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Stabilisation of carboxypeptidases in acid extracts of ovine pancreas
Institution:1. Faculty of Health and Sport Sciences, University of Tsukuba, Tsukuba, Japan;2. Department of Sport Science, Japan Institute of Sport Sciences, Tokyo, Japan;3. Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan
Abstract:Carboxypeptidases A (CPA) and B (CPB) are pancreatic exopeptidases that are very sensitive to environmental stresses such as freezing, drying, temperature and pH. Traditional low pH methods of extracting pancreatic enzymes destroy carboxypeptidase activity. The use of sucrose as a stabilising agent prevents pH induced denaturation of carboxypeptidases during their extraction. Greater than 70% carboxypeptidase A activity can be retained at pHs as low as 3.5 using 0.5 M sucrose. In addition extraction at low pH ensures good trypsin and chymotrypsin recovery while elastase recovery is improved due to the stabilising effect of sucrose. Further fractionation of the extract produces preparations enriched in endopeptidases or exopeptidase suitable for use in protein hydrolysis applications.
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