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A novel cold-active β-D-galactosidase with transglycosylation activity from the Antarctic Arthrobacter sp. 32cB - gene cloning, purification and characterization
Abstract
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 257 kDa. 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-nitophenyl-β-D-galactopyranoside at that temperature, with Km values of 1.52 and 16.56 mM, and kcat values 30.55 and 31.84 s−1, respectively. Two units of the enzyme hydrolyzed 90% of the lactose in 1 mL of milk at 10 °C in 24 h. 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.
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- DOI:
- Digital Object Identifier (open in new tab) 10.1016/j.procbio.2014.09.018
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- Category:
- Articles
- Type:
- artykuł w czasopiśmie wyróżnionym w JCR
- Published in:
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PROCESS BIOCHEMISTRY
no. 49,
edition 12,
pages 2122 - 2133,
ISSN: 1359-5113 - Language:
- English
- Publication year:
- 2014
- Bibliographic description:
- Pawlak-Szukalska A., Wanarska M., Popinigis A., Kur J.: A novel cold-active β-D-galactosidase with transglycosylation activity from the Antarctic Arthrobacter sp. 32cB - gene cloning, purification and characterization// PROCESS BIOCHEMISTRY. -Vol. 49, iss. 12 (2014), s.2122-2133
- DOI:
- Digital Object Identifier (open in new tab) 10.1016/j.procbio.2014.09.018
- Verified by:
- Gdańsk University of Technology
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