Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland
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...
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2020
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Online Access: | https://forskning.ruc.dk/da/publications/be86750f-b806-4713-be32-b940aa564ea6 https://doi.org/10.1002/mbo3.980 https://hdl.handle.net/1800/be86750f-b806-4713-be32-b940aa564ea6 https://rucforsk.ruc.dk/ws/files/77494107/mbo3.980.pdf |
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fturoskildefispu:oai:pure.atira.dk:publications/be86750f-b806-4713-be32-b940aa564ea6 2023-05-15T16:03:40+02:00 Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland Thøgersen, Mariane S. Christensen, Stefan J. Jepsen, Morten Pedersen, Lars H. Stougaard, Peter 2020-03-01 application/pdf https://forskning.ruc.dk/da/publications/be86750f-b806-4713-be32-b940aa564ea6 https://doi.org/10.1002/mbo3.980 https://hdl.handle.net/1800/be86750f-b806-4713-be32-b940aa564ea6 https://rucforsk.ruc.dk/ws/files/77494107/mbo3.980.pdf eng eng info:eu-repo/semantics/openAccess Thøgersen , M S , Christensen , S J , Jepsen , M , Pedersen , L H & Stougaard , P 2020 , ' Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland ' , MicrobiologyOpen , vol. 9 , no. 3 , e980 . https://doi.org/10.1002/mbo3.980 galactooligosaccharides Paenibacillus transglycosylation α-l-fucosidase β-d-galactosidase article 2020 fturoskildefispu https://doi.org/10.1002/mbo3.980 2022-12-11T07:01:26Z 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 β-d-galactosidase and α-l-fucosidase activities on chromogenic substrates. This identified two genes encoding a β-d-galactosidase and an α-l-fucosidase, respectively. The enzymes were recombinantly expressed, purified, and characterized using oNPG (2-nitrophenyl-β-d-galactopyranoside) and pNP-fucose (4-nitrophenyl-α-l-fucopyranoside), respectively. The enzymes were shown to have optimal activity at 50°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 β-d-galactosidase was shown to produce the galactooligosaccharides (GOS) 6'-O-galactosyllactose and 3'-O-galactosyllactose using lactose as substrate, whereas the α-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. Article in Journal/Newspaper East Greenland Greenland Roskilde University Research Portal (RUC) Greenland MicrobiologyOpen 9 3 |
institution |
Open Polar |
collection |
Roskilde University Research Portal (RUC) |
op_collection_id |
fturoskildefispu |
language |
English |
topic |
galactooligosaccharides Paenibacillus transglycosylation α-l-fucosidase β-d-galactosidase |
spellingShingle |
galactooligosaccharides Paenibacillus transglycosylation α-l-fucosidase β-d-galactosidase Thøgersen, Mariane S. Christensen, Stefan J. Jepsen, Morten Pedersen, Lars H. Stougaard, Peter Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland |
topic_facet |
galactooligosaccharides Paenibacillus transglycosylation α-l-fucosidase β-d-galactosidase |
description |
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 β-d-galactosidase and α-l-fucosidase activities on chromogenic substrates. This identified two genes encoding a β-d-galactosidase and an α-l-fucosidase, respectively. The enzymes were recombinantly expressed, purified, and characterized using oNPG (2-nitrophenyl-β-d-galactopyranoside) and pNP-fucose (4-nitrophenyl-α-l-fucopyranoside), respectively. The enzymes were shown to have optimal activity at 50°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 β-d-galactosidase was shown to produce the galactooligosaccharides (GOS) 6'-O-galactosyllactose and 3'-O-galactosyllactose using lactose as substrate, whereas the α-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. |
format |
Article in Journal/Newspaper |
author |
Thøgersen, Mariane S. Christensen, Stefan J. Jepsen, Morten Pedersen, Lars H. Stougaard, Peter |
author_facet |
Thøgersen, Mariane S. Christensen, Stefan J. Jepsen, Morten Pedersen, Lars H. Stougaard, Peter |
author_sort |
Thøgersen, Mariane S. |
title |
Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland |
title_short |
Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland |
title_full |
Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland |
title_fullStr |
Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland |
title_full_unstemmed |
Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland |
title_sort |
transglycosylating β-d-galactosidase and α-l-fucosidase from paenibacillus sp. 3179 from a hot spring in east greenland |
publishDate |
2020 |
url |
https://forskning.ruc.dk/da/publications/be86750f-b806-4713-be32-b940aa564ea6 https://doi.org/10.1002/mbo3.980 https://hdl.handle.net/1800/be86750f-b806-4713-be32-b940aa564ea6 https://rucforsk.ruc.dk/ws/files/77494107/mbo3.980.pdf |
geographic |
Greenland |
geographic_facet |
Greenland |
genre |
East Greenland Greenland |
genre_facet |
East Greenland Greenland |
op_source |
Thøgersen , M S , Christensen , S J , Jepsen , M , Pedersen , L H & Stougaard , P 2020 , ' Transglycosylating β-d-galactosidase and α-l-fucosidase from Paenibacillus sp. 3179 from a hot spring in East Greenland ' , MicrobiologyOpen , vol. 9 , no. 3 , e980 . https://doi.org/10.1002/mbo3.980 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1002/mbo3.980 |
container_title |
MicrobiologyOpen |
container_volume |
9 |
container_issue |
3 |
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1766399370412949504 |