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Purifica??o e caracteriza??o de uma ?-N-acetillhexosaminadase extra?da do mam?fero marinho Sotalia fluviatilisGomes J?nior, Jos? Edilson 06 December 2006 (has links)
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Previous issue date: 2006-12-06 / This report shows 2232 times purification of a βNAcetylhexosaminidase from hepatic extracts from the sea mammal Sotalia fluviatilis homogenate with final recovery of 8,4%. Sequenced steps were utilized for enzyme purification: ammonium sulfate fractionation, Biogel A 1.5 m, chitin, DEAESepharose and hydroxyapatite chromatographies. The protein molecular mass was estimated in 10 kDa using SDSPAGE and confirmed by MALDITOF. It was found to have an optimal pH of 5.0 and a temperature of 60?C. Using pnitrophenylNAcetylβDglycosaminide apparent Km and Vmax values were of 2.72 mM and 0.572 nmol/mg/min, respectively. The enzyme was inhibited by mercury chloride (HgCl2) and sodium dodecil sulfate (SDS) / Este trabalho mostra a purifica??o de 2232 vezes de uma βNAcetilhexosaminidase obtida a partir dos extratos hep?ticos do mam?fero marinho Sotalia fluviatilis com recupera??o final de 8,4%. Passos seq?enciais foram utilizados para a purifica??o enzim?tica: fracionamento com sulfato de am?nio e as cromatografias de Biogel A 1.5 m, Quitina, DEAESepharose e Hidroxiapatita. A massa molecular prot?ica foi estimada em 10 kDa usando SDSPAGE e confirmada por MALDITOF. Foi encontrado como pH e temperatura ?timos, 5,0 e 60?C, respectivamente. Os valores de Km e Vm?x aparentes foram 2,72 mM e 0,572 nmol/mg/min, sendo utilizado o pnitrofenilNAcetilβDglicosamin?deo como substrato. A enzima foi inibida pelo cloreto de merc?rio (HgCl2) e dodecil sulfato de s?dio (SDS)
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