Pawlak-Szukalska A.,Gdan sk University of Technology |
Wanarska M.,Gdan sk University of Technology |
Popinigis A.T.,Gdan sk University of Technology |
Kur J.,Gdan sk University of Technology
Process Biochemistry | Year: 2014
A gene encoding a novel β-d-galactosidase from the psychrotolerant Antarctic bacterium Arthrobacter sp. 32cB was isolated, cloned and expressed in Escherichia coli. The active form of recombinant β-d-galactosidase consists of two subunits with a combined molecular weight of approximately 257kDa. The enzyme's maximum activity towards o-nitrophenyl-β-d-galactopyranoside was determined as occurring at 28°C and pH 8.0. However, it exhibited 42% of maximum activity at 10°C and was capable of hydrolyzing both lactose and o-nitrophenyl-β-d-galactopyranoside at that temperature, with K m values of 1.52 and 16.56mM, and k cat values 30.55 and 31.84s-1, respectively. Two units of the enzyme hydrolyzed 90% of the lactose in 1mL of milk at 10°C in 24h. The transglycosylation activity of Arthrobacter sp. 32cB β-d-galactosidase was also examined. It synthesized galactooligosaccharides in a temperature range from 10 to 30°C. Moreover, it catalyzed the synthesis of heterooligosaccharides such as lactulose, galactosyl-xylose and galactosyl-arabinose, alkyl glycosides, and glycosylated salicin from lactose and the appropriate acceptor at 30°C. The properties of Arthrobacter sp. 32cB β-d-galactosidase make it a candidate for use in the industrial removal of lactose from milk and a promising tool for the glycosylation of various acceptors, especially those which are thermosensitive. © 2014 The Authors.