Exploring the sequence diversity in glycoside hydrolase family 13_18 reveals a novel glucosylglycerol phosphorylase

Exploring the sequence diversity in glycoside hydrolase family 13_18 reveals a novel... In the carbohydrate-active enzyme database, GH13_18 is a family of retaining glycoside phosphorylases that act on α-glucosides. In this work, we explored the functional diversity of this family by comparing distinctive sequence motifs in different branches of its phylogenetic tree. A glycoside phosphorylase from Marinobacter adhaerens HP15 that was predicted to have a novel function was expressed and characterised. The enzyme was found to catalyse the reversible phosphorolysis of 2-O-α-d-glucosylglycerol with retention of the anomeric configuration, a specificity that has never been described before. Homology modelling, docking and mutagenesis were performed to pinpoint particular acceptor site residues (Tyr194, Ala333, Gln336) involved in the binding of glycerol. The new enzyme specificity provides additional insights into bacterial metabolic routes, being the first report of a phosphorolytic route for glucosylglycerol in a glucosylglycerol-producing organism. Furthermore, glucosylglycerol phosphorylase might be an attractive biocatalyst for the production of the osmolyte glucosylglycerol, which is currently produced on industrial scale by exploiting a side activity of the closely related sucrose phosphorylase. Family GH13_18 has clearly proven to be more diverse than was initially assumed, and the analysis of specificity-determining sequence motifs has shown to be a straightforward and fruitful tool for enzyme discovery. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Microbiology and Biotechnology Springer Journals

Exploring the sequence diversity in glycoside hydrolase family 13_18 reveals a novel glucosylglycerol phosphorylase

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Publisher
Springer Journals
Copyright
Copyright © 2018 by Springer-Verlag GmbH Germany, part of Springer Nature
Subject
Life Sciences; Microbiology; Microbial Genetics and Genomics; Biotechnology
ISSN
0175-7598
eISSN
1432-0614
D.O.I.
10.1007/s00253-018-8856-1
Publisher site
See Article on Publisher Site

Abstract

In the carbohydrate-active enzyme database, GH13_18 is a family of retaining glycoside phosphorylases that act on α-glucosides. In this work, we explored the functional diversity of this family by comparing distinctive sequence motifs in different branches of its phylogenetic tree. A glycoside phosphorylase from Marinobacter adhaerens HP15 that was predicted to have a novel function was expressed and characterised. The enzyme was found to catalyse the reversible phosphorolysis of 2-O-α-d-glucosylglycerol with retention of the anomeric configuration, a specificity that has never been described before. Homology modelling, docking and mutagenesis were performed to pinpoint particular acceptor site residues (Tyr194, Ala333, Gln336) involved in the binding of glycerol. The new enzyme specificity provides additional insights into bacterial metabolic routes, being the first report of a phosphorolytic route for glucosylglycerol in a glucosylglycerol-producing organism. Furthermore, glucosylglycerol phosphorylase might be an attractive biocatalyst for the production of the osmolyte glucosylglycerol, which is currently produced on industrial scale by exploiting a side activity of the closely related sucrose phosphorylase. Family GH13_18 has clearly proven to be more diverse than was initially assumed, and the analysis of specificity-determining sequence motifs has shown to be a straightforward and fruitful tool for enzyme discovery.

