A novel fungal metal-dependent alpha-l-arabinofuranosidase of family 54 glycoside hydrolase shows expanded substrate specificity

Autor(es)

Leal Motta, Maria Lorenza ^[1] ; Ferreira Filho, Jaire Alves ^[1] ; de Melo, Ricardo Rodrigues ^[2] ; Zanphorlin, Leticia Maria ^[2] ; dos Santos, Clelton Aparecido ^[2] ; de Souza, Anete Pereira ^[1]

Afiliação do(s) autor(es)

^[1] Univ Campinas UNICAMP, Ctr Mol Biol & Genet Engn CBMEG, Dept Biol Vegetal, BR-13083875 Campinas, SP - Brazil

^[2] Brazilian Ctr Res Energy & Mat CNPEM, Brazilian Biorenewables Natl Lab LNBR, BR-13083100 Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 11, n. 1 MAY 26 2021.
• Citações Web of Science: 0

Resumo

Trichoderma genus fungi present great potential for the production of carbohydrate-active enzymes (CAZYmes), including glycoside hydrolase (GH) family members. From a renewability perspective, CAZYmes can be biotechnologically exploited to convert plant biomass into free sugars for the production of advanced biofuels and other high-value chemicals. GH54 is an attractive enzyme family for biotechnological applications because many GH54 enzymes are bifunctional. Thus, GH54 enzymes are interesting targets in the search for new enzymes for use in industrial processes such as plant biomass conversion. Herein, a novel metal-dependent GH54 arabinofuranosidase (ThABF) from the cellulolytic fungus Trichoderma harzianum was identified and biochemically characterized. Initial in silico searches were performed to identify the GH54 sequence. Next, the gene was cloned and heterologously overexpressed in Escherichia coli. The recombinant protein was purified, and the enzyme's biochemical and biophysical properties were assessed. GH54 members show wide functional diversity and specifically remove plant cell substitutions including arabinose and galactose in the presence of a metallic cofactor. Plant cell wall substitution has a major impact on lignocellulosic substrate conversion into high-value chemicals. These results expand the known functional diversity of the GH54 family, showing the potential of a novel arabinofuranosidase for plant biomass degradation. (AU)

• Processo FAPESP: 17/14253-9 - Avaliação do potencial biotecnológico do sistema celulolítico de Xanthomonas axonopodis pv. citri: uma abordagem estrutural, funcional e aplicada
• Beneficiário: Ricardo Rodrigues de Melo
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 15/09202-0 - Estudo de regiões genômicas de Trichoderma harzianum associadas ao controle da expressão de enzimas envolvidas na degradação de biomassa
• Beneficiário: Anete Pereira de Souza
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 18/19660-4 - Elucidação de mecanismos genéticos envolvidos na hidrólise de biomassa vegetal por meio de genômica funcional
• Beneficiário: Anete Pereira de Souza
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 16/19775-0 - Desenvolvimento / suplementação de coquetéis enzimáticos destinados à hidrólise de biomassa vegetal: utilização de mutagênese sítio-dirigida e proteínas quiméricas
• Beneficiário: Clelton Aparecido dos Santos
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 19/08855-1 - Novos mecanismos de P450: uma estratégia enzimática para a produção de hidrocarbonetos renováveis
• Beneficiário: Leticia Maria Zanphorlin
• Modalidade de apoio: Auxílio à Pesquisa - Regular