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Biochemical characterisation of a Kunitz-type inhibitor from Tamarindus indica L. seeds and its efficacy in reducing plasma leptin in an experimental model of obesity
Authors:Amanda Fernandes de Medeiros  Izael de Sousa Costa  Fabiana Maria Coimbra de Carvalho  Sumika Kiyota  Beatriz Blenda Pinheiro de Souza  Daniel Nogoceke Sifuentes
Institution:1. Postgraduate Biochemistry Program, Biosciences Center, Federal University of Rio Grande do Norte, Natal, Brazil;2. Laboratory of Protein and Peptide Biochemistry, CPDSA, Biological Institute, S?o Paulo, Brazil;3. Postgraduate Biological Molecular, Institute of Biological Sciences, University of Brasília, Distrito Federal, Brasília, Brazil;4. Embrapa Genetic Resources and Biotechnology, Embrapa, Distrito Federal, Brasília, Brazil;5. Embrapa Genetic Resources and Biotechnology, Embrapa, Distrito Federal, Brasília, Brazil
Abstract:A trypsin inhibitor isolated from tamarind seed (TTI) has satietogenic effects in animals, increasing the cholecystokinin (CCK) in eutrophy and reducing leptin in obesity. We purified TTI (pTTI), characterised, and observed its effect upon CCK and leptin in obese Wistar rats. By HPLC, and after amplification of resolution, two protein fractions were observed: Fr1 and Fr2, with average mass of M?+?14H]+?=?19,594,690?Da and M?+?13H]+?=?19,578,266?Da, respectively. The protein fractions showed 54 and 53 amino acid residues with the same sequence. pTTI presented resistance to temperature and pH variations; IC50 was 2.7?×?10?10?mol.L?1 and Ki was 2.9?×?10?11?mol.L?1. The 2-DE revealed spots with isoelectric points between pH 5 and 6, and one near pH 8. pTTI action on leptin decrease was confirmed. We conclude that pTTI is a Kunitz trypsin inhibitor with possible biotechnological health-related application.
Keywords:Tamarind  antitryptic  Wistar rats  CCK
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