Journal of Chemical and Pharmaceutical Research (ISSN : 0975-7384)

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Original Articles: 2016 Vol: 8 Issue: 7

Purification and characterization of a-Galactosidase produced from mutant bacterial strain

Abstract

An organism producing extracellular α-Galactosidase was isolated from soil and identified as Bacillus circulans. An extracellular α-Galactosidase from Bacillus circulans has been purified to homogeneity and purity by chromatographic steps, using Sephadex G-100, DEAE-cellulos, and Sephadex G-200. The specific activity of the enzyme was increased approximately 9.5 fold. The molecular weight of the purified enzyme was determined by SDSPAGE was 81 kDa. The optimum pH and temperature for the purified enzyme were 4.5 and 40ºC, respectively. α- Galactosidase is stable over abroad pH rang (3.0-10.0). Km values for p-nitrophenyl-a-D-galactopyranoside, melibiose, and raffinose were determined 2.00, 50.0, and 97.9 mM, respectively. α-Galactosidase activity was strongly inhibited by Ag+ and Hg2+ metal ions. The enzyme hydrolyzes flatulence causing galactooligosaccharides, viz. raffinose and stachyose present in the soybean