期刊
MICROBIOLOGYOPEN
卷 9, 期 3, 页码 -出版社
WILEY
DOI: 10.1002/mbo3.980
关键词
galactooligosaccharides; Paenibacillus; transglycosylation; alpha-l-fucosidase; beta-d-galactosidase
类别
资金
- Seoul National University
- Danish Council for Strategic Research [1308-00014B, 0652-20140008]
Thermal springs are excellent locations for discovery of thermostable microorganisms and enzymes. In this study, we identify a novel thermotolerant bacterial strain related to Paenibacillus dendritiformis, denoted Paenibacillus sp. 3179, which was isolated from a thermal spring in East Greenland. A functional expression library of the strain was constructed, and the library screened for beta-d-galactosidase and alpha-l-fucosidase activities on chromogenic substrates. This identified two genes encoding a beta-d-galactosidase and an alpha-l-fucosidase, respectively. The enzymes were recombinantly expressed, purified, and characterized using oNPG (2-nitrophenyl-beta-d-galactopyranoside) and pNP-fucose (4-nitrophenyl-alpha-l-fucopyranoside), respectively. The enzymes were shown to have optimal activity at 50 degrees C and pH 7-8, and they were able to hydrolyze as well as transglycosylate natural carbohydrates. The transglycosylation activities were investigated using TLC and HPLC, and the beta-d-galactosidase was shown to produce the galactooligosaccharides (GOS) 6'-O-galactosyllactose and 3'-O-galactosyllactose using lactose as substrate, whereas the alpha-l-fucosidase was able to transfer the fucose moiety from pNP-fuc to lactose, thereby forming 2'-O-fucosyllactose. Since enzymes that are able to transglycosylate carbohydrates at elevated temperature are desirable in many industrial processes, including food and dairy production, we foresee the potential use of enzymes from Paenibacillus sp. 3179 in the production of, for example, instant formula.
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