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Hokkaido University Collection of Scholarly and Academic Papers >
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Characterization of an Unknown Region Linked to the Glycoside Hydrolase Family 17 beta-1,3-Glucanase of Vibrio vulnificus Reveals a Novel Glucan-Binding Domain
Title: | Characterization of an Unknown Region Linked to the Glycoside Hydrolase Family 17 beta-1,3-Glucanase of Vibrio vulnificus Reveals a Novel Glucan-Binding Domain |
Authors: | Kumagai, Yuya Browse this author →KAKEN DB | Kishimura, Hideki Browse this author →KAKEN DB | Lang, Weeranuch Browse this author | Tagami, Takayoshi Browse this author | Okuyama, Masayuki Browse this author →KAKEN DB | Kimura, Atsuo Browse this author →KAKEN DB |
Keywords: | glucanase | Vibrio | carbohydrate-binding domain | glycoside hydrolase family 17 |
Issue Date: | Apr-2022 |
Publisher: | MDPI |
Journal Title: | Marine drugs |
Volume: | 20 |
Issue: | 4 |
Start Page: | 250 |
Publisher DOI: | 10.3390/md20040250 |
Abstract: | The glycoside hydrolase family 17 beta-1,3-glucanase of Vibrio vulnificus (VvGH17) has two unknown regions in the N- and C-termini. Here, we characterized these domains by preparing mutant enzymes. VvGH17 demonstrated hydrolytic activity of beta-(1 -> 3)-glucan, mainly producing laminaribiose, but not of beta-(1 -> 3)/beta-(1 -> 4)-glucan. The C-terminal-truncated mutants (Delta C466 and Delta C441) showed decreased activity, approximately one-third of that of the WT, and Delta C415 lost almost all activity. An analysis using affinity gel containing laminarin or barley beta-glucan revealed a shift in the mobility of the Delta C466, Delta C441, and Delta C415 mutants compared to the WT. Tryptophan residues showed a strong affinity for carbohydrates. Three of four point-mutations of the tryptophan in the C-terminus (W472A, W499A, and W542A) showed a reduction in binding ability to laminarin and barley beta-glucan. The C-terminus was predicted to have a beta-sandwich structure, and three tryptophan residues (Trp472, Trp499, and Trp542) constituted a putative substrate-binding cave. Linker and substrate-binding functions were assigned to the C-terminus. The N-terminal-truncated mutants also showed decreased activity. The WT formed a trimer, while the N-terminal truncations formed monomers, indicating that the N-terminus contributed to the multimeric form of VvGH17. The results of this study are useful for understanding the structure and the function of GH17 beta-1,3-glucanases. |
Type: | article |
URI: | http://hdl.handle.net/2115/86762 |
Appears in Collections: | 水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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