Autor(es):Mostrar menos -

Cota, Junio ^{[1, 2]} ; Oliveira, Leandro C. ^{[2, 3]} ; Damasio, Andre R. L. ^[2] ; Citadini, Ana P. ^[2] ; Hoffmam, Zaira B. ^[2] ; Alvarez, Thabata M. ^[2] ; Codima, Carla A. ^[2] ; Leite, Vitor B. P. ^[3] ; Pastore, Glaucia ^[4] ; de Oliveira-Neto, Mario ^[1] ; Murakami, Mario T. ^[5] ; Ruller, Roberto ^[2] ; Squina, Fabio M. ^[2]

Afiliação do(s) autor(es)

^[1] UNESP, Inst Biociencias, Dept Fis & Biofis, Botucatu, SP - Brazil

^[2] Lab Nacl Ciencia & Tecnol Bioetanol CTBE CNPEM, Campinas, SP - Brazil

^[3] Univ Estadual Paulista, UNESP, IBILCE, Dept Fis, Sao Jose Do Rio Preto, SP - Brazil

^[4] Univ Estadual Campinas, Fac Engn Alimentos, Campinas, SP - Brazil

^[5] Lab Nacl Biociencias LNBio CNPEM, Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS; v. 1834, n. 8, p. 1492-1500, AUG 2013.
• Citações Web of Science: 15

Resumo

Multifunctional enzyme engineering can improve enzyme cocktails for emerging biofuel technology. Molecular dynamics through structure-based models (SB) is an effective tool for assessing the tridimensional arrangement of chimeric enzymes as well as for inferring the functional practicability before experimental validation. This study describes the computational design of a bifunctional xylanase-lichenase chimera (XylLich) using the xynA and bglS genes from Bacillus sub fills. In silico analysis of the average solvent accessible surface area (SAS) and the root mean square fluctuation (RMSF) predicted a fully functional chimera, with minor fluctuations and variations along the polypeptide chains. Afterwards, the chimeric enzyme was built by fusing the xynA and bglS genes. XylLich was evaluated through small-angle X-ray scattering (SAXS) experiments, resulting in scattering curves with a very accurate fit to the theoretical protein model. The chimera preserved the biochemical characteristics of the parental enzymes, with the exception of a slight variation in the temperature of operation and the catalytic efficiency (k(cat)/K-m). The absence of substantial shifts in the catalytic mode of operation was also verified. Furthermore, the production of chimeric enzymes could be more profitable than producing a single enzyme separately, based on comparing the recombinant protein production yield and the hydrolytic activity achieved for XylLich with that of the parental enzymes. (C) 2013 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 11/13242-7 - Estudos de modelos de mecanismos enzimáticos para o aprimoramento da produção de biocombustíveis
• Beneficiário: Leandro Cristante de Oliveira
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 11/02169-7 - Aspergillus nidulans como modelo para expressão heteróloga de celulases e hemicelulases
• Beneficiário: André Ricardo de Lima Damasio
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 08/58037-9 - Geração de biblioteca para conversão enzimática de biomassa a partir de metagenoma do solo
• Beneficiário: Fábio Márcio Squina
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Jovens Pesquisadores