Journal

Applied Microbiology and Biotechnology – Springer Journals

Published: Feb 22, 2018

References

  • Transglucosylation potential of six sucrose phosphorylases toward different classes of acceptors
    Aerts, D; Verhaeghe, TF; Roman, BI; Stevens, CV; Desmet, T; Soetaert, W
  • Trehalose phosphorylase from Euglena gracilis
    Belocopitow, E; Maréchal, LR
  • Determination of glucose in the presence of sucrose using glucose oxidase; effect of pH on absorption spectrum of oxidized o-dianisidine
    Blecher, M; Glassman, AB
  • Production of glucosyl glycerol by immobilized sucrose phosphorylase: options for enzyme fixation on a solid support and application in microscale flow format
    Bolivar, JM; Luley-Goedl, C; Leitner, E; Sawangwan, T; Nidetzky, B
  • The Carbohydrate-Active EnZymes database (CAZy): an expert resource for glycogenomics
    Cantarel, BL; Coutinho, PM; Rancurel, C; Bernard, T; Lombard, V; Henrissat, B
  • Operational stability of immobilized sucrose phosphorylase: continuous production of α-glucose-1-phosphate at elevated temperatures
    Winter, K; Cerdobbel, A; Soetaert, W; Desmet, T
  • Enzymatic glycosylation of phenolic antioxidants: phosphorylase-mediated synthesis and characterization
    Winter, K; Dewitte, G; Dirks-Hofmeister, ME; Laet, S; Pelantová, H; Křen, V; Desmet, T
  • Enzymatic glycosyl transfer: mechanisms and applications
    Desmet, T; Soetaert, W
  • Enzymatic glycosylation of small molecules: challenging substrates require tailored catalysts
    Desmet, T; Soetaert, W; Bojarová, P; Křen, V; Dijkhuizen, L; Eastwick-Field, V; Schiller, A
  • Creating space for large acceptors: rational biocatalyst design for resveratrol glycosylation in an aqueous system
    Dirks-Hofmeister, ME; Verhaeghe, T; De Winter, K; Desmet, T
  • Glucosylglycerol, a compatible solute, sustains cell division under salt stress
    Ferjani, A; Mustardy, L; Sulpice, R; Marin, K; Suzuki, I; Hagemann, M; Murata, N
  • Glucosylglycerate phosphorylase, an enzyme with novel specificity involved in compatible solute metabolism
    Franceus, J; Pinel, D; Desmet, T
  • Microtiter assay for glutamine synthetase biosynthetic activity using inorganic phosphate detection
    Gawronski, JD; Benson, DR
  • A high-yielding biocatalytic process for the production of 2-O-(alpha-D-glucopyranosyl)-sn-glycerol, a natural osmolyte and useful moisturizing ingredient
    Goedl, C; Sawangwan, T; Mueller, M; Schwarz, A; Nidetzky, B
  • New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0
    Guindon, S; Dufayard, J-F; Lefort, V; Anisimova, M; Hordijk, W; Gascuel, O
  • The plant-associated bacterium Stenotrophomonas rhizophila expresses a new enzyme for the synthesis of the compatible solute glucosylglycerol
    Hagemann, M; Ribbeck-busch, K; Kla, S; Hasse, D; Steinbruch, R; Berg, G
  • Biosynthesis of glucosyl glycerol, a compatible solute, using intermolecular transglycosylation activity of amylosucrase from Methylobacillus flagellatus KT
    Jeong, J; Seo, D; Jung, J; Park, J; Baek, N; Kim, M; Park, C
  • Marinobacter adhaerens sp. nov., isolated from marine aggregates formed with the diatom Thalassiosira weissflogii
    Kaeppel, EC; Ga, A; Seebah, S; Grossart, H; Ullrich, MS
  • Uptake and synthesis of compatible solutes as microbial stress responses to high-osmolality environments
    Kempf, B; Bremer, E
  • Diversity of phosphorylases in glycoside hydrolase families
    Kitaoka, M
  • Extremolytes: natural compounds from extremophiles for versatile applications
    Lentzen, G; Schwarz, T
  • The carbohydrate-active enzymes database (CAZy) in 2013
    Lombard, V; Golaconda Ramulu, H; Drula, E; Coutinho, PM; Henrissat, B
  • DOOR 2.0: presenting operons and their functions through dynamic and integrated views
    Mao, X; Ma, Q; Zhou, C; Chen, X; Zhang, H; Yang, J; Mao, F; Lai, W; Xu, Y
  • Dissecting differential binding of fructose and phosphate as leaving group/nucleophile of glucosyl transfer catalyzed by sucrose phosphorylase
    Mueller, M; Nidetzky, B
  • The role of Asp-295 in the catalytic mechanism of Leuconostoc mesenteroides sucrose phosphorylase probed with site-directed mutagenesis
    Mueller, M; Nidetzky, B
  • 2-O-α-D-Glucosylglycerol phosphorylase from Bacillus selenitireducens MLS10 possessing hydrolytic activity on β-D-glucose 1-phosphate
    Nihira, T; Saito, Y; Ohtsubo, K; Nakai, H; Kitaoka, M
  • Glucosylglycerate metabolism, bioversatility and mycobacterial survival
    Nunes-Costa, D; Maranha, A; Costa, M; Alarico, S; Empadinhas, N
  • Sucrose utilisation in bacteria: genetic organisation and regulation
    Reid, SJ; Abratt, VR
  • Purification and characterization of a trehalose synthase from the basidiomycete Grifola frondosa
    Saito, K; Kase, T; Takahashi, E; Takahashi, E; Horinouchi, S
  • Improved PCR method for the creation of saturation mutagenesis libraries in directed evolution: application to difficult-to-amplify templates
    Sanchis, J; Fernández, L; Carballeira, JD; Drone, J; Gumulya, Y; Höbenreich, H; Kahakeaw, D; Kille, S; Lohmer, R; Peyralans, JJ-P; Podtetenieff, J; Prasad, S; Soni, P; Taglieber, A; Wu, S; Zilly, FE; Reetz, MT
  • Glucosylglycerol and glucosylglycerate as enzyme stabilizers
    Sawangwan, T; Goedl, C; Nidetzky, B
  • Trehalose phosphorylase from Pleurotus ostreatus: characterization and stabilization by covalent modification, and application for the synthesis of α,α-trehalose
    Schwarz, A; Goedl, C; Minani, A; Nidetzky, B
  • Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega
    Sievers, F; Wilm, A; Dineen, D; Gibson, TJ; Karplus, K; Li, W; Lopez, R; McWilliam, H; Remmert, M; Söding, J; Thompson, JD; Higgins, DG
  • Purification and mechanism of action of sucrose phosphorylase
    Silverstein, R; Voet, J; Reed, D; Abeles, R
  • Dividing the large glycoside hydrolase family 13 into subfamilies: towards improved functional annotations of α-amylase-related proteins
    Stam, MR; Danchin, EGJ; Rancurel, C; Coutinho, PM; Henrissat, B
  • Synthesis of alpha-D-glucosylglycerol by alpha-glucosidase and some of its characteristics
    Takenaka, F; Uchiyama, H
  • AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading
    Trott, O; Olson, A
  • Mapping the acceptor site of sucrose phosphorylase from Bifidobacterium adolescentis by alanine scanning
    Verhaeghe, T; Diricks, M; Aerts, D; Soetaert, W; Desmet, T
  • The quest for a thermostable sucrose phosphorylase reveals sucrose 6′-phosphate phosphorylase as a novel specificity
    Verhaeghe, T; Aerts, D; Diricks, M; Soetaert, W; Desmet, T

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