A Machine Learning Approach for the Prediction of Thermostable β-Glucosidases
Source abstract
Abstract
Thermostable β-glucosidases (E.C. 3.2.1.21) are essential enzymes used in second-generation biofuel production. However, little is known about the structural characteristics that lead to their thermostability. In this study, I used graph-based structural signatures to represent three-dimensional structures of β-glucosidase enzymes extracted from thermophilic organisms. I collected 1717 structures from thermophilic (n = 890) and non-thermophilic (n = 827) organisms and divided them into two datasets: training (n = 1134) and test (n = 583). I then used seven machine learning algorithms to classify them. The best model achieved 77.1% accuracy using logistic regression in training with 10-fold cross-validation and 81.6% accuracy in testing using the CatBoost algorithm. I hypothesize that the signature model proposed here can help understand the structural patterns in thermostable enzymes and shed light on the design of more efficient enzymes for biofuel production.
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Research context
OpenAlex topics: Microbial Metabolic Engineering and Bioproduction · Biofuel production and bioconversion · Machine Learning in Bioinformatics
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Department of Computer Science (DCC), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte 31270-901, Brazil
Funding
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES)
References (54)
Show source-provided references
1. Heterologous Expression in Pichia Pastoris and Characterization of an Endogenous Thermostable and High-Glucose-Tolerant β-Glucosidase from the Termite Nasutitermes Takasagoensis10.1128/AEM.07718-11 ↗
2. Uchiyama, T., Yaoi, K., and Miyazaki, K. (2015). Glucose-Tolerant β-Glucosidase Retrieved from a Kusaya Gravy Metagenome. Front. Microbiol., 6.10.3389/fmicb.2015.00548 ↗
3. Saccharification of Agricultural Residues10.1021/ie50421a006 ↗
4. Sustainable Valorization of Sugar Industry Waste: Status, Opportunities, and Challenges10.1016/j.biortech.2020.122929 ↗
5. Production of Multiple Xylanolytic and Cellulolytic Enzymes by Thermophilic Fungus Myceliophthora sp. IMI 38709910.1016/j.biortech.2006.02.009 ↗
6. Comparison of SHF and SSF processes from sugar cane bagasse for ethanol production by Saccharomyces cerevisiae10.1590/S0100-40422010000400027 ↗
7. Bioconversion of Lignocellulosic Biomass: Biochemical and Molecular Perspectives10.1007/s10295-008-0327-8 ↗
8. β-Glucosidases10.1007/s00018-010-0399-2 ↗
9. Selecting β-Glucosidases to Support Cellulases in Cellulose Saccharification10.1186/1754-6834-6-105 ↗
10. A Mechanism of Glucose Tolerance and Stimulation of GH1 β-Glucosidases10.1038/srep17296 ↗
11. Glucose Tolerant and Glucose Stimulated β-Glucosidases—A Review10.1016/j.biortech.2018.07.137 ↗
12. Zhao, L., Pang, Q., Xie, J., Pei, J., Wang, F., and Fan, S. (2013). Enzymatic Properties of Thermoanaerobacterium Thermosaccharolyticum β-Glucosidase Fused to Clostridium Cellulovorans Cellulose Binding Domain and Its Application in Hydrolysis of Microcrystalline Cellulose. BMC Biotechnol., 13.10.1186/1472-6750-13-101 ↗
13. Secretory Expression, Characterization and Docking Study of Glucose-Tolerant β-Glucosidase from B. Subtilis10.1016/j.ijbiomac.2016.01.001 ↗
14. Study of the Enzymatic Hydrolysis of Cellulose for Production of Fuel Ethanol by the Simultaneous Saccharification and Fermentation Process10.1002/bit.260410903 ↗
15. Ouyang, B., Wang, G., Zhang, N., Zuo, J., Huang, Y., and Zhao, X. (2023). Recent Advances in β-Glucosidase Sequence and Structure Engineering: A Brief Review. Molecules, 28.10.3390/molecules28134990 ↗
16. Thermostable Cellulose Saccharifying Microbial Enzymes: Characteristics, Recent Advances and Biotechnological Applications10.1016/j.ijbiomac.2021.08.024 ↗
17. Thermostabilizing Mechanisms of Canonical Single Amino Acid Substitutions at a GH1 β-Glucosidase Probed by Multiple MD and Computational Approaches
18. Wet Oxidation Pretreatment, Enzymatic Hydrolysis and Simultaneous Saccharification and Fermentation of Clover–Ryegrass Mixtures10.1016/j.biortech.2008.04.039 ↗
19. A Short Review on SSF–an Interesting Process Option for Ethanol Production from Lignocellulosic Feedstocks10.1186/1754-6834-1-7 ↗
20. Cloning with Kinetic and Thermodynamic Insight of a Novel Hyperthermostable β-Glucosidase from Thermotoga Naphthophila RKU-10T with Excellent Glucose Tolerance10.1016/j.molcatb.2015.12.005 ↗
21. A Novel Thermophilic β-Glucosidase from Caldicellulosiruptor Bescii: Characterization and Its Synergistic Catalysis with Other Cellulases10.1016/j.molcatb.2012.09.016 ↗
22. Genes Encoding Two Different Beta-Glucosidases of Thermoanaerobacter Brockii Are Clustered in a Common Operon10.1128/aem.63.10.3902-3910.1997 ↗
