Structural model and functional properties of an exo-polygalacturonase from Neosartorya glabra

Autor(es)

Desagiacomo, Carla Cristina Villela ^[1] ; Alnoch, Robson Carlos ^[2] ; Pinheiro, Vanessa Elisa ^[1] ; Cereia, Mariana ^[2] ; Machado, Carla Botelho ^[3] ; Damasio, Andre ^[4] ; Augusto, Marlei Josiele ^[5] ; Pedersoli, Wellington ^[1] ; Silva, Roberto Nascimento ^[1] ; Polizeli, Maria de Lourdes Teixeira de Moraes ^{[2, 1]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Bioquim & Imunol, BR-14049900 Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Biol, BR-14040901 Ribeirao Preto, SP - Brazil

^[3] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol, BR-13083970 Campinas - Brazil

^[4] Univ Campinas UNICAMP, Inst Biol, BR-13083862 Campinas, SP - Brazil

^[5] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Patol & Med Legal, BR-14040901 Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 186, p. 909-918, SEP 1 2021.
• Citações Web of Science: 0

Resumo

A purified exo-polygalacturonase of Neosartorya glabra (EplNg) was successfully characterized. EplNg native presented 68.2 kDa, with 32% carbohydrate content. The deglycosylated form showed 46.3 kDa and isoelectric point of 5.4. The identity of EpIlNg was confirmed as an exo-polygalacturonase class I (EC 3.2.1.67) using mass spectrometry and Western-Blotting. Capillary electrophoresis indicated that only galactumnic acid was released by the action of EplNg on sodium polypectate, confirming an exoenzyme character. The structural model confers that EplNg has a core formed by twisted parallel beta-sheets structure. Among twelve putative cysteines, ten were predicted to form disulfide bridges. The catalytic triad predicted is composed of Asp(223), ASp(245), and Asp(246) aligned along with a distance in 4-5 angstrom, suggesting that EplNg probably does not perform the standard inverting catalytic mechanism described for the GH28 family. EplNg was active from 30 to 90 degrees C, with maximum activity at 65 degrees C, pH 5.0. The Km and Vmax determined using sodium polypectate were 6.9 mg.mL(-1) and Vmax 690 mu mol.min(-1).mg(-1), respectively. EplNg was active and stable over a wide range of pH values and temperatures, confirming the interesting properties EplNg and provide a basis for the development of the enzyme in different biotechnological processes. (AU)

• Processo FAPESP: 14/50884-5 - INCT 2014: Instituto Nacional de Ciência e Tecnologia do Bioetanol
• Beneficiário: Marcos Silveira Buckeridge
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 20/00081-4 - Desenvolvimento de estratégias para imobilização e co-imobilização de enzimas envolvidas na degradação de biomassa
• Beneficiário: Robson Carlos Alnoch
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 18/07522-6 - Desenvolvimento de tecnologias ecológicas para a hidrólise de biomassa e resíduos da indústria de celulose e papel
• Beneficiário: Maria de Lourdes Teixeira de Moraes Polizeli
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 17/22669-0 - N-glicosilação e secreção de enzimas em fungos filamentosos
• Beneficiário: André Ricardo de Lima Damasio
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Regular