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Updated: Jul 5, 2025
06:51
High-throughput Screening of Carbohydrate-degrading Enzymes Using Novel Insoluble Chromogenic Substrate Assay Kits
Published on: September 20, 2016
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Mapping secondary substrate-binding sites on the GH11 xylanase from Bacillus subtilis
Gustavo Avelar Molina1, Luis Felipe Santos Mendes2, Carlos Alessandro Fuzo1,3
1Department of Chemistry, Faculty of Philosophy, Sciences and Literature at Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil.
FEBS Letters
|January 22, 2024
Summary
Bacillus subtilis xylanase regulation was studied using molecular dynamics and experiments. A second allosteric binding site near Asn151 was discovered, aiding biomass conversion.
Area of Science:
- Biochemistry
- Enzymology
- Biotechnology
Background:
- Xylanases are crucial enzymes for biomass conversion.
- Understanding their regulation is key for industrial applications and sustainable energy.
- Bacillus subtilis GH11 endoxylanase is a model system for studying xylan hydrolysis.
Purpose of the Study:
- To investigate the allosteric regulation of Bacillus subtilis GH11 endoxylanase.
- To identify and characterize secondary binding sites (SBS) involved in xylanase regulation.
- To provide insights into enzyme mechanisms for improved biomass conversion technologies.
Main Methods:
- Molecular dynamics simulations (MDS) with xylobiose.
- Arabinoxylan titration experiments.
- Site-directed mutagenesis of cysteine residues (N54C, N151C).
- Labeling with acrylodan and ESR spin-label MTSSL.
Main Results:
- MDS identified active site and two potential SBS around Asn54 and Asn151.
- Experimental validation confirmed ligand binding at SBS near Asn54.
- A novel SBS near Asn151 was discovered, involving residues Val98, Ala192, Ser155, and His156.
Conclusions:
- Allosteric regulation of Bacillus subtilis xylanase involves multiple binding sites.
- The newly identified SBS near Asn151 offers new targets for enzyme modulation.
- This research contributes to industrial decarbonization and sustainable energy solutions.
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