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Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome

Ruminant animals rely on the activities of β-glucosidases from residential microbes to convert feed fibers into glucose for further metabolic uses. In this report, we determined the structures of Br2, which is a glycoside hydrolase family 1 β-glucosidase from the bovine rumen metagenome. Br2 folds i...

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Autores principales: Kaenying, Wilaiwan, Tagami, Takayoshi, Suwan, Eukote, Pitsanuwong, Chariwat, Chomngam, Sinchai, Okuyama, Masayuki, Kongsaeree, Palangpon, Kimura, Atsuo, Kongsaeree, Prachumporn T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10685196/
https://www.ncbi.nlm.nih.gov/pubmed/38034805
http://dx.doi.org/10.1016/j.heliyon.2023.e21923
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author Kaenying, Wilaiwan
Tagami, Takayoshi
Suwan, Eukote
Pitsanuwong, Chariwat
Chomngam, Sinchai
Okuyama, Masayuki
Kongsaeree, Palangpon
Kimura, Atsuo
Kongsaeree, Prachumporn T.
author_facet Kaenying, Wilaiwan
Tagami, Takayoshi
Suwan, Eukote
Pitsanuwong, Chariwat
Chomngam, Sinchai
Okuyama, Masayuki
Kongsaeree, Palangpon
Kimura, Atsuo
Kongsaeree, Prachumporn T.
author_sort Kaenying, Wilaiwan
collection PubMed
description Ruminant animals rely on the activities of β-glucosidases from residential microbes to convert feed fibers into glucose for further metabolic uses. In this report, we determined the structures of Br2, which is a glycoside hydrolase family 1 β-glucosidase from the bovine rumen metagenome. Br2 folds into a classical (β/α)(8)-TIM barrel domain but displays unique structural features at loop β5→α5 and α-helix 5, resulting in different positive subsites from those of other GH1 enzymes. Br2 exhibited the highest specificity toward laminaritriose, suggesting its involvement in β-glucan hydrolysis in digested feed. We then substituted the residues at subsites +1 and + 2 of Br2 with those of Halothermothrix orenii β-glucosidase. The C170E and C221T mutations provided favorable interactions with glucooligosaccharide substrates at subsite +2, while the A219N mutation probably improved the substrate preference for cellobiose and gentiobiose relative to laminaribiose at subsite +1. The N407Y mutation increased the affinity toward cellooligosaccharides. These results give further insights into the molecular determinants responsible for substrate specificity in GH1 β-glucosidases and may provide a basis for future enzyme engineering applications.
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spelling pubmed-106851962023-11-30 Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome Kaenying, Wilaiwan Tagami, Takayoshi Suwan, Eukote Pitsanuwong, Chariwat Chomngam, Sinchai Okuyama, Masayuki Kongsaeree, Palangpon Kimura, Atsuo Kongsaeree, Prachumporn T. Heliyon Research Article Ruminant animals rely on the activities of β-glucosidases from residential microbes to convert feed fibers into glucose for further metabolic uses. In this report, we determined the structures of Br2, which is a glycoside hydrolase family 1 β-glucosidase from the bovine rumen metagenome. Br2 folds into a classical (β/α)(8)-TIM barrel domain but displays unique structural features at loop β5→α5 and α-helix 5, resulting in different positive subsites from those of other GH1 enzymes. Br2 exhibited the highest specificity toward laminaritriose, suggesting its involvement in β-glucan hydrolysis in digested feed. We then substituted the residues at subsites +1 and + 2 of Br2 with those of Halothermothrix orenii β-glucosidase. The C170E and C221T mutations provided favorable interactions with glucooligosaccharide substrates at subsite +2, while the A219N mutation probably improved the substrate preference for cellobiose and gentiobiose relative to laminaribiose at subsite +1. The N407Y mutation increased the affinity toward cellooligosaccharides. These results give further insights into the molecular determinants responsible for substrate specificity in GH1 β-glucosidases and may provide a basis for future enzyme engineering applications. Elsevier 2023-11-07 /pmc/articles/PMC10685196/ /pubmed/38034805 http://dx.doi.org/10.1016/j.heliyon.2023.e21923 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Kaenying, Wilaiwan
Tagami, Takayoshi
Suwan, Eukote
Pitsanuwong, Chariwat
Chomngam, Sinchai
Okuyama, Masayuki
Kongsaeree, Palangpon
Kimura, Atsuo
Kongsaeree, Prachumporn T.
Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome
title Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome
title_full Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome
title_fullStr Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome
title_full_unstemmed Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome
title_short Structural and mutational analysis of glycoside hydrolase family 1 Br2 β-glucosidase derived from bovine rumen metagenome
title_sort structural and mutational analysis of glycoside hydrolase family 1 br2 β-glucosidase derived from bovine rumen metagenome
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10685196/
https://www.ncbi.nlm.nih.gov/pubmed/38034805
http://dx.doi.org/10.1016/j.heliyon.2023.e21923
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