EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction

EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and... Biomass hydrolysis constitutes a bottleneck for the biotransformation of lignocellulosic residues into bioethanol and high-value products. The efficient deconstruction of polysaccharides to fermentable sugars requires multiple enzymes acting concertedly. GH43 β-xylosidases are among the most interesting enzymes involved in hemicellulose deconstruction into xylose. In this work, the structural and functional properties of β-xylosidase EcXyl43 from Enterobacter sp. were thoroughly characterized. Molecular modeling suggested a 3D structure formed by a conserved N-terminal catalytic domain linked to an ancillary C-terminal domain. Both domains resulted essential for enzymatic activity, and the role of critical residues, from the catalytic and the ancillary modules, was confirmed by mutagenesis. EcXyl43 presented β-xylosidase activity towards natural and artificial substrates while arabinofuranosidase activity was only detected on nitrophenyl α-L-arabinofuranoside (pNPA). It hydrolyzed xylobiose and purified xylooligosaccharides (XOS), up to degree of polymerization 6, with higher activity towards longer XOS. Low levels of activity on commercial xylan were also observed, mainly on the soluble fraction. The addition of EcXyl43 to GH10 and GH11 endoxylanases increased the release of xylose from xylan and pre-treated wheat straw. Additionally, EcXyl43 exhibited high efficiency and thermal stability under its optimal conditions (40 °C, pH 6.5), with a half-life of 58 h. Therefore, this enzyme could be a suitable additive for hemicellulases in long-term hydrolysis reactions. Because of its moderate inhibition by monomeric sugars but its high inhibition by ethanol, EcXyl43 could be particularly more useful in separate hydrolysis and fermentation (SHF) than in simultaneous saccharification and co-fermentation (SSCF) or consolidated bioprocessing (CBP). http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Microbiology and Biotechnology Springer Journals

EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction

Download PDF
13 pages
Read Article

Loading next page...
 
/lp/springer_journal/ecxyl43-xylosidase-molecular-modeling-activity-on-natural-and-H5aoU5SHW2
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
DOI
10.1007/s00253-018-9138-7
Publisher site
See Article on Publisher Site

Abstract

Biomass hydrolysis constitutes a bottleneck for the biotransformation of lignocellulosic residues into bioethanol and high-value products. The efficient deconstruction of polysaccharides to fermentable sugars requires multiple enzymes acting concertedly. GH43 β-xylosidases are among the most interesting enzymes involved in hemicellulose deconstruction into xylose. In this work, the structural and functional properties of β-xylosidase EcXyl43 from Enterobacter sp. were thoroughly characterized. Molecular modeling suggested a 3D structure formed by a conserved N-terminal catalytic domain linked to an ancillary C-terminal domain. Both domains resulted essential for enzymatic activity, and the role of critical residues, from the catalytic and the ancillary modules, was confirmed by mutagenesis. EcXyl43 presented β-xylosidase activity towards natural and artificial substrates while arabinofuranosidase activity was only detected on nitrophenyl α-L-arabinofuranoside (pNPA). It hydrolyzed xylobiose and purified xylooligosaccharides (XOS), up to degree of polymerization 6, with higher activity towards longer XOS. Low levels of activity on commercial xylan were also observed, mainly on the soluble fraction. The addition of EcXyl43 to GH10 and GH11 endoxylanases increased the release of xylose from xylan and pre-treated wheat straw. Additionally, EcXyl43 exhibited high efficiency and thermal stability under its optimal conditions (40 °C, pH 6.5), with a half-life of 58 h. Therefore, this enzyme could be a suitable additive for hemicellulases in long-term hydrolysis reactions. Because of its moderate inhibition by monomeric sugars but its high inhibition by ethanol, EcXyl43 could be particularly more useful in separate hydrolysis and fermentation (SHF) than in simultaneous saccharification and co-fermentation (SSCF) or consolidated bioprocessing (CBP).

