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Preparation and Characterization of a Trypsin Inhibitor from G lycine max ( L .) merr
Author(s) -
Wang Shaoyun,
Shao Biao,
Cai Xixi,
Rao Pingfan,
Deng Zeyuan,
Xie Mingyong
Publication year - 2014
Publication title -
journal of food processing and preservation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.511
H-Index - 48
eISSN - 1745-4549
pISSN - 0145-8892
DOI - 10.1111/jfpp.12182
Subject(s) - pythium aphanidermatum , ammonium sulfate precipitation , trypsin , isoelectric point , botrytis cinerea , molecular mass , biochemistry , glycine , chemistry , trypsin inhibitor , isoelectric focusing , chromatography , fusarium oxysporum , affinity chromatography , amino acid , biology , enzyme , size exclusion chromatography , botany , biological pest control
A trypsin inhibitor with thermal and pH stability, designated M ercine , was prepared from seeds of G lycine max ( L .) merr . The preparation procedure involved ammonium sulfate precipitation, ion‐exchange chromatography on CM ‐ S ephadex C ‐50, affinity chromatography on A ffi‐gel blue gel. The 20 N ‐terminal amino acid sequences were determined to be DEYSKPCCDLCMCTRRMPPQ , demonstrating highly homologies with the sequence of B owman– B irk type trypsin inhibitors. The molecular mass and isoelectric point of the inhibitor were estimated by SDS ‐ PAGE and isoelectric focusing to be 17.9 kD and 4.6, respectively. Trypsin could be completely inhibited by M ercine when the molar ratio was 8.0. The inhibitory activity of M ercine was unaffected after exposure to temperatures up to 80C, or within the pH range 2–12. Besides inhibiting trypsin‐chymotrypsin, the inhibitor M ercine demonstrated additional antifungal activity toward the species of A lternaria alternate, F usarium oxysporum , P ythium aphanidermatum , P hysalospora piricola and B otrytis cinerea . Practical Applications We here report, for the first time, not only the trypsin inhibitor's preparation, but also its N ‐terminal amino acid sequence and antifungal activity against a series of phytopathogenic fungi. The aforementioned activities shown by trypsin inhibitor, provide further evidence for the potential significance in agriculture.