High-resolution structure of a modular hyperthermostable endo-beta-1,4-mannanase from Thermotoga petrophila: The ancillary immunoglobulin-like module is a thermostabilizing domain

Author(s)

da Silva, Viviam M. ^{[1, 2]} ; Cabral, Aline D. ^[3] ; Speranca, Marcia A. ^[3] ; Squina, Fabio M. ^[4] ; Muniz, Joao Renato C. ^[5] ; Martin, Lydie ^[2] ; Nicolet, Yvain ^[2] ; Garcia, Wanius ^[1]

Affiliation

^[1] Univ Fed ABC UFABC, Ctr Ciencias Nat & Humanas, Santo Andre, SP - Brazil

^[2] Univ Grenoble Alpes, CEA, CNRS, IBS, Metalloprot Unit, F-38000 Grenoble - France

^[3] Univ Fed ABC UFABC, Ctr Ciencias Nat & Humanas, Sao Bernardo Do Campo, SP - Brazil

^[4] Univ Sorocaba UNISO, Programa Proc Tecnol & Ambientais, Sorocaba, SP - Brazil

^[5] Univ Sao Paulo, Sao Carlos Inst Phys IFSC, Sao Carlos, SP - Brazil

• Document type: Journal article
• Source: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1868, n. 8 AUG 2020.
• Web of Science Citations: 0

Abstract

The endo-beta-1,4-mannanase from the hyperthermostable bacterium Thermotoga petrophila (TpMan) is an enzyme that catalyzes the hydrolysis of mannan and heteromannan polysaccharides. Of the three domains that comprise TpMan, the N-terminal GH5 catalytic domain and the C-terminal carbohydrate-binding domain are connected through a central ancillary domain of unknown structure and function. In this study, we report the partial crystal structure of the TpMan at 1.45 angstrom resolution, so far, the first modular hyperthermostable endo-beta-1,4-mannanase structure determined. The structure exhibits two domains, a (beta/alpha)(8)-barrel GH5 catalytic domain connected via a linker to the central domain with an immunoglobulin-like beta-sandwich fold formed of seven beta-strands. Functional analysis showed that whereas the immunoglobulin-like domain does not have the carbohydrate-binding function, it stacks on the GH5 catalytic domain acting as a thermostabilizing domain and allowing operation at hyperthermophilic conditions. The carbohydrate-binding domain is absent in the crystal structure most likely due to its high flexibility around the immunoglobulin-like domain which may act also as a pivot. These results represent new structural and functional information useful on biotechnological applications for biofuel and food industries. (AU)

• FAPESP's process: 13/26096-4 - Heterologous expression of the chitinase enzyme of Leishmania (L.) infantum chagasi, Leishmania (V.) braziliensis and Leishmania (V.) amazonensis: serological diagnosis and comparative molecular study using expression systems of insects cell and bacteria
• Grantee: Aline Diniz Cabral
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 16/15836-5 - Modular Hyperthermostable endo-beta-1,4-mannanase: determination of the three-dimensional structure of the central domain and its role in the thermostabilization of whole enzyme
• Grantee: Viviam Moura da Silva
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 17/17275-3 - Studies of the mode of action of two lytic polysaccharide monooxygenases from insect (order: isoptera): molecular structure, bioinorganic chemistry and biotechnological applications
• Grantee: Wanius José Garcia da Silva
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/16291-5 - Structural and biophysical characterization of thermophilic enzymes prospected from the fungus Thielavia terrestris for biomass degradation and biotechnological products generation
• Grantee: João Renato Carvalho Muniz
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/02065-5 - Modular Hyperthermostable endo-beta-1,4-mannanase: determination of the three-dimensional structure of the Central Domain an its role in the thermostabilization of whole enzyme
• Grantee: Viviam Moura da Silva
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/14514-4 - Characterization of the chitinase from South American endemic Leishmania species: use in diagnosis in humans, dogs and sandflies.
• Grantee: Marcia Aparecida Speranca
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/50897-0 - INCT 2014: Open-acess Medicinal Chemistry Centre (OpenMedChem)
• Grantee: Katlin Brauer Massirer
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 15/50590-4 - Lignin valorization in cellulosic ethanol plants: biocatalytic conversion via ferulic acid to high value chemicals
• Grantee: Fábio Márcio Squina
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Thematic Grants