Isolation and identification of a strain producing cold-adapted β galactosidase, and purification and characterisation of the enzyme
W.Y. Liu, Y.W. Shi, X.Q. Wang, K. Louhttps://doi.org/10.17221/31/2008-CJFSCitation:Liu W.Y., Shi Y.W., Wang X.Q., Lou K. (2008): Isolation and identification of a strain producing cold-adapted β galactosidase, and purification and characterisation of the enzyme. Czech J. Food Sci., 26: 284-290.
Enzymes with high specific activities at low temperatures have potential uses in the food industry. Cold-adapted microorganisms are potentially useful sources of cold-active enzyme. To find cold-adapted β-galactosidase, we isolated several cold-adapted microorganisms from glacier zone soil. One cold-adapted β-galactosidase producing strain was obtained. The biochemical characteristics and the results of 16S rDNA sequencing identified the strain as Rahnella aquatilis. The enzyme was purified by column chromatography after which a single protein band migrating near 60 kDa was observed by means of SDS-PAGE. The β-galactosidase was optimally active at 35°C and at pH 6.5 when assayed with o-nitrophenyl-β-D-galactopyrano-side as substrate. The enzyme activity was sensitive to temperatures above 40°C and was undetectable at 45°C. Metal ions Mn2+and K+ activated the enzyme while Cu2+, Zn2+, Fe3+, and Al3+ inhibited the activity. The enzyme was also assayed for lactose hydrolysis. When milk is treated with the enzyme at 30°C for 2 h, the degree of lactose hydrolysis can reach 80%. It has, thus, potential applications in the food industry.Keywords:psychrotrophic microorganism; cold-adapted β-galactosidase; lactose hydrolysis