Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/58455
Type: Artigo de periódico
Title: Purification and characterization of multiple forms of soybean seed acid phosphatases
Author: Ferreira, CV
Granjeiro, JM
Taga, EM
Aoyama, H
Abstract: Four isoforms of acid phosphatase (EC 3.1.3.2), API, AP2, AP3A and AP3B, have been detected and partially purified from soybean seed (Glycine max) through DEAE- and SP-Sephadex chromatographies. Specific activity values of 822, 163, 14 and 66 nkat.mg(-1) were obtained for API (903-fold purification), AP2 (180-fold), AP3A (15-fold), and AP3B (73-fold), respectively, using p-nitrophenylphosphate as substrate. Relative native molecular mass values for API, AP2, AP3A and AP3B, determined by gel filtration on calibrated SW-300 Waters Protein Glass column, were found to be 51 000, 58 000, 52 000 and 30 000, respectively. All four acid phosphatase isoforms presented a carbohydrate moiety in their structures and revealed only single phosphatase activity bands following nondenaturing polyacrylamide gel electrophoresis, at pH 8.3. AP1 and AP2 exhibited greater substrate specificity than AP3A and AP3B. The K-m values were determined for p-nitrophenylphosphate, tyrosine-phosphate and inorganic pyrophosphate, at pH 5.0 and 37 degrees C. The acid phosphatases presented the following apparent K-m values: API (pNPP - 0.49, PPi - 0.21 and TyrP - 1.14 mM); AP2 (pNPP - 0.38, PPi - 1.33 and TyrP - 1.14 mM); AP3A (pNPP1 - 0.20, PPi - 0.16 and TyrP - 0.19 mM) and AP3B (pNPP - 0.086, PPI 0.17 and TyrP 0.17 mM). All four isoforms were inhibited by inorganic phosphate, fluoride, vanadate, molybdate, Cu2+ and Zn2+. The soybean seed acid phosphatases did not catalyze the transphosphorylation reaction since no stimulation was observed with inorganic phosphate accepters, such as glycerol, methanol and ethanol. (C) Elsevier, Paris.
Subject: soybean seed
acid phosphatase
purification
kinetic
Glycine max
Country: França
Editor: Gauthier-villars/editions Elsevier
Rights: fechado
Identifier DOI: 10.1016/S0981-9428(98)80173-3
Date Issue: 1998
Appears in Collections:Unicamp - Artigos e Outros Documentos

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