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Characterization and Potent Application of Pleurotus floridanus Trypsin Inhibitor (PfTI)

Natural Product Sciences 2020년 26권 3호 p.207 ~ 213
Pannippara Manzur Ali, Kesav Sapna, Raghavan Rekha Mol Kollakal Naduvil, Mathew Abraham, Bhat Sarita Ganapathy, Kozhiyil Elyas Kothanan,
소속 상세정보
 ( Pannippara Manzur Ali ) - Cochin University of Science and Technology Department of Biotechnology
 ( Kesav Sapna ) - Cochin University of Science and Technology Department of Biotechnology
 ( Raghavan Rekha Mol Kollakal Naduvil ) - Cochin University of Science and Technology Department of Biotechnology
 ( Mathew Abraham ) - Cochin University of Science and Technology Department of Biotechnology
 ( Bhat Sarita Ganapathy ) - Cochin University of Science and Technology Department of Biotechnology
 ( Kozhiyil Elyas Kothanan ) - Calicut University Department of Biotechnology

Abstract


Characterization and in vitro inhibition studies of protease inhibitor from the mushroom Pleurotus floridanus (PfTI) towards the pest Papilio demoleus is studied. The addition of 1 mM Mn2+, Na2+, Ba2+ and Ni 2+ enhanced the PfTI activity. The ICP-atomic emission spectrum showed the presence of Ca2+, Mg2+ and Zn2+ in the PfTI. Surfactants SDS and CTAB at a concentration of 1% reduced the PfTI activity whereas, the nonionic detergents Triton X and Tween 80 increased the activity. The inhibitory activity gradually decreased with increase in concentration of DMSO and H2O2. The activity was increased by dithiothreitol up to a concentration of 80 μM and inactivated at 140 μM. The activity of PMSF modified PfTI was drastically reduced to 0.234 U/mL at 4 mM concentration and similar results were obtained for modification of cysteine by N-Ethylmaleimide at slightly higher concentrations. The complex of trypsin and PfTI showed complete loss in fluorescence intensity at 343 nm compared with control. In vitro inhibition studies of PfTI with midgut proteases isolated from citrus pest P. demoleus with protease activity of 1.236 U was decreased to 0.613 U by 50 μL (0.1 mg/mL) of the inhibitor. Inhibitor was stable up to 0.04 M concentration of HCl.

키워드

Trypsin inhibitor; Pleurotus floridanus; Characterization

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