of
the fermented substrate (U/g). The statistical correlation between
groups was analyzed by Student’s t-test.
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