Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel Glycoconjugates
A few α-glucan debranching enzymes (DBEs) of the large glycoside hydrolase family 13 (GH13), also known as the α-amylase family, have been shown to catalyze transglycosylation as well as hydrolysis. However, little is known about their acceptor and donor preferences. Here, a DBE from barley, limit d...
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| Format: | Article |
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MDPI AG
2023-05-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/10/4111 |
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| author | Malene Bech Vester-Christensen Jesper Holck Martin Rejzek Léa Perrin Morten Tovborg Birte Svensson Robert A. Field Marie Sofie Møller |
| author_facet | Malene Bech Vester-Christensen Jesper Holck Martin Rejzek Léa Perrin Morten Tovborg Birte Svensson Robert A. Field Marie Sofie Møller |
| author_sort | Malene Bech Vester-Christensen |
| collection | DOAJ |
| description | A few α-glucan debranching enzymes (DBEs) of the large glycoside hydrolase family 13 (GH13), also known as the α-amylase family, have been shown to catalyze transglycosylation as well as hydrolysis. However, little is known about their acceptor and donor preferences. Here, a DBE from barley, limit dextrinase (<i>Hv</i>LD), is used as a case study. Its transglycosylation activity is studied using two approaches; (i) natural substrates as donors and different <i>p</i>-nitrophenyl (pNP) sugars as well as different small glycosides as acceptors, and (ii) α-maltosyl and α-maltotriosyl fluorides as donors with linear maltooligosaccharides, cyclodextrins, and GH inhibitors as acceptors. <i>Hv</i>LD showed a clear preference for pNP maltoside both as acceptor/donor and acceptor with the natural substrate pullulan or a pullulan fragment as donor. Maltose was the best acceptor with α-maltosyl fluoride as donor. The findings highlight the importance of the subsite +2 of <i>Hv</i>LD for activity and selectivity when maltooligosaccharides function as acceptors. However, remarkably, <i>Hv</i>LD is not very selective when it comes to aglycone moiety; different aromatic ring-containing molecules besides pNP could function as acceptors. The transglycosylation activity of <i>Hv</i>LD can provide glycoconjugate compounds with novel glycosylation patterns from natural donors such as pullulan, although the reaction would benefit from optimization. |
| first_indexed | 2024-03-11T03:27:22Z |
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| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-11T03:27:22Z |
| publishDate | 2023-05-01 |
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| record_format | Article |
| series | Molecules |
| spelling | doaj.art-620f05c062cd4838b1d09b34822420f32023-11-18T02:39:25ZengMDPI AGMolecules1420-30492023-05-012810411110.3390/molecules28104111Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel GlycoconjugatesMalene Bech Vester-Christensen0Jesper Holck1Martin Rejzek2Léa Perrin3Morten Tovborg4Birte Svensson5Robert A. Field6Marie Sofie Møller7Enzyme and Protein Chemistry, Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, DenmarkEnzyme Technology, Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, DenmarkDepartment of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7TJ, UKApplied Molecular Enzyme Chemistry, Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, DenmarkNovozymes A/S, DK-2800 Kongens Lyngby, DenmarkEnzyme and Protein Chemistry, Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, DenmarkDepartment of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7TJ, UKApplied Molecular Enzyme Chemistry, Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, DenmarkA few α-glucan debranching enzymes (DBEs) of the large glycoside hydrolase family 13 (GH13), also known as the α-amylase family, have been shown to catalyze transglycosylation as well as hydrolysis. However, little is known about their acceptor and donor preferences. Here, a DBE from barley, limit dextrinase (<i>Hv</i>LD), is used as a case study. Its transglycosylation activity is studied using two approaches; (i) natural substrates as donors and different <i>p</i>-nitrophenyl (pNP) sugars as well as different small glycosides as acceptors, and (ii) α-maltosyl and α-maltotriosyl fluorides as donors with linear maltooligosaccharides, cyclodextrins, and GH inhibitors as acceptors. <i>Hv</i>LD showed a clear preference for pNP maltoside both as acceptor/donor and acceptor with the natural substrate pullulan or a pullulan fragment as donor. Maltose was the best acceptor with α-maltosyl fluoride as donor. The findings highlight the importance of the subsite +2 of <i>Hv</i>LD for activity and selectivity when maltooligosaccharides function as acceptors. However, remarkably, <i>Hv</i>LD is not very selective when it comes to aglycone moiety; different aromatic ring-containing molecules besides pNP could function as acceptors. The transglycosylation activity of <i>Hv</i>LD can provide glycoconjugate compounds with novel glycosylation patterns from natural donors such as pullulan, although the reaction would benefit from optimization.https://www.mdpi.com/1420-3049/28/10/4111glycoside hydrolase family 13α-glucan debranching enzymeCAZyme profilingpullulancyclodextrinsglycosyl fluorides |
| spellingShingle | Malene Bech Vester-Christensen Jesper Holck Martin Rejzek Léa Perrin Morten Tovborg Birte Svensson Robert A. Field Marie Sofie Møller Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel Glycoconjugates Molecules glycoside hydrolase family 13 α-glucan debranching enzyme CAZyme profiling pullulan cyclodextrins glycosyl fluorides |
| title | Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel Glycoconjugates |
| title_full | Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel Glycoconjugates |
| title_fullStr | Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel Glycoconjugates |
| title_full_unstemmed | Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel Glycoconjugates |
| title_short | Exploration of the Transglycosylation Activity of Barley Limit Dextrinase for Production of Novel Glycoconjugates |
| title_sort | exploration of the transglycosylation activity of barley limit dextrinase for production of novel glycoconjugates |
| topic | glycoside hydrolase family 13 α-glucan debranching enzyme CAZyme profiling pullulan cyclodextrins glycosyl fluorides |
| url | https://www.mdpi.com/1420-3049/28/10/4111 |
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