GH11 xylanase from Aspergillus tamarii Kita: Purification by one-step chromatography and xylooligosaccharides hydrolysis monitored in real-time by mass spectrometry

Autor(es):Mostrar menos -

Heinen, P. R. ^[1] ; Bauermeister, A. ^[2] ; Ribeiro, L. F. ^[3] ; Messias, J. M. ^[2] ; Almeida, P. Z. ^[1] ; Moraes, L. A. B. ^[4] ; Vargas-Rechia, C. G. ^[2] ; de Oliveira, A. H. C. ^[4] ; Ward, R. J. ^[4] ; Filho, E. X. F. ^[5] ; Kadowaki, M. K. ^[6] ; Jorge, J. A. ^[7] ; Polizeli, M. L. T. M. ^[7]

Afiliação do(s) autor(es)

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

^[2] Univ Sao Paulo, Dept Fis & Quim, Fac Ciencias Farmaceut Ribeirao Preto, Ribeirao Preto, SP - Brazil

^[3] Johns Hopkins Univ, Dept Chem & Biomol Engn, Baltimore, MD 21218 - USA

^[4] Univ Sao Paulo, Dept Quim, Fac Filosofia Ciencias & Letras Ribeirao Preto, Ribeirao Preto, SP - Brazil

^[5] Univ Brasilia, Dept Biol Celular, Brasilia, DF - Brazil

^[6] Univ Estadual Oeste Parana, Ctr Ciencias Med & Farmaceut, Cascavel, Parana - Brazil

^[7] Univ Sao Paulo, Dept Biol, Fac Filosofia Ciencias & Letras Ribeirao Preto, Av Bandeirantes 3900, BR-14040901 Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 108, p. 291-299, MAR 2018.
• Citações Web of Science: 9

Resumo

The present study describes the one-step purification and biochemical characterization of an endo-1,4-beta-xylanase from Aspergillus tamarii Kita. Extracellular xylanase was purified to homogeneity 7.43-fold through CM-cellulose. Enzyme molecular weight and pI were estimated to be 19.5 kDa and 8.5, respectively. The highest activity of the xylanase was obtained at 60 degrees C and it was active over a broad .pH range (4.0-9.0), with maximal activity at pH 5.5. The enzyme was thermostable at 50 degrees C, retaining more than 70% of its initial activity for 480min. The K-0.5 and V-max values on beechwood xylan were 8.13 mg/mL and 1,330.20 mu mol/min/mg of protein, respectively. The ions Ba2+ and Ni2+, and the compounds beta-mercaptoethanol and DTT enhanced xylanase activity, while the heavy metals (Co2+, Cu2+, Hg+, Pb2+ and Zn2+) strongly inhibited the enzyme, at 5 mM. Enzymatic hydrolysis of xylooligosaccharides monitored in real-time by mass spectrometer showed that the shortest xylooligosaccharide more efficiently hydrolyzed by A. tamarii Kita xylanase corresponded to xylopentaose. In agreement, HPLC analyzes did not detect xylopentaose among the hydrolysis products of xylan. Therefore, this novel GH11 endo-xylanase displays a series of physicochemical properties favorable to its application in the food, feed, pharmaceutical and paper industries. (C) 2017 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 10/52322-3 - Bioprospecção de fungos filamentosos produtores de holoenzimas com aplicação em biorefinaria
• Beneficiário: Maria de Lourdes Teixeira de Moraes Polizeli
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOTA - Regular