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Purification and characterization of two xylanases from alkalophilic and thermophilic Bacillus licheniformis 77-2

  • Session 1B: Enzyme Catalysis and Engineering
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Abstract

The alkalophilic bacteria Bacillus licheniformis 77-2 produces significant quantities of thermostable cellulase-free xylanases. The crude xylanase was purified to apparent homogeneity by gel filtration (G-75) and ionic exchange chromatography (carboxymethyl sephadex, Q sepharose, and Mono Q), resulting in the isolation of two xylanases. The molecular masses of the enzymes were estimated to be 17 kDa (X-I) and 40 kDa (X-II), as determined by SDS-PAGE. The K m and V max values were 1.8 mg/mL and 7.05 U/mg protein (X-I), and 1.05 mg/mL and 9.1 U/mg protein (X-II). The xylanases demonstrated optimum activity at pH 7.0 and 8.0–10.0 for xylanase X-I and X-II, respectively, and, retained more than 75% of hydrolytic activity up to pH 11.0. The purified enzymes were most active at 70 and 75°C for X-I and X-II, respectively, and, retained more than 90% of hydrolytic activity after 1 h of heating at 50°C and 60°C for X-I and X-II, respectively. The predominant products of xylan hydrolysates indicated that these enzymes were endoxylanases.

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Authors and Affiliations

  1. Biochemistry and Applied Microbiology Laboratory, UNESP-State University of São Paulo, Rua Cristóvão Colombo n 2265, 15054-000, São José do Rio Preto, SP, Brazil

    Valquiria B. Damiano, Eleni Gomes, Heloiza Ferreira Alves-Prado & Roberto Da Silva

  2. Biology Institute, UNESP-State University of São Paulo, Rio Claro, SP

    Valquiria B. Damiano & Heloiza Ferreira Alves-Prado

  3. Chemistry Institute, USP-University of São Paulo, Ribeirão Preto, SP

    Richard Ward

Authors
  1. Valquiria B. Damiano
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  2. Richard Ward
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  3. Eleni Gomes
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  4. Heloiza Ferreira Alves-Prado
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  5. Roberto Da Silva
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Correspondence to Roberto Da Silva.

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Damiano, V.B., Ward, R., Gomes, E. et al. Purification and characterization of two xylanases from alkalophilic and thermophilic Bacillus licheniformis 77-2. Appl Biochem Biotechnol 129, 289–302 (2006). https://doi.org/10.1385/ABAB:129:1:289

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