Journal

Applied Microbiology and Biotechnology – Springer Journals

Published: Jun 6, 2018

References

  • Structure of the two-subsite β-D-xylosidase from Selenomonas ruminantium in complex with 1,3 bis[tris(hydroxymethyl)methylamino]propane
    Brunzelle, JS; Jordan, DB; McCaslin, DR; Olczak, A; Wawrzak, Z
  • The structure of an inverting GH43 β-xylosidase from Geobacillus stearothermophilus with its substrate reveals the role of the three catalytic residues
    Brüx, C; Ben-David, A; Shallom-Shezifi, D; Leon, M; Niefind, K; Shoham, G; Shoham, Y; Schomburg, D
  • Purification and characterization of a GH43 β-xylosidase from Enterobacter sp. identified and cloned from forest soil bacteria
    Campos, E; Negro Alvarez, M; Lorenzo, G; Gonzalez, S; Rorig, M; Talia, P; Grasso, D; Sáez, F; Manzanares Secades, P; Ballesteros Perdices, M; Cataldi, A
  • Development of a chimeric hemicellulase to enhance the xylose production and thermotolerance
    Diogo, JA; Hoffmam, ZB; Zanphorlin, LM; Cota, J; Machado, CB; Wolf, LD; Squina, F; Dámasio, A; Murakami, M; Ruller, R
  • Enzymatic deconstruction of xylan for biofuel production
    Dodd, D; Cann, IK
  • Characterization of a family 43 β-xylosidase from the xylooligosaccharide utilizing putative probiotic Weissella sp. strain 92
    Falck, P; Linares-Pastén, JA; Adlercreutz, P; Karlsson, EN
  • Paenibacillus sp. A59 GH10 and GH11 extracellular Endoxylanases: application in biomass bioconversion
    Ghio, S; Ontañon, OM; Piccinni, FE; Marrero, R; Talia, P; Grasso, DH; Campos, E
  • Hemicelluloses for fuel ethanol: a review
    Gírio, FM; Fonseca, C; Carvalheiro, F; Duarte, LC; Marques, S; Bogel-Łukasik, R
  • Variation in relative substrate specificity of bifunctional β-d-xylosidase/α-l-arabinofuranosidase by single-site mutations: roles of substrate distortion and recognition
    Jordan, DB; Li, XL
  • Highly active β-xylosidases of glycoside hydrolase family 43 operating on natural and artificial substrates
    Jordan, DB; Wagschal, K; Grigorescu, AA; Braker, JD
  • Enzyme recycling in lignocellulosic biorefineries
    Jørgensen, H; Pinelo, M
  • Characterization of two β-xylosidases from Bifidobacterium adolescentis and their contribution to the hydrolysis of prebiotic xylooligosaccharides
    Lagaert, S; Pollet, A; Delcour, JA; Lavigne, R; Courtin, CM; Volckaert, G
  • β-Xylosidases and α-L-arabinofuranosidases: accessory enzymes for arabinoxylan degradation
    Lagaert, S; Pollet, A; Courtin, CM; Volckaert, G
  • Divalent metal activation of a GH43 β-xylosidase
    Lee, CC; Braker, JD; Grigorescu, AA; Wagschal, K; Jordan, DB
  • Measurement of carboxymethylcellulase activity
    Miller, GL; Blum, R; Glennon, WE; Burton, AL
  • Functional association of catalytic and ancillary modules dictates enzymatic activity in glycoside hydrolase family 43 β-xylosidase
    Morais, S; Salama-Alber, O; Barak, Y; Hadar, Y; Wilson, DB; Lamed, R; Bayer, EA
  • The structural basis for catalysis and specificity of the Pseudomonas cellulosa alpha-glucuronidase GlcA67A
    Nurizzo, D; Nagy, T; Gilbert, HJ; Davies, GJ
  • UCSF chimera-a visualization system for exploratory research and analysis
    Pettersen, EF; Goddard, TD; Huang, CC; Couch, GS; Greenblatt, DM; Meng, EC; Ferrin, TE
  • Tailored catalysts for plant cell-wall degradation: redesigning the exo/endo preference of Cellvibrio japonicus arabinanase 43A
    Proctor, MR; Taylor EJ Nurizzo, D; Turkenburg, JP; Lloyd, RM; Vardakou, M; Davies, GJ; Gilbert, HJ
  • I-TASSER: a unified platform for automated protein structure and function prediction
    Roy, A; Kucukural, A; Zhang, Y
  • Purification and properties of an extracellular β-xylosidase from a newly isolated Fusarium proliferatum
    Saha, BC
  • Synergistic enhancement of cellobiohydrolase performance on pretreated corn Stover by addition of xylanase and esterase activities
    Selig, MJ; Knoshaug, EP; Adney, WS; Himmel, ME; Decker, SR
  • Biochemical characterization and identification of the catalytic residues of a family 43 β-D-xylosidase from Geobacillus stearothermophilus T-6
    Shallom, D; Leon, M; Bravman, T; Ben-David, A; Zaide, G; Belakhov, V; Shoham, G; Schomburg, D; Baasov, T; Shoham, Y
  • Directed evolution of GH43 β-xylosidase XylBH43 thermal stability and L186 saturation mutagenesis
    Singh, SK; Heng, C; Braker, JD; Chan, VJ; Lee, CC; Jordan, DB; Yuan, L; Wagschal, K
  • Expression in Escherichia coli and characterization of β-xylosidases GH39 and GH-43 from Bacillus halodurans C-125
    Smaali, I; Rémond, C; O’Donohue, MJ
  • MEGA6: molecular evolutionary genetics analysis version 6.0
    Tamura, K; Stecher, G; Peterson, D; Filipski, A; Kumar, S
  • Cloning of a novel gene encoding β-1,3-xylosidase from a marine bacterium, Vibrio sp. strain XY-214, and characterization of the gene product
    Umemoto, Y; Onishi, R; Araki, T
  • Biochemical and kinetic characterisation of a novel xylooligosaccharide-upregulated GH43 β-D-xylosidase/α-L-arabinofuranosidase (BXA43) from the probiotic Bifidobacterium animalis subsp. lactis BB-12
    Viborg, AH; Sørensen, KI; Gilad, O; Steen-Jensen, DB; Dilokpimol, A; Jacobsen, S; Svensson, B
  • Biochemical characterization of a novel dual-function arabinofuranosidase/xylosidase isolated from a compost starter mixture
    Wagschal, K; Heng, C; Lee, CC; Wong, DW
  • Catalytic properties of β-D-xylosidase XylBH43 from Bacillus halodurans C-125 and mutant XylBH43-W147G
    Wagschal, K; Jordan, B; Braker, JD
  • Two xylose-tolerant GH43 bifunctional β-xylosidase/α-arabinosidases and one GH11 xylanase from Humicola insolens and their synergy in the degradation of xylan
    Yang, X; Shi, P; Huang, H; Luo, H; Wang, Y; Zhang, W; Yao, B
  • The I-TASSER suite: protein structure and function prediction
    Yang, J; Yan, R; Roy, A; Xu, D; Poisson, J; Zhang, Y
  • Thermostable enzymes as biocatalysts in the biofuel industry
    Yeoman, CJ; Han, Y; Dodd, D; Schroeder, CM; Mackie, RI; Cann, IK

You’re reading a free preview. Subscribe to read the entire article.

Try 2 weeks free now

DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

or browse the journals available

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off

Sign up
for free
Start 14 day
Free Trial