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Isolation and partial purification of beta-galactosidases from cotyledons of two cowpea cultivars

Isolamento e purificação parcial de beta-galactosidases de cotilédones de dois cultivares de feijão-de-corda

Three isoforms of beta-galactosidases were isolated and partially purified from the cotyledons of quiescent seeds of Vita 3 and Vita 5 cowpea [Vigna unguiculata (L.) Walp.] cultivars differing in water and salt stress tolerance. The purification procedure consisted of ammonium sulfate fractionation, acid precipitation, ion exchange chromatography through DEAE-sephadex and affinity chromatography through Lactosyl-sepharose columns. The three isoforms isolated from the two cultivars showed the same chromatographic patterns, same optimum of temperature for enzyme activity assay (60ºC), identical thermal stability up to 50°C, and similar pH optima (3-4). However, they differed from each other in sensitivity towards metal ions and certain chemical agents presents in the assay medium. The results have shown that the observed differences in beta-galactosidases from the cotyledons of quiescent seeds were not sufficient to relate them to stress tolerance.

cotyledons; isozymes; Vigna unguiculata; Vita 3 and Vita 5 cultivars; salt tolerance


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