Thermostable serine protease inhibitor from Death cap (Amanita phalloides)
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Abstract
Background: Protease inhibitor plays an important role in many biological processes in an organism, its selective binding toward protease potentially tuning down some specific biological processes such as enzymatic catalysis regulation, protein signaling as well as protein clearance in order to accomplish homeostasis. Several protein-based protease inhibitors have been isolated and identified, the majority are directed toward serine protease.
Objectives: This study aimed to find a potential protease inhibitor from the local Northern Thailand Death cap (Amanita phalloides) together with biochemical characterization of its general properties.
Materials and methods: Death cap extract collected from Phayao Province, Thailand was initially performed trypsin inhibitory activity assay using BSA and alzoalbumin as substrates. Detection of its inhibition activity was assessed by SDS-PAGE and spectrophotometry. Additionally, molecular size was observed by filtering the extract through 3 kDa molecular cut off membrane. Finally, hydrophobic property was verified by passing the filtrate through phenyl sepharose column chromatography.
Results: Death cap contained serine protease inhibitory activity. Molecular size of inhibitor was suspected to be less than 3 kDa. Hydrophobic property of this inhibitor was observed. Interestingly, its inhibitory property retained after heat inactivation at 100 ᵒC for 10 min.
Conclusion: A novel heat-tolerant inhibitor from the water extract of Death cap was characterized to be a small peptide with hydrophobic property, which could be used as a new peptide protease inhibitor targeting to serine protease that benefit for agricultural and medical field.
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Personal views expressed by the contributors in their articles are not necessarily those of the Journal of Associated Medical Sciences, Faculty of Associated Medical Sciences, Chiang Mai University.
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