Applied Biochemistry and Biotechnology

, Volume 188, Issue 2, pp 395–409 | Cite as

Improved Thermal and Reusability Properties of Xylanase by Genipin Cross-Linking to Magnetic Chitosan Particles

  • Jorge Gracida
  • Teresita Arredondo-Ochoa
  • Blanca E. García-Almendárez
  • Monserrat Escamilla-García
  • Keiko Shirai
  • Carlos Regalado
  • Aldo Amaro-ReyesEmail author


Enzymes are gradually increasingly preferred over chemical processes, but commercial enzyme applications remain limited due to their low stability and low product recovery, so the application of an immobilization technique is required for repeated use. The aims of this work were to produce stable enzyme complexes of cross-linked xylanase on magnetic chitosan, to describe some characteristics of these complexes, and to evaluate the thermal stability of the immobilized enzyme and its reusability. A xylanase was cross-linked to magnetite particles prepared by in situ co-precipitation of iron salts in a chitosan template. The effect of temperature, pH, kinetic parameters, and reusability on free and immobilized xylanase was evaluated. Magnetization, morphology, size, structural change, and thermal behavior of immobilized enzyme were described. 1.0 ± 0.1 μg of xylanase was immobilized per milligram of superparamagnetic chitosan nanoparticles via covalent bonds formed with genipin. Immobilized xylanase showed thermal, pH, and catalytic velocity improvement compared to the free enzyme and can be reused three times. Heterogeneous aggregates of 254 nm were obtained after enzyme immobilization. The immobilization protocol used in this work was successful in retaining enzyme thermal stability and could be important in using natural compounds such as Fe3O4@Chitosan@Xylanase in the harsh temperature condition of relevant industries.


Xylanase Immobilization Cross-linking Magnetic nanoparticles Chitosan 



This work was supported by the Consejo Nacional de Ciencia y Tecnología, Mexico [CB-2014-241208].

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Facultad de QuímicaUniversidad Autónoma de Querétaro, Centro UniversitarioQuerétaroMexico
  2. 2.DIPA, PROPAC, Facultad de QuímicaUniversidad Autónoma de Querétaro, Centro UniversitarioQuerétaroMexico
  3. 3.Departmento de BiotecnologíaUniversidad Autonoma MetropolitanaMexico CityMexico

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