23. Comparative Analysis of Three Hyperthermophilic GH1 and GH3 Family Members with Industrial Potential10.1016/j.nbt.2014.07.009 ↗
24. Highly Accurate Protein Structure Prediction with AlphaFold10.1038/s41586-021-03819-2 ↗
25. Characterization of Glucose-Tolerant β-Glucosidases Used in Biofuel Production under the Bioinformatics Perspective: A Systematic Review
26. Mariano, D., Pantuza, N., Santos, L.H., Rocha, R.E.O., de Lima, L.H.F., Bleicher, L., and de Melo-Minardi, R.C. (2020). Glutantβase: A Database for Improving the Rational Design of Glucose-Tolerant β-Glucosidases. BMC Mol. Cell Biol., 21.10.1186/s12860-020-00293-y ↗
27. Costa, L.S.C., Mariano, D.C.B., Rocha, R.E.O., Kraml, J., da Silveira, C.H., Liedl, K.R., de Melo-Minardi, R.C., and de Lima, L.H.F. (2019). Molecular Dynamics Gives New Insights into the Glucose Tolerance and Inhibition Mechanisms on β-Glucosidases. Molecules, 24.10.3390/molecules24183215 ↗
28. Conformational Flexibility Correlates with Glucose Tolerance for Point Mutations in β-Glucosidases—A Computational Study10.1080/07391102.2020.1734484 ↗
29. Mariano, D., Santos, L.H., Machado, K.D.S., Werhli, A.V., de Lima, L.H.F., and de Melo-Minardi, R.C. (2019). A Computational Method to Propose Mutations in Enzymes Based on Structural Signature Variation (SSV). Int. J. Mol. Sci., 20.10.3390/ijms20020333 ↗
30. Protein Cutoff Scanning: A Comparative Analysis of Cutoff Dependent and Cutoff Free Methods for Prospecting Contacts in Proteins10.1002/prot.22187 ↗
31. Pires, D.E., de Melo-Minardi, R.C., dos Santos, M.A., da Silveira, C.H., Santoro, M.M., and Meira, W. (2011). Cutoff Scanning Matrix (CSM): Structural Classification and Function Prediction by Protein Inter-Residue Distance Patterns. BMC Genom., 12.10.1186/1471-2164-12-S4-S12 ↗
32. Rational Protein Design for Thermostabilization of Glycoside Hydrolases Based on Structural Analysis10.1007/s00253-018-9288-7 ↗
33. Sharma, S., Vaid, S., Bhat, B., Singh, S., and Bajaj, B.K. (2019). Thermostable Enzymes for Industrial Biotechnology. Advances in Enzyme Technology, Elsevier.10.1016/B978-0-444-64114-4.00017-0 ↗
34. In Silico Approach to Elucidate Factors Associated with GH1 β-Glucosidase Thermostability10.22207/JPAM.13.4.07 ↗
35. Thermal Stability Enhancement: Fundamental Concepts of Protein Engineering Strategies to Manipulate the Flexible Structure10.1016/j.ijbiomac.2022.06.154 ↗
36. Sourcing Thermotolerant Poly(Ethylene Terephthalate) Hydrolase Scaffolds from Natural Diversity10.1038/s41467-022-35237-x ↗
37. UniProt: A Hub for Protein Information10.1093/nar/gku989 ↗
38. AlphaFold Protein Structure Database in 2024: Providing Structure Coverage for over 214 Million Protein Sequences10.1093/nar/gkad1011 ↗
39. aCSM: Noise-Free Graph-Based Signatures to Large-Scale Receptor-Based Ligand Prediction10.1093/bioinformatics/btt058 ↗
40. Martins, P., Mariano, D., Carvalho, F.C., Bastos, L.L., Moraes, L., Paixão, V., and Cardoso de Melo-Minardi, R. (2023). Propedia v2.3: A Novel Representation Approach for the Peptide-Protein Interaction Database Using Graph-Based Structural Signatures. Front. Bioinform., 3.10.3389/fbinf.2023.1103103 ↗
41. Demšar, J., Zupan, B., Leban, G., and Curk, T. (2004). Orange: From Experimental Machine Learning to Interactive Data Mining, Springer.10.1007/978-3-540-30116-5_58 ↗
42. TemStaPro: Protein Thermostability Prediction Using Sequence Representations from Protein Language Models10.1093/bioinformatics/btae157 ↗
43. Clustal Omega, Accurate Alignment of Very Large Numbers of Sequences10.1007/978-1-62703-646-7_6 ↗
44. Schrödinger, L. (2019). The PyMOL Molecular Graphics System, V2.0.0, Schrödinger, LLC.
45. Improvements to the APBS Biomolecular Solvation Software Suite10.1002/pro.3280 ↗
46. Web Servers and Services for Electrostatics Calculations with APBS and PDB2PQR10.1002/jcc.21720 ↗
47. PDB2PQR: An Automated Pipeline for the Setup of Poisson–Boltzmann Electrostatics Calculations10.1093/nar/gkh381 ↗
48. PDB2PQR: Expanding and Upgrading Automated Preparation of Biomolecular Structures for Molecular Simulations10.1093/nar/gkm276 ↗
49. CABS-Flex 2.0: A Web Server for Fast Simulations of Flexibility of Protein Structures10.1093/nar/gky356 ↗
50. CABS-Flex: Server for Fast Simulation of Protein Structure Fluctuations10.1093/nar/gkt332 ↗
51. The Carbohydrate-Active EnZymes Database (CAZy): An Expert Resource for Glycogenomics10.1093/nar/gkn663 ↗
52. Accurate Prediction of Enzyme Thermostabilization with Rosetta Using AlphaFold Ensembles10.1021/acs.jcim.2c01083 ↗
53. Protein Rigidity and Thermophilic Adaptation10.1002/prot.22946 ↗
54. Protein Stability and Enzyme Activity at Extreme Biological Temperatures10.1088/0953-8984/22/32/323101 ↗