scholarly journals Functional Association of Catalytic and Ancillary Modules Dictates Enzymatic Activity in Glycoside Hydrolase Family 43 β-Xylosidase

2012 ◽  
Vol 287 (12) ◽  
pp. 9213-9221 ◽  
Author(s):  
Sarah Moraïs ◽  
Orly Salama-Alber ◽  
Yoav Barak ◽  
Yitzhak Hadar ◽  
David B. Wilson ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Glycoside Hydrolase ◽  
Glycoside Hydrolase Family ◽  
Functional Association ◽  
Glycoside Hydrolase Family 43 ◽  
Hydrolase Family

scholarly journals Biochemical characterization of a glycoside hydrolase family 43 β-D-galactofuranosidase from the fungus Aspergillus niger

2021 ◽  
Author(s):  
Gregory S Bulmer ◽  
Fang Wei Yuen ◽  
Naimah Begum ◽  
Bethan S Jones ◽  
Sabine S Flitsch ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Glycoside Hydrolase ◽  
Biochemical Characterization ◽  
Maximum Activity ◽  
Glycoside Hydrolase Family ◽  
Enzyme Specificity ◽  
Fungal Enzymes ◽  
Glycoside Hydrolase Family 43 ◽  
Hydrolase Family

β-D-Galactofuranose (Galf) and its polysaccharides are found in bacteria, fungi and protozoa but do not occur in mammalian tissues, and thus represent a specific target for anti-pathogenic drugs. Understanding the enzymatic degradation of these polysaccharides is therefore of great interest, but the identity of fungal enzymes with exclusively galactofuranosidase activity has so far remained elusive. Here we describe the identification and characterization of a galactofuranosidase from the industrially important fungus Aspergillus niger. Phylogenetic analysis of glycoside hydrolase family 43 subfamily 34 (GH43_34) members revealed the occurrence of three distinct clusters and, by comparison with specificities of characterized bacterial members, suggested a basis for prediction of enzyme specificity. Using this rationale, in tandem with molecular docking, we identified a putative β-D-galactofuranosidase from A. niger which was recombinantly expressed in Escherichia coli. The Galf-specific hydrolase, encoded by xynD demonstrates maximum activity at pH 5, 25 °C towards 4-Nitrophenyl-β-galactofuranoside (pNP-βGalf), with a Km of 17.9 ± 1.9 mM and Vmax of 70.6 ± 5.3 μmol min-1. The characterization of this first fungal GH43 galactofuranosidase offers further molecular insight into the degradation of Galf-containing structures and may inform clinical treatments against fungal pathogens.

A Novel Multifunctional Arabinofuranosidase/Endo-xylanase/β-Xylosidase GH43 from Paenibacillus curdlanolyticus B-6 and Its Synergistic Action to Produce Arabinose and Xylose from Cereal Arabinoxylan

2021 ◽  
Author(s):  
Puangpen Limsakul ◽  
Paripok Phitsuwan ◽  
Rattiya Waeonukul ◽  
Patthra Pason ◽  
Chakrit Tachaapaikoon ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Synergistic Action ◽  
Glycoside Hydrolase Family ◽  
Carbohydrate Binding ◽  
Glycoside Hydrolase Family 43 ◽  
Paenibacillus Curdlanolyticus ◽  
Xylanolytic Enzyme ◽  
Almost All ◽  
Xylanolytic Enzymes ◽  
Hydrolase Family

