PURIFICATION AND SOME PROPERTIES OF ALPHA-GALACTOSIDASE FROM PSEUDOMONAS-FLUORESCENS H-601

Pseudomonas fluorescens H-601, isolated from soil, produces alpha-galactosidase. The enzyme was purified to homogeneity by disc electrophoresis after column chromatographies on Butyl-Toyopearl 650M, DEAE-Toyopearl 650M, and Toyopearl HW-55F. The enzyme had a molecular weight of 390,000 by gel filtration with Toyopearl HW-55F and 86,000 by SDS-polyacrylamide gel electrophoresis, and an isoelectric point of 6.3. The enzyme was most active at pH 6.0-7.0 and at 45-degrees-C and stable up to 40-degrees-C at pH 6.5 for 15 min of incubation. The enzyme hydrolyzed PNP alpha-galactoside, ONP alpha-galactoside, melibiose, raffinose, and stachyose at the relative velocities of 100, 59, 28, 13, and 12. The enzyme had strong transfer activity and wide acceptor specificity.