Purification and properties of an extracellular conidial trehalase from Humicola grisea var. thermoidea
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A L Zimmermann1, H F Terenzi, J A Jorge
1Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Brasil.
Abstract:
An extracellular trehalase (alpha, alpha-trehalose glucohydrolase, EC 3.2.1.28) was purified from conidia of Humicola grisea var. thermoidea. The purified enzyme is a glycoprotein and migrates as a single polypeptide band during polyacrylamide gel electrophoresis under non-denaturing conditions. The apparent molecular weight of the enzyme was estimated as 580,000 by gel filtration chromatography. The enzyme is separable into three polypeptide bands of 105,000, 98,000 and 84,000 daltons on SDS-PAGE. It is specific for trehalose and its activity is not inhibited by other disaccharides. It has a Km of 2.3 mM, an optimum pH of 5.6 in sodium acetate buffer and a temperature optimum of 60 degrees C. The enzyme is activated by Ca2+, Co2+ and Mn2+ and inhibited by inorganic phosphate, AMP, ADP or ATP. The inhibitory effect of phosphate, AMP and ADP, but not that of ATP, was abolished in the presence of Ca2+.