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Purification and characterization studies of a thermostable β-xylanase from Aspergillus awamori

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Journal of Industrial Microbiology & Biotechnology

Abstract

This study presents data on the production, purification, and properties of a thermostable β-xylanase produced by an Aspergillus awamori 2B.361 U2/1 submerged culture using wheat bran as carbon source. Fractionation of the culture filtrate by membrane ultrafiltration followed by Sephacryl S-200 and Q-Sepharose chromatography allowed for the isolation of a homogeneous xylanase (PXII-1), which was 32.87 kDa according to MS analysis. The enzyme-specific activity towards soluble oat spelt xylan, which was found to be 490 IU/mg under optimum reaction conditions (50°C and pH 5.0–5.5), was 17-fold higher than that measured in the culture supernatant. Xylan reaction products were identified as xylobiose, xylotriose, and xylotetraose. K m values (mg ml−1) for soluble oat spelt and birchwood xylan were 11.8 and 9.45, respectively. Although PXII-1 showed 85% activity retention upon incubation at 50°C and pH 5.0 for 20 days, incubation at pH 7.0 resulted in 50% activity loss within 3 days. PXII-1 stability at pH 7.0 was improved in the presence of 20 mM cysteine, which allowed for 85% activity retention for 25 days. This study on the production in high yields of a remarkably thermostable xylanase is of significance due to the central role that this class of biocatalyst shares, along with cellulases, for the much needed enzymatic hydrolysis of biomass. Furthermore, stable xylanases are important for the manufacture of paper, animal feed, and xylooligosaccharides.

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Acknowledgments

This work was supported by research and scholarship grants from the National Council for Scientific and Technological Development (CNPq) of the Brazilian Ministry of Science and Technology and from a scholarship grant from the Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES) of the Ministry of Education. We are thankful to Dr. Marília Martins Nishikawa from the National Institute of Quality Control in Health (INCQS) of the Oswaldo Cruz Foundation for preserving the Aspergillus awamori strain.

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Authors and Affiliations

  1. Laboratory of Enzyme Technology, Biochemistry Department, Chemistry Institute, Federal University of Rio de Janeiro, Av. Athos da Silveira Ramos 149, Bloco A, Ilha do Fundão, CEP 21941-909, Rio de Janeiro, RJ, Brazil

    Ricardo Sposina Sobral Teixeira & Elba Pinto da Silva Bon

  2. Laboratory of Enzymology, Department of Cellular Biology, University of Brasilia, CEP 70910-900, Asa Norte, Brasilia, DF, Brazil

    Félix Gonçalves Siqueira & Edivaldo Ximenes Ferreira Filho

  3. Laboratory of Biochemistry and Protein Chemistry, Department of Cellular Biology, University of Brasilia, CEP 70910-900, Asa Norte, Brasilia, DF, Brazil

    Marcelo Valle de Souza

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  1. Ricardo Sposina Sobral Teixeira
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Correspondence to Elba Pinto da Silva Bon.

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Teixeira, R.S.S., Siqueira, F.G., Souza, M.V. et al. Purification and characterization studies of a thermostable β-xylanase from Aspergillus awamori . J Ind Microbiol Biotechnol 37, 1041–1051 (2010). https://doi.org/10.1007/s10295-010-0751-4

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