Structural insights into the hydrolysis pattern and molecular dynamics simulations of GH45 subfamily a endoglucanase from Neurospora crassa OR74A

Autor(es)

Seiki Kadowaki, Marco Antonio ^[1] ; Polikarpov, Igor ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sao Carlos Inst Phys, Sao Carlos, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biochimie; v. 165, p. 275-284, OCT 2019.
• Citações Web of Science: 0

Resumo

Glycoside hydrolase (GH) family 45 is one of the smallest and poorly studied endoglucanase family with a broad biotechnological application ranging from treatment of textiles to conversion of complex cell wall polysaccharides into simple oligo- and monosaccharides. In a present study, GH45 cellulase from Neurospora crassa OR74A (NcCel45A) was characterized both biochemically and structurally. HPLC analysis of the hydrolytic products confirmed the endo-beta(1,4) mode of action of the enzyme. Moreover, such pattern revealed that NcCel45A cannot hydrolyze efficiently oligosaccharides with a degree of polymerization smaller than six. The crystal structure of NcCel45A catalytic domain in the apo-form was determined at 1.9 angstrom resolution and the structure of the enzyme bound to cellobiose was solved and refined to 1.8 angstrom resolution. Comparative structural analyses and molecular dynamics simulations show that the enzyme dynamics is affected by substrate binding. Taken together, MD simulations and statistical coupling analysis revealed previously unknown correlation of a loop 6 with the breakdown of cellulose substrates by GH45. (C) 2019 Published by Elsevier B.V. (AU)

• Processo FAPESP: 15/13684-0 - Estudos estruturais e funcionais de enzimas que participam na síntese e degradação de carboidratos complexos
• Beneficiário: Igor Polikarpov
• Linha de fomento: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/11730-3 - Estudos estruturais e funcionais de xilanases com potencial aplicação biotecnológica
• Beneficiário: Ana Gabriela Veiga Sepulchro
• Linha de fomento: Bolsas no Brasil - Iniciação Científica