Impact of cellulose properties on enzymatic degradation by bacterial GH48 enzymes: Structural and mechanistic insights from processive Bacillus licheniformis Cel48B cellulase

Autor(es)

Araujo, Evandro A. ; Dias, Artur Hermano Sampaio ; Kadowaki, Marco A. S. ; Piyadov, Vasily ; Pellegrini, Vanessa O. A. ; Urio, Mateus B. ; Ramos, Luiz P. ; Skaf, Munir S. ; Polikarpov, Igor

• Tipo de documento: Artigo Científico
• Fonte: Carbohydrate Polymers; v. 264, p. 13-pg., 2021-04-12.

Resumo

Processive cellulases are highly efficient molecular engines involved in the cellulose breakdown process. However, the mechanism that processive bacterial enzymes utilize to recruit and retain cellulose strands in the catalytic site remains poorly understood. Here, integrated enzymatic assays, protein crystallography and computational approaches were combined to study the enzymatic properties of the processive BlCel48B cellulase from Bacillus licheniformis. Hydrolytic efficiency, substrate binding affinity, cleavage patterns, and the apparent processivity of bacterial BlCel48B are significantly impacted by the cellulose size and its surface morphology. BlCel48B crystallographic structure was solved with ligands spanning -5 to -2 and +1 to +2 subsites. Statistical coupling analysis and molecular dynamics show that co-evolved residues on active site are critical for stabilizing ligands in the catalytic tunnel. Our results provide mechanistic insights into BlCel48B molecular-level determinants of activity, substrate binding, and processivity on insoluble cellulose, thus shedding light on structure-activity correlations of GH48 family members in general. (AU)

• Processo FAPESP: 14/00769-5 - Estudos estruturais de hidrolases de glicosídeos em solução usando técnicas de espalhamento a baixo ângulo (SAS)
• Beneficiário: Vasilii Piiadov
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 13/08293-7 - CECC - Centro de Engenharia e Ciências Computacionais
• Beneficiário: Munir Salomao Skaf
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 15/13684-0 - Estudos estruturais e funcionais de enzimas que participam na síntese e degradação de carboidratos complexos
• Beneficiário: Igor Polikarpov
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 19/17350-0 - Modelagem molecular de enzimas ativas sobre carboidratos
• Beneficiário: Artur Hermano Sampaio Dias
• Modalidade de apoio: Bolsas no Brasil - Doutorado