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The present study was conducted to screen for psychrophilic micro-organisms that are able to hydrolyse lactose at low temperature, and to examine the cold-active β-galactosidase produced by the isolated psychrophilic micro-organisms.Psychrophilic bacteria, which grow on lactose as a sole carbon source, were isolated from soil from Hokkaido, Japan. The phenotype and sequence of 16S rDNA of the isolated strains indicated a taxonomic affiliation to Arthrobacter psychrolactophilus. The isolated A. psychrolactophilus strains were able to grow on lactose at below 5°C, and showed cold-active β-galactosidase activity, which was highly specific at even 0°C.Facts in this study may indicate the possibility that the isolated strains produce novel β-galactosidases that are able to hydrolyse lactose at low temperature, although some strains have isozymes.It may be possible that the cold active β-galactosidases from the isolated strains can be applied to the food industry, e.g. processing of milk and whey below 5°C.