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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 p-galactosidase, we isolated several cold-adapted microorganisms from glacier zone soil. One cold-adapted p-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 p-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 Mn 2+ and K+ activated the enzyme while Cu 2+ , Zn 2+ , Fe 3 +, and Al 3+ 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.

Isolation and identification of a strain producing cold-adapted β galactosidase, and purification and characterisation of the enzyme