Characterization of a fungal α-galactosidase and its synergistic effect with ß-mannanase for hydrolysis of galactomannan.

An acid stable α-galactosidase was produced and purified from mannolytic fungal strain, Penicillium aculeatum APS1. Enzyme was produced using wheat bran and copra cake moistened with corn steep liquor by solid state fermentation. APS1αgal having molecular weight of 65.4 kDa was purified to electrophoretic homogeneity by three phase partitioning and gel permeation chromatography with high enzyme recovery. APS1αgal was found to be maximally active at 55 °C and pH 4.5, having high stability at acidic pH. Thermal stability and thermal inactivation kinetics of APS1αgal were also studied. APS1αgal was found to effectively hydrolyse oligosaccharides as well as polysaccharides having α-1,6 linked galactose. Abolishment of enzyme activity in N-brommosuccinimide revealed an important role of tryptophan residue in catalysis. APS1αgal had shown outstanding tolerance to NaCl and proteases. MALDI-TOF MS/MS analysis indicated that enzyme is probably a member of family GH27. Synergistic interaction between APS1αgal and ß-mannanase for hydrolysis of galactomannan was very clear and maximum 2.0° of synergy was found under simultaneous mode of action. This study reports a new source of α-galactosidase with biochemical properties suitable for applications in food and feed industries.

Thermotolerant and protease-resistant GH5 family ß-mannanase with CBM1 from Penicillium aculeatum APS1: purification and characterization.

In past several years, mannanases has attracted many researchers owing to its extensive industrial applications. The search for novel mannanases with high stability still continues. Present investigation was focused on purification of extracellular ß-mannanase from Penicillium aculeatum APS1 and its characterization. APS1 mannanase was purified to homogeneity by chromatography techniques. Protein identification by MALDI-TOF MS/MS revealed that the enzyme belongs to GH family 5 and subfamily 7, and possesses CBM1. The molecular weight was found to be 40.6 kDa. The optimum temperature and pH of APS1 mannanase were 70 °C and 5.5, respectively. APS1 mannanase was found to be highly stable at 50 °C and tolerant at 55-60 °C. The enzyme was very sensitive to Mn+2, Hg+2 and Co+2 metal ions and stimulated by Zn+2. Inhibition of activity by N-bromosuccinimide suggested key role of tryptophan residues for catalytic activity. The purified enzyme was efficient in hydrolysis of locust bean gum, guar gum and konjac gum and kinetic studies revealed highest affinity towards locust bean gum (LBG). APS1 mannanase was found to be protease resistant. Looking at the properties, APS1 mannanase can be a valuable candidate for applications in bioconversion of mannan-rich substrates into value-added products and also in food and feed processing.