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Characterization and biotechnological application of an acid alpha-galactosidase from Tachigali multijuga Benth. seeds
Authors:Fialho Lílian da Silva  Guimarães Valéria Monteze  Callegari Carina Marin  Reis Angélica Pataro  Barbosa Daianny Silveira  Borges Eduardo Euclydes de Lima  Moreira Maurilio Alves  de Rezende Sebastião Tavares
Institution:Departamento de Bioquímica e Biologia Molecular - BIOAGRO, Universidade Federal de Vi?osa, Vi?osa, MG 36570-000, Brazil.
Abstract:Tachigali multijuga Benth. seeds were found to contain protein (364 mg g(-1)dwt), lipids (24 mg g(-1)dwt), ash (35 mg g(-1)dwt), and carbohydrates (577 mg g(-1)dwt). Sucrose, raffinose, and stachyose concentrations were 8.3, 3.0, and 11.6 mg g(-1)dwt, respectively. alpha-Galactosidase activity increased during seed germination and reached a maximum level at 108 h after seed imbibition. The alpha-galactosidase purified from germinating seeds had an M(r) of 38,000 and maximal activity at pH 5.0-5.5 and 50 degrees C. The enzyme was stable at 35 degrees C and 40 degrees C, but lost 79% of its activity after 30 min at 50 degrees C. The activation energy (E(a)) values for p-nitrophenyl-alpha-d-galactopyranoside (pNPGal) and raffinose were 13.86 and 4.75 kcal mol(-1), respectively. The K(m) values for pNPGal, melibiose, raffinose, and stachyose were 0.45, 5.37, 39.62 and 48.80 mM, respectively. The enzyme was sensitive to inhibition by HgCl(2), SDS, AgNO(3), CuSO(4), and melibiose. d-Galactose was a competitive inhibitor (K(i)=2.74 mM). In addition to its ability to hydrolyze raffinose and stachyose, the enzyme also hydrolyzed galactomannan.
Keywords:Tachigali multijuga Benth    Fabaceae  Seed germination  α-Galactosidase  Galactomannans  Raffinose family oligosaccharides
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