Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine Residue
Starch-like polymers can be created through the use of enzymatic modification with glycogen branching enzymes (GBEs). GBEs are categorized in the glycoside hydrolase (GH) family 13 and 57. Both GH13 and GH57 GBEs exhibit branching and hydrolytic activity. While GH13 GBEs are also capable of α-1,4-tr...
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MDPI AG
2023-06-01
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| Series: | Polymers |
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| Online Access: | https://www.mdpi.com/2073-4360/15/13/2777 |
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| author | Hilda Hubertha Maria Bax Marc Jos Elise Cornelis van der Maarel Edita Jurak |
| author_facet | Hilda Hubertha Maria Bax Marc Jos Elise Cornelis van der Maarel Edita Jurak |
| author_sort | Hilda Hubertha Maria Bax |
| collection | DOAJ |
| description | Starch-like polymers can be created through the use of enzymatic modification with glycogen branching enzymes (GBEs). GBEs are categorized in the glycoside hydrolase (GH) family 13 and 57. Both GH13 and GH57 GBEs exhibit branching and hydrolytic activity. While GH13 GBEs are also capable of α-1,4-transglycosylation, it is yet unknown whether GH57 share this capability. Among the four crystal structures of GH57 GBEs that have been solved, a flexible loop with a conserved tyrosine was identified to play a role in the branching activity. However, it remains unclear whether this flexible loop is also involved in α-1,4-transglycosylation activity. We hypothesize that GH57 GBEs with the flexible loop and tyrosine are also capable of α-1,4-transglycosylation, similar to GH13 GBEs. The aim of the present study was to characterize the activity of GH57 GBEs to investigate a possible α-1,4-transglycosylation activity. Three GH57 GBEs were selected, one from <i>Thermococcus kodakarensis</i> with the flexible loop and two beta-strands; one from <i>Thermotoga maritima</i>, missing the flexible loop and beta-strands; and one from <i>Meiothermus</i> sp., missing the flexible loop but with the two beta-strands. The analysis of chain length distribution over time of modified maltooctadecaose, revealed, for the first time, that all three GH57 GBEs can generate chains longer than the substrate itself, showing that α-1,4-transglycosylation activity is generally present in GH57 GBEs. |
| first_indexed | 2024-03-11T01:31:39Z |
| format | Article |
| id | doaj.art-bfe1b936080a40de8f4ee1cfae15e524 |
| institution | Directory Open Access Journal |
| issn | 2073-4360 |
| language | English |
| last_indexed | 2024-03-11T01:31:39Z |
| publishDate | 2023-06-01 |
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| series | Polymers |
| spelling | doaj.art-bfe1b936080a40de8f4ee1cfae15e5242023-11-18T17:19:56ZengMDPI AGPolymers2073-43602023-06-011513277710.3390/polym15132777Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine ResidueHilda Hubertha Maria Bax0Marc Jos Elise Cornelis van der Maarel1Edita Jurak2Bioproduct Engineering, Engineering and Technology Institute Groningen, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The NetherlandsBioproduct Engineering, Engineering and Technology Institute Groningen, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The NetherlandsBioproduct Engineering, Engineering and Technology Institute Groningen, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The NetherlandsStarch-like polymers can be created through the use of enzymatic modification with glycogen branching enzymes (GBEs). GBEs are categorized in the glycoside hydrolase (GH) family 13 and 57. Both GH13 and GH57 GBEs exhibit branching and hydrolytic activity. While GH13 GBEs are also capable of α-1,4-transglycosylation, it is yet unknown whether GH57 share this capability. Among the four crystal structures of GH57 GBEs that have been solved, a flexible loop with a conserved tyrosine was identified to play a role in the branching activity. However, it remains unclear whether this flexible loop is also involved in α-1,4-transglycosylation activity. We hypothesize that GH57 GBEs with the flexible loop and tyrosine are also capable of α-1,4-transglycosylation, similar to GH13 GBEs. The aim of the present study was to characterize the activity of GH57 GBEs to investigate a possible α-1,4-transglycosylation activity. Three GH57 GBEs were selected, one from <i>Thermococcus kodakarensis</i> with the flexible loop and two beta-strands; one from <i>Thermotoga maritima</i>, missing the flexible loop and beta-strands; and one from <i>Meiothermus</i> sp., missing the flexible loop but with the two beta-strands. The analysis of chain length distribution over time of modified maltooctadecaose, revealed, for the first time, that all three GH57 GBEs can generate chains longer than the substrate itself, showing that α-1,4-transglycosylation activity is generally present in GH57 GBEs.https://www.mdpi.com/2073-4360/15/13/2777glycogen branching enzymealpha-1,4-transglycosylationflexible loop |
| spellingShingle | Hilda Hubertha Maria Bax Marc Jos Elise Cornelis van der Maarel Edita Jurak Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine Residue Polymers glycogen branching enzyme alpha-1,4-transglycosylation flexible loop |
| title | Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine Residue |
| title_full | Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine Residue |
| title_fullStr | Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine Residue |
| title_full_unstemmed | Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine Residue |
| title_short | Alpha-1,4-transglycosylation Activity of GH57 Glycogen Branching Enzymes Is Higher in the Absence of a Flexible Loop with a Conserved Tyrosine Residue |
| title_sort | alpha 1 4 transglycosylation activity of gh57 glycogen branching enzymes is higher in the absence of a flexible loop with a conserved tyrosine residue |
| topic | glycogen branching enzyme alpha-1,4-transglycosylation flexible loop |
| url | https://www.mdpi.com/2073-4360/15/13/2777 |
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