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Heterologous Production, Purification and Characterization of Two Cold-Active β- D-Galactosidases with Transglycosylation Activity from the Psychrotolerant Arctic Bacterium Arthrobacter sp. S3* Isolated from Spitsbergen Island Soil
Abstract
Cold-adapted microorganisms possess cold-active enzymes with potential applications in different industries and research areas. In this study, two genes encoding β-D-galactosidases belonging to Glycoside Hydrolase families 2 and 42 from the psychrotolerant Arctic bacterium Arthrobacter sp. S3* were cloned, expressed in Escherichia coli and Komagataella phaffii, purified and characterized. The GH2 β-D-galactosidase is a tetramer with a molecular weight of 450 kDa, while the GH42 β-D-galactosidase is a 233 kDa trimer. The Bgal2 was optimally active at pH 7.5 and 22 °C and maintained 57% of maximum activity at 10 °C, whereas the Bgal42 was optimally active at pH 7.0 and 40 °C and exhibited 44% of maximum activity at 10 °C. Both enzymes hydrolyzed lactose and showed transglycosylation activity. We also found that 2 U/mL of the Bgal2 hydrolyzed 85% of lactose in milk within 10 h at 10 °C. The enzyme synthesized galactooligosaccharides, heterooligosaccharides, alkyl galactopyranosides and glycosylated salicin. The Bgal42 synthesized galactooligosaccharides and 20 U/mL of the enzyme hydrolyzed 72% of milk lactose within 24 h at 10 °C. The properties of Arthrobacter sp. S3* Bgal2 make it a candidate for lactose hydrolysis in the dairy industry and a promising tool for the glycosylation of various acceptors in the biomedical sector.
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- Category:
- Articles
- Type:
- artykuły w czasopismach
- Published in:
-
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
no. 25,
ISSN: 1661-6596 - Language:
- English
- Publication year:
- 2024
- Bibliographic description:
- Wanarska M., Pawlak-Szukalska A., Rosińska A., Kozłowska-Tylingo K.: Heterologous Production, Purification and Characterization of Two Cold-Active β- D-Galactosidases with Transglycosylation Activity from the Psychrotolerant Arctic Bacterium Arthrobacter sp. S3* Isolated from Spitsbergen Island Soil// INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES -,iss. 25/13354 (2024), s.1-28
- DOI:
- Digital Object Identifier (open in new tab) 10.3390/ijms252413354
- Sources of funding:
-
- Projekt nr 037646, kierownik: dr hab. inż. Robert Tylingo, prof. uczelni, prace zlecone.
- Verified by:
- Gdańsk University of Technology
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