Chandrasekaran, M; Manzur Ali, P P; Rekha Mol, K R; Sapna, K; Sarita,G Bhat(Springer, March 11, 2014)
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Abstract:
Protease inhibitors can be versatile tools mainly in the fields of medicine, agriculture and
food preservative applications. Fungi have been recognized as sources of protease inhibitors,
although there are only few such reports on mushrooms. This work reports the purification and
characterization of a trypsin inhibitor from the fruiting body of edible mushroom Pleurotus
floridanus (PfTI) and its effect on the activity of microbial proteases. The protease inhibitor was
purified up to 35-fold by DEAE-Sepharose ion exchange column, trypsin-Sepharose column and
Sephadex G100 column. The isoelectric point of the inhibitor was 4.4, and its molecular mass was
calculated as 37 kDa by SDS-PAGE and 38.3 kDa by MALDI-TOF. Inhibitory activity confirmation
was by dot-blot analysis and zymographic activity staining. The specificity of the inhibitor
toward trypsin was with Ki of 1.043×10−10 M. The inhibitor was thermostable up to 90 °C with
maximal stability at 30 °C, active over a pH range of 4–10 against proteases from Aspergillus
oryzae, Bacillus licheniformis, Bacillus sp. and Bacillus amyloliquefaciens. Results indicate the
possibility of utilization of protease inhibitor from P. floridanus against serine proteases
Description:
Appl Biochem Biotechnol (2014) 173:167–178
DOI 10.1007/s12010-014-0826-1