The PcAxy43B is a modular protein comprising a catalytic domain of glycoside hydrolase family 43 (GH43), a family 6 carbohydrate-binding module (CBM6) and a family 36 carbohydrate-binding module (CBM36) and found to be a novel multifunctional xylanolytic enzyme from Paenibacillus curdlanolyticus B-6. This enzyme exhibited α-L-arabinofuranosidase, endo-xylanase and β-D-xylosidase activities. α-L-Arabinofuranosidase of PcAxy43B revealed the new property of GH43, which released arabinose from the short-chain arabinoxylo-oligosaccharide (AXOS) and cereal arabinoxylan, and from both sides of the xylose residues of AXOS, which usually obstruct the action of xylanolytic enzymes. The PcAxy43B liberated series of xylo-oligosaccharides (XOSs) from birchwood xylan and xylohexaose, indicating that PcAxy43B exhibited endo-xylanase activity. The PcAxy43B produced xylose from xylobiose and reacted with p -nitrophenyl-β-D-xylopyranoside as a result of β-xylosidase activity. The PcAxy43B effectively released arabinose together with XOSs and xylose from the highly arabinosyl-substituted rye arabinoxylan. Moreover, PcAxy43B showed significant synergistic action with a trifunctional endo-xylanase/β-xylosidase/α-L-arabinofuranosidase PcAxy43A and an endo-xylanase Xyn10C from the strain B-6, in which almost all products produced from rye arabinoxylan by these combined enzymes were arabinose and xylose. In addition, the presence of CBM36 was found to be necessary for the endo-xylanase property of PcAxy43B. The PcAxy43B is capable of hydrolysing untreated cereal biomass, corn hull and rice straw into XOSs and xylose. Hence, PcAxy43B, the significant accessory multifunctional xylanolytic enzyme, is a potential candidate for application in the saccharification of cereal biomass. IMPORTANCE Enzymatic saccharification of cereal biomass is a strategy for the production of fermented sugars from low-price raw materials. In the present study, PcAxy43B from P. curdlanolyticus B-6 was found to be a novel multifunctional α-L-arabinofuranosidase/endo-xylanase/β-D-xylosidase enzyme of the glycoside hydrolase family 43. It is effective in releasing arabinose, xylose and XOSs from the highly arabinosyl-substituted rye arabinoxylan, which is usually resistant to hydrolysis by xylanolytic enzymes. Moreover, almost all products produced from rye arabinoxylan by the combination of PcAxy43B with trifunctional xylanolytic enzyme PcAxy43A and endo-xylanase Xyn10C from the strain B-6 were arabinose and xylose, which can be used to produce several value-added products. In addition, PcAxy43B is capable of hydrolysing untreated cereal biomass into XOSs and xylose. Thus, PcAxy43B is an important multifunctional xylanolytic enzyme with high potential in biotechnology.

Bacillus licheniformistrehalose-6-phosphate hydrolase structures suggest keys to substrate specificity

2016 ◽  
Vol 72 (1) ◽  
pp. 59-70 ◽  
Author(s):  
Min-Guan Lin ◽  
Meng-Chun Chi ◽  
Vankadari Naveen ◽  
Yi-Ching Li ◽  
Long-Liu Lin ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Bacillus Licheniformis ◽  
Active Site ◽  
Glycoside Hydrolase ◽  
Positive Charge ◽  
Glycoside Hydrolase Family ◽  
Erwinia Rhapontici ◽  
Hydrolysis Of ◽  
Hydrolase Family ◽  
Glycoside Hydrolase Family 13

Trehalose-6-phosphate hydrolase (TreA) belongs to glycoside hydrolase family 13 (GH13) and catalyzes the hydrolysis of trehalose 6-phosphate (T6P) to yield glucose and glucose 6-phosphate. The products of this reaction can be further metabolized by the energy-generating glycolytic pathway. Here, crystal structures ofBacillus licheniformisTreA (BlTreA) and its R201Q mutant complexed withp-nitrophenyl-α-D-glucopyranoside (R201Q–pPNG) are presented at 2.0 and 2.05 Å resolution, respectively. The overall structure ofBlTreA is similar to those of other GH13 family enzymes. However, detailed structural comparisons revealed that the catalytic site ofBlTreA contains a long loop that adopts a different conformation from those of other GH13 family members. Unlike the homologous regions ofBacillus cereusoligo-1,6-glucosidase (BcOgl) andErwinia rhaponticiisomaltulose synthase (NX-5), the surface potential of theBlTreA active site exhibits a largely positive charge contributed by the four basic residues His281, His282, Lys284 and Lys292. Mutation of these residues resulted in significant decreases in the enzymatic activity ofBlTreA. Strikingly, the281HHLK284motif and Lys292 play critical roles in substrate discrimination byBlTreA.

scholarly journals Dividing the Large Glycoside Hydrolase Family 43 into Subfamilies: a Motivation for Detailed Enzyme Characterization

