Skip to main content
SpringerLink
Log in

Immobilized Laccase on Activated Poly(Vinyl Alcohol) Microspheres For Enzyme Thermistor Application

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Poly(vinyl alcohol) (PVA) microspheres were prepared by inverse suspension crosslinked method, with glutaraldehyde as a crosslinking agent. PVA microspheres activated with aldehyde groups were employed for Trametes versicolor laccase immobilization. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy were used to characterize the activated PVA microspheres and PVA microspheres with immobilized laccase (Lac/PVA microspheres), which show that laccase was successfully immobilized on the PVA microspheres. The optimum pH and temperature coupling conditions for the immobilized laccase were determined to be 3.3 and 30 °C, respectively. Residual activity was also investigated by soaking the immobilized laccase in organic solvents at different concentrations, proving it chemically stable. Immobilized laccase exhibited good storage stability at 4 °C. The enzyme biosensor showed good performance in 2,2-azinobis(3-ethylthiazoline-6-sulfonate) and bisphenol A, with concentration ranges of 2 to 8 mM and 0.05 to 0.25 mM, respectively. Therefore, PVA microspheres may have high potential as support for enzyme thermistor applications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  1. Kudanga, T., Nyanhongo, G. S., Guebitz, G. M., & Burton, S. (2011). Potential applications of laccase-mediated coupling and grafting reactions: a review. Enzyme and Microbial Technology, 48, 195–208.

    Article  CAS  Google Scholar 

  2. Tastan, E., Onder, S., & Kok, F. N. (2011). Immobilization of laccase on polymer grafted polytetrafluoroethylene membranes for biosensor construction. Talanta, 84, 524–530.

    Article  CAS  Google Scholar 

  3. Xie, B., Ramanathan, K., & Danielsson, B. (2000). Mini/micro thermal biosensors and other related devices for biochemical/clinical analysis and monitoring. Trac-Trends Analytical Chemistry, 19, 340–349.

    Article  CAS  Google Scholar 

  4. Ramanathan, K., Rank, M., Svitel, J., Dzgoev, A., & Danielsson, B. (1999). The development and applications of thermal biosensors for bioprocess monitoring. Trends in Biotechnology, 17, 499–505.

    Article  CAS  Google Scholar 

  5. Yakovleva, M., Bhand, S., & Danielsson, B. (2013). The enzyme thermistor—a realistic biosensor concept. A critical review. Analytica Chimica Acta, 766, 1–12.

    Article  CAS  Google Scholar 

  6. Pirvutoiu, S., Surugiu, I., Dey, E. S., Ciucu, A., Magearu, V., & Danielsson, B. (2001). Flow injection analysis of mercury(II) based on enzyme inhibition and thermometric detection. Analyst, 126, 1612–1616.

    Article  CAS  Google Scholar 

  7. Yakovleva, M., Buzas, O., Matsumura, H., Samejima, M., Igarashi, K., Larsson, P. O., Gorton, L., & Danielsson, B. (2012). A novel combined thermometric and amperometric biosensor for lactose determination based on immobilised cellobiose dehydrogenase. Biosensors & Bioelectronics, 31, 251–256.

    Article  CAS  Google Scholar 

  8. Salman, S., Soundararajan, S., Safina, G., Satoh, I., & Danielsson, B. (2008). Hydroxyapatite as a novel reversible in situ adsorption matrix for enzyme thermistor-based FIA. Talanta, 77, 490–493.

    Article  CAS  Google Scholar 

  9. Ramanathan, K., Jonsson, B. R., & Danielsson, B. (2001). Sol-gel based thermal biosensor for glucose. Analytica Chimica Acta, 427, 1–10.

    Article  CAS  Google Scholar 

  10. Kim, H. J., Suma, Y., Lee, S. H., Kim, J. A., & Kim, H. S. (2012). Immobilization of horseradish peroxidase onto clay minerals using soil organic matter for phenol removal. Journal of Molecular Catalysis B: Enzymatic, 83, 8–15.

    Article  CAS  Google Scholar 

  11. Prodanovic, O., Prokopijevic, M., Spasojevic, D., Stojanovic, Z., Radotic, K., Knezevic-Jugovic, Z. D., & Prodanovic, R. (2012). Improved covalent immobilization of horseradish peroxidase on macroporous glycidyl methacrylate-based copolymers. Applied Biochemistry and Biotechnology, 168, 1288–1301.

