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Purification and characterization of two sugarcane bagasse-absorbable thermophilic xylanases from the mesophilic Cellulomonas flavigena

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Journal of Industrial Microbiology & Biotechnology

Abstract

We report the purification and characterization of two thermophilic xylanases from the mesophilic bacteria Cellulomonas flavigena grown on sugarcane bagasse (SCB) as the only carbon source. Extracellular xylanase activity produced by C. flavigena was found both free in the culture supernatant and associated with residual SCB. To identify some of the molecules responsible for the xylanase activity in the substrate-bound fraction, residual SCB was treated with 3 M guanidine hydrochloride and then with 6 M urea. Further analysis of the eluted material led to the identification of two xylanases Xyl36 (36 kDa) and Xyl53 (53 kDa). The pI for Xyl36 was 5.0, while the pI for Xyl53 was 4.5. Xyl36 had a K m value of 1.95 mg/ml, while Xyl53 had a K m value of 0.78 mg/ml. In addition to SCB, Xyl36 and Xyl53 were also able to bind to insoluble oat spelt xylan and Avicel, as shown by substrate-binding assays. Xyl36 and Xyl53 showed optimal activity at pH 6.5, and at optimal temperature 65 and 55°C, respectively. Xyl36 and Xyl53 retained 24 and 35%, respectively, of their original activity after 8 h of incubation at their optimal temperature. As far as we know, this is the first study on the thermostability properties of purified xylanases from microorganisms belonging to the genus Cellulomonas.

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Acknowledgments

This paper is dedicated, for the valuable teachings, to the memory of Dr. Ignacio Magaña-Plaza. We would like to thank Carmen Fontaine for the technical assistance. The research was funded by the Departamento de Biotecnología y Bioingeniería, CINVESTAV México.

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Authors and Affiliations

  1. Departamento de Biotecnología y Bioingeniería, CINVESTAV, Av. Instituto Politécnico Nacional, 2508, Mexico D. F., CP 07360, Mexico

    Alejandro Santiago-Hernández, Jesús Vega-Estrada, María del Carmen Montes-Horcasitas & María Eugenia Hidalgo-Lara

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  1. Alejandro Santiago-Hernández
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  2. Jesús Vega-Estrada
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  3. María del Carmen Montes-Horcasitas
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  4. María Eugenia Hidalgo-Lara
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Correspondence to María Eugenia Hidalgo-Lara.

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Santiago-Hernández, A., Vega-Estrada, J., del Carmen Montes-Horcasitas, M. et al. Purification and characterization of two sugarcane bagasse-absorbable thermophilic xylanases from the mesophilic Cellulomonas flavigena . J Ind Microbiol Biotechnol 34, 331–338 (2007). https://doi.org/10.1007/s10295-006-0202-4

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