Purification and characterization of a new trypsin inhibitor from Dimorphandra mollis seeds |
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| Authors: | Mello G C Oliva M L Sumikawa J T Machado O L Marangoni S Novello J C Macedo M L |
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| Institution: | (1) Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil;(2) Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campo dos Goytacazes, RJ, Brazil;(3) Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil;(4) Departamento de Ciências Naturais, Universidade Federal de Mato Grosso do Sul (CEUL/UFMS), CP. 549, 79603-011 Três Lagoas, MS, Brazil |
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| Abstract: | A second trypsin inhibitor (DMTI-II) was purified from the seed of Dimorphandra mollis (Leguminosae-Mimosoideae) by ammonium sulfate precipitation (30–60%), gel filtration, and ion-exchange and affinity chromatography. A molecular weight of 23 kDa was estimated by gel filtration on a Superdex 75 column SDS-PAGE under reduced conditions showed that DMTI-II consisted of a single polypeptide chain, although isoelectric focusing revealed the presence of three isoforms. The dissociation constant of 1.7 × 10–9 M with bovine trypsin indicated a high affinity between the inhibitor and this enzyme. The inhibitory activity was stable over a wide pH range and in the presence of DTT. The N-terminal sequence of DMTI-II showed a high degree of homology with other Kunitz-type inhibitors. |
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| Keywords: | Dimorphandra mollis Mimosoideae trypsin inhibitor N-terminal sequence Kunitz family |
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