    Article  CAS  Google Scholar 

  12. Olcer, Z., Ozmen, M. M., Sahin, Z. M., Yilmaz, F., & Tanriseven, A. (2013). Highly efficient method towards in situ immobilization of invertase using cryogelation. Applied Biochemistry and Biotechnology, 171, 2142–2152.

    Article  CAS  Google Scholar 

  13. Tsai, H. C., & Doong, R. A. (2007). Preparation and characterization of urease-encapsulated biosensors in poly(vinyl alcohol)-modified silica sol-gel materials. Biosensors & Bioelectronics, 23, 66–73.

    Article  CAS  Google Scholar 

  14. Bai, X., Ye, Z. F., Qu, Y. Z., Li, Y. F., & Wang, Z. Y. (2009). Immobilization of nanoscale Fe(0) in and on PVA microspheres for nitrobenzene reduction. Journal of Hazardous Materials, 172, 1357–1364.

    Article  CAS  Google Scholar 

  15. Kumar, J., & D'Souza, S. F. (2008). Preparation of PVA membrane for immobilization of GOD for glucose biosensor. Talanta, 75, 183–188.

    Article  CAS  Google Scholar 

  16. Derwinska, K., Sauer, U., & Preininger, C. (2008). Adsorption versus covalent, statistically oriented and covalent, site-specific IgG immobilization on poly(vinyl alcohol)-based surfaces. Talanta, 77, 652–658.

    Article  CAS  Google Scholar 

  17. Bryjak, J., Kruczkiewicz, P., Rekuc, A., & Peczynska-Czoch, W. (2007). Laccase immobilization on copolymer of butyl acrylate and ethylene glycol dimethacrylate. Biochemical Engineering Journal, 35, 325–332.

    Article  CAS  Google Scholar 

  18. Mendes, A. A., Giordano, R. C., Giordano, R. D. L. C., & de Castro, H. F. (2011). Immobilization and stabilization of microbial lipases by multipoint covalent attachment on aldehyde-resin affinity: application of the biocatalysts in biodiesel synthesis. Journal of Molecular Catalysis B: Enzymatic, 68, 109–115.

    Article  CAS  Google Scholar 

  19. Zhu, Y., Kaskel, S., Shi, J., Wage, T., & van Pée, K. H. (2007). Immobilization of Trametes versicolor laccase on magnetically separable mesoporous silica spheres. Chemistry of Materials, 19, 6408–6413.

    Article  CAS  Google Scholar 

  20. Arica, M. Y. (2000). Immobilization of polyphenol oxidase on carboxymethylcellulose hydrogel beads: preparation and characterization. Polymer International, 49, 775–781.

    Article  CAS  Google Scholar 

  21. Filho, M., Pessela, B. C., Mateo, C., Carrascosa, A. V., Fernandez-Lafuente, R., & Guisan, J. M. (2008). Reversible immobilization of a hexameric alpha-galactosidase from Thermus sp strain T2 on polymeric ionic exchangers. Process Biochemistry, 43, 1142–1146.

    Article  CAS  Google Scholar 

  22. Zaks, A., & Klibanov, A. M. (1988). The effect of water on enzyme action in organic media. Journal of Biological Chemistry, 263, 8017–8021.

    CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge the support provided by the Natural Science Foundation of Jiangsu Province of China (Grant No. BK20130828), the Specialized Research Fund for the Doctoral Program of Higher Education (Grant No. 20130094120009), the National Natural Science Foundation of China (Grant No. 51308183), and the Fundamental Research Funds for the Central Universities (Grant No. 2013B32214).

Author information

Authors and Affiliations

  1. Key Laboratory of Integrated Regulation and Resource Development on Shallow Lake of Ministry of Education, College of Environment, Hohai University, Nanjing, 210098, China

    Xue Bai, Haixin Gu, Bei Yang, Xin Huang & Qi Zhang

  2. State Key Joint Laboratory of Environmental Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China

    Xue Bai & Hanchang Shi

  3. State Key Laboratory of Explosion Science and Technology, Beijing Institutive of Technology, Beijing, 100081, China

    Wei Chen

Authors
  1. Xue Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Haixin Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hanchang Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bei Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xin Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Qi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xue Bai.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bai, X., Gu, H., Chen, W. et al. Immobilized Laccase on Activated Poly(Vinyl Alcohol) Microspheres For Enzyme Thermistor Application. Appl Biochem Biotechnol 173, 1097–1107 (2014). https://doi.org/10.1007/s12010-014-0913-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-014-0913-3

Keywords

Search

Navigation