Purification and characterization of aminopeptidase N from chicken intestine with potential application in debittering |
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| Institution: | 1. Department of Biotechnology, University of Pune, Ganeshkhind, Pune-411007, India;2. Food Technology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085, India;1. School of Food Science and Technology, Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia;2. Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia;1. College of Chemistry, Chemical Engineering and Food Safety, Bohai University, Jinzhou, 121013, PR China;2. Department of Food Science and Technology, University of Georgia, GA, 30223-1791, USA;1. Post Graduate Program in Food Science and Technology, Department of Food Engineering, Technology Centre, Federal University of Paraiba, Campus I, 58051-900, João Pessoa, Paraíba, Brazil;2. Food Technology Institute (ITAL). Science and Food Quality Center, 13070-179, Campinas, São Paulo, Brazil;1. Food Technology and Nutrition Laboratory, Abdelhamid Ibn Badis University, Mostaganem, Algeria;2. Higher School of Agronomy, Mostaganem, Algeria;3. Structure, Development and Application of Molecular Materials Laboratory, Abdelhamid Ibn Badis University, Algeria;4. Food Technology Program Laboratory, University of Malaysia, Terengganu, Malaysia;5. Department of Plant Protection, Faculty of Agriculture, Damascus University, Syrian Arab Republic |
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| Abstract: | An aminopeptidase with broad substrate specificity was purified to homogeneity (123.7-fold) with a yield of 3.43% from chicken (Gallus gallus) intestine using a combination of chromatographic separation strategies. The enzyme was identified as alanyl aminopeptidase or aminopeptidase N (APN) by Peptide Mass Fingerprinting. The molecular weight of the enzyme was estimated to be ∼180 kDa by SDS-PAGE and gel filtration chromatography. The enzyme was found to be a glycoprotein, having 40% sugar residue and a molecular mass of 108 kDa after deglycosylation. The enzymatic activity was optimal at 60 °C and pH 6.0. The enzyme preferentially hydrolyzed Leu-β-NA (Km = 0.1 mM) followed by Ala, Phe, Tyr and Gly at N-terminal. The enzyme activity was completely inhibited by 1,10 phenanthroline (1 mM) and bestatin (1 mM) confirming it as a metalloprotease. Potential of this enzyme in combination with other endoproteases for the production of debittered protein hydrolysates has been discussed. |
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