Electrostatic interaction optimization improves catalytic rates and thermotolerance on xylanases

Author(s)

Contessoto, Vinicius de Godoi ; Ramos, Felipe Cardoso ; de Melo, Ricardo Rodrigues ; de Oliveira, Vinicius Martins ; Scarpassa, Josiane Aniele ; de Sousa, Amanda Silva ; Zanphorlin, Leticia Maria ; Slade, Gabriel Gouvea ; Pereira Leite, Vitor Barbanti ; Ruller, Roberto

• Document type: Journal article
• Source: BIOPHYSICAL JOURNAL; v. 120, n. 11, p. 9-pg., 2021-06-01.

Abstract

Understanding the aspects that contribute to improving proteins' biochemical properties is of high relevance for protein engineering. Properties such as the catalytic rate, thermal stability, and thermal resistance are crucial for applying enzymes in the industry. Different interactions can influence those biochemical properties of an enzyme. Among them, the surface charge-charge interactions have been a target of particular attention. In this study, we employ the Tanford-Kirkwood solvent accessibility model using the Monte Carlo algorithm (TKSA-MC) to predict possible interactions that could improve stability and catalytic rate of a WT xylanase (XynA(WT)) and its M6 xylanase (XynA(M6)) mutant. The modeling prediction indicates that mutating from a lysine in position 99 to a glutamic acid (K99E) favors the native state stabilization in both xylanases. Our lab results showed that mutated xylanases had their thermotolerance and catalytic rate increased, which conferred higher processivity of delignified sugarcane bagasse. The TKSA-MC approach employed here is presented as an efficient computational-based design strategy that can be applied to improve the thermal resistance of enzymes with industrial and biotechnological applications. (AU)

• FAPESP's process: 16/19766-1 - Biological macromolecules energy landscapes with applications in biotechnology and in biomedicine
• Grantee: Vitor Barbanti Pereira Leite
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/13998-8 - Rational evolution by computational methods applied to predict mutations in enzymes to biofuels production
• Grantee: Vinícius de Godoi Contessoto
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 17/14253-9 - Evaluation of the biotechnological potential of the cellulolytic system of Xanthomonas axonopodis pv. citri: a structural, functional and applied approach
• Grantee: Ricardo Rodrigues de Melo
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 18/11614-3 - Effect of pH and Cancer-Activating Mutations on Functional Transition of Estrogen Receptor
• Grantee: Vinicius Martins de Oliveira
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/06862-7 - Computational studies in protein folding and enzymes engineering involved in bioethanol production
• Grantee: Vitor Barbanti Pereira Leite
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/22540-3 - Studies of energy landscapes of biological macromolecules
• Grantee: Vitor Barbanti Pereira Leite
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/09662-7 - Rational Evolution by Computational Methods Applied in Enzymes Related to Bioethanol Production
• Grantee: Vinícius de Godoi Contessoto
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor