Isolation and Purification of Trypsin Inhibitors from the Seeds of Abelmoschus moschatus L.

Abstract

Four trypsin inhibitors, AMTI-I, AMTI-II, AMTI-III, and AMTI-IV, have been isolated and purified to homogeneity from the seeds of Abelmoschus moschatus following ammonium sulphate fractionation, DEAE-cellulose ion exchange chromatography and gel permeation on Sephadex G-100, and their molecular weights were determined to be 22.4, 21.2, 20.8 and 20.2 kDa respectively by SDS-PAGE. While all the four inhibitors were very active against bovine trypsin, two of them (AMTI-III and AMTI-IV) showed moderate activity towards bovine chymotrypsin. AMTI-I and AMTI-II were found to be glycoproteins with neutral sugar content of 2.8 and 4 %, respectively, and all the four inhibitors were devoid of free sulphhydryl groups. The inhibitors were quite stable up to 80 °C for 10 min and were not affected at alkaline as well as acidic conditions tested. Treating them with 8 M urea and 1 % SDS for 24 h at room temperature did not result in any loss of their antitryptic activities. However, they lost considerable antitryptic activity when treated with 6 M guanidine hydrochloride. Activities of the inhibitors were unaffected even after their reduction with DTT suggesting that disulphide bonds are not needed for their inhibitory activities.

Application and bioactive properties of CaTI, a trypsin inhibitor from Capsicum annuum seeds: membrane permeabilization, oxidative stress and intracellular target in phytopathogenic fungi cells

“…isolated different proteinase inhibitors from C. chinense seeds and found that these could inhibit the growth of the yeasts S. cerevisiae , C. albicans , C. tropicalis , Pichia membranifaciens and Kluyveromyces maxiannus , which cause morphological changes in these cells. Dokka and Davuluri identified in Abelmoschus moschatus seeds a trypsin inhibitor with activity against various fungi such as C. albicans , C. tropicalis , Aspergillus flavus , S. cerevisiae , C. glaba and A. niger . In another study, Chilosi et al .…”

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“…AMTI-II also found to moderately affects the growth of fungal species, C. albicans, Candida tropicalis, Aspergillus flavus, Saccharomyces cerevisiae, Candida glabrata and Aspergillus niger. [16] …”

“…It is concluded that the AMTI-I and AMTI-II exhibit stable and potent hemagglutinating property against human and animal erythrocytes and may be useful in agricultural field for the development of insect-resistant transgenic crops. [23] Anti-ageing Property A cosmetic preparation containing A. moschatus seed extract was developed and studied for its effect on skin fibroblast in vitro and in vivo. Results of in vitro experiment showed that A. moschatus seed extract exhibits heparan sulfatelike properties and dose-dependently protects FGF-2 from thermal degradation.…”

Characterization of Monoheaded Trypsin Inhibitors From the Seeds of Abelmoschus Moschatus L.

“…Two trypsin inhibitors (Abelmoschus moschatus trypsin inhibitor-I [AMTI-I] and AMTI-II) isolated and purified from the seeds of A. moschatus have been found to be homogenous by the criteria of native PAGE and gel filtration [23]. Investigations on the specificity of protease inhibitors are important for understanding their physiological role, control mechanisms involved in the regulation of proteolysis in biological systems and mode of action.…”