2016 ◽  
Vol 82 (6) ◽  
pp. 1686-1692 ◽  
Author(s):  
Keith Mewis ◽  
Nicolas Lenfant ◽  
Vincent Lombard ◽  
Bernard Henrissat
Keyword(s):  
Glycoside Hydrolase ◽  
Enzyme Characterization ◽  
Glycoside Hydrolase Family ◽  
Structural Studies ◽  
Conserved Sequence ◽  
Functional Differences ◽  
Biochemical Assays ◽  
Glycoside Hydrolase Family 43 ◽  
Subfamily Classification ◽  
Hydrolase Family

ABSTRACTThe rapid rise in DNA sequencing has led to an expansion in the number of glycoside hydrolase (GH) families. The GH43 family currently contains α-l-arabinofuranosidase, β-d-xylosidase, α-l-arabinanase, and β-d-galactosidase enzymes for the debranching and degradation of hemicellulose and pectin polymers. Many studies have revealed finer details about members of GH43 that necessitate the division of GH43 into subfamilies, as was done previously for the GH5 and GH13 families. The work presented here is a robust subfamily classification that assigns over 91% of all complete GH43 domains into 37 subfamilies that correlate with conserved sequence residues and results of biochemical assays and structural studies. Furthermore, cooccurrence analysis of these subfamilies and other functional modules revealed strong associations between some GH43 subfamilies and CBM6 and CBM13 domains. Cooccurrence analysis also revealed the presence of proteins containing up to three GH43 domains and belonging to different subfamilies, suggesting significant functional differences for each subfamily. Overall, the subfamily analysis suggests that the GH43 enzymes probably display a hitherto underestimated variety of subtle specificity features that are not apparent when the enzymes are assayed with simple synthetic substrates, such as pNP-glycosides.

β-d-Xylosidase From Selenomonas ruminantium of Glycoside Hydrolase Family 43

2007 ◽  
pp. 93-104
Author(s):  
Douglas B. Jordan ◽  
Xin-Liang Li ◽  
Christopher A. Dunlap ◽  
Terence R. Whitehead ◽  
Michael A. Cotta
Keyword(s):  
Glycoside Hydrolase ◽  
Glycoside Hydrolase Family ◽  
Selenomonas Ruminantium ◽  
Glycoside Hydrolase Family 43 ◽  
Hydrolase Family

scholarly journals Structural analysis of a glycoside hydrolase family 43 arabinoxylan arabinofuranohydrolase in complex with xylotetraose reveals a different binding mechanism compared with other members of the same family

2009 ◽  
Vol 418 (1) ◽  
pp. 39-47 ◽  
Author(s):  
Elien Vandermarliere ◽  
Tine M. Bourgois ◽  
Martyn D. Winn ◽  
Steven van Campenhout ◽  
Guido Volckaert ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Catalytic Domain ◽  
Crystallographic Analysis ◽  
Glycoside Hydrolase Family ◽  
Carbohydrate Binding ◽  
Carbohydrate Binding Module ◽  
Stacking Interactions ◽  
Glycoside Hydrolase Family 43 ◽  
Arabinoxylan Arabinofuranohydrolase ◽  
Hydrolase Family

AXHs (arabinoxylan arabinofuranohydrolases) are α-L-arabinofuranosidases that specifically hydrolyse the glycosidic bond between arabinofuranosyl substituents and xylopyranosyl backbone residues of arabinoxylan. Bacillus subtilis was recently shown to produce an AXH that cleaves arabinose units from O-2- or O-3-mono-substituted xylose residues: BsAXH-m2,3 (B. subtilis AXH-m2,3). Crystallographic analysis reveals a two-domain structure for this enzyme: a catalytic domain displaying a five-bladed β-propeller fold characteristic of GH (glycoside hydrolase) family 43 and a CBM (carbohydrate-binding module) with a β-sandwich fold belonging to CBM family 6. Binding of substrate to BsAXH-m2,3 is largely based on hydrophobic stacking interactions, which probably allow the positional flexibility needed to hydrolyse both arabinose substituents at the O-2 or O-3 position of the xylose unit. Superposition of the BsAXH-m2,3 structure with known structures of the GH family 43 exo-acting enzymes, β-xylosidase and α-L-arabinanase, each in complex with their substrate, reveals a different orientation of the sugar backbone.

Sign in / Sign up

Export Citation Format

Share Document