Skip to main content
SpringerLink
Log in

Purification and Characterization of Class III Lipase from a White-Rot Fungus Pleurotus ostreatus

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

Abstract

Pleurotus ostreatus is an edible white-rot fungus with lignocellulosic biomass degrading enzymes that have been studied extensively. However, until now, lipolytic enzymes from P. ostreatus, which degrade extractives in lignocellulosic biomass, have not been purified and characterized. In this study, P. ostreatus was inoculated into the rapeseed oil containing culture to induce lipase. The lipase in the culture broth was successfully purified to homogeneity by chromatographic methods. The molecular weight of the purified lipase was 27 kDa, and its optimal pH and temperature were 5.0 and 30 °C, respectively. The purified lipase showed high activity with the substrates 4-methylumbelliferyl (4-MU) decanoate (C10:0) and 4-MU oleate (C18:1), and no activity with 4-MU acetate (C2:0) and 4-MU butyrate (C4:0). The amino acid sequences and substrate specificities of the purified lipase suggested that it belonged to class III. Kinetic parameters measurements (Km and Vmax) showed that 4-MU palmitate had a high affinity for the purified lipase, and it was the substrate most efficiently hydrolyzed by the purified lipase.

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

Similar content being viewed by others

Data Availability

The datasets generated and/or analyzed during the current study are available from the corresponding author upon reasonable request.

References

  1. Ghosh, P. K., Saxena, R. K., Gupta, R., Yadav, R. P., & Davidson, S. (1996). Microbial lipases: Production and applications. Science Progress, 79(Pt 2), 119–157.

    CAS  Google Scholar 

  2. Macrae, A., & Hammond, R. (1985). Present and future applications of lipases. Biotechnology and genetic engineering reviews, 3, 193–218.

    Article  CAS  Google Scholar 

  3. Jaeger, K. E., Dijkstra, B. W., & Reetz, M. T. (1999). Bacterial biocatalysts: Molecular biology, three-dimensional structures, and biotechnological applications of lipases. Annual Review of Microbiology, 53, 315–351.

    Article  CAS  Google Scholar 

  4. Saxena, R. K., Sheoran, A., Giri, B., & Davidson, W. S. (2003). Purification strategies for microbial lipases. Journal of Microbiological Methods, 52, 1–18.

    Article  CAS  Google Scholar 

  5. Singh, A. K., & Mukhopadhyay, M. (2012). Overview of fungal lipase: A review. Applied Biochemistry and Biotechnology, 166, 486–520.

    Article  CAS  Google Scholar 

  6. Ramnath, L., Sithole, B., & Govinden, R. (2017). Classification of lipolytic enzymes and their biotechnological applications in the pulping industry. Canadian Journal of Microbiology, 63, 179–192.

    Article  CAS  Google Scholar 

  7. Okal, E. J., Aslam, M. M., Karanja, J. K., & Nyimbo, W. J. (2020). Mini review: Advances in understanding regulation of cellulase enzyme in white-rot basidiomycetes. Microbial Pathogenesis, 147, 104410.

    Article  CAS  Google Scholar 

  8. Li, X., & Zheng, Y. (2020). Biotransformation of lignin: Mechanisms, applications and future work. Biotechnology Progress, 36, e2922.

  9. Sjostrom, E. (1993). In Wood Chemistry (Second Edition), (Sjostrom, E., ed.), Academic Press, San Diego, pp. 51–108.

  10. Gutiérrez, A., del Río, J. C., Martínez, M. J., & Martínez, A. T. (1999). Fungal degradation of lipophilic extractives in Eucalyptus globulus wood. Applied and Environmental Microbiology, 65, 1367–1371.

    Article  Google Scholar 

  11. Gutierrez, A., del Rio, J. C., & Martinez, A. T. (2009). Microbial and enzymatic control of pitch in the pulp and paper industry. Applied Microbiology and Biotechnology, 82, 1005–1018.

    Article  CAS  Google Scholar 

  12. Cohen, R., Persky, L., & Hadar, Y. (2002). Biotechnological applications and potential of wood-degrading mushrooms of the genus Pleurotus. Applied Microbiology and Biotechnology, 58, 582–594.

    Article  CAS  Google Scholar 

  13. Tellez-Tellez, M., & Diaz-Godinez, G. (2019). Omic tools to study enzyme production from fungi in the Pleurotus genus. BioResources, 14, 2420–2457.

    Article  CAS  Google Scholar 

  14. Skocaj, M., Gregori, A., Grundner, M., Sepcic, K., & Sezun, M. (2018). Hydrolytic and oxidative enzyme production through cultivation of Pleurotus ostreatus on pulp and paper industry wastes. Holzforschung, 72, 813–817.

    Article  CAS  Google Scholar 

  15. Piscitelli, A., Tarallo, V., Guarino, L., Sannia, G., Birolo, L., & Pezzella, C. (2017). New lipases by mining of Pleurotus ostreatus genome. PLoS ONE, 12, 15.

    Article  Google Scholar 

  16. Antošová, Z., & Sychrová, H. (2016). Yeast hosts for the production of recombinant laccases: A review. Molecular Biotechnology, 58, 93–116.

    Article  Google Scholar 

  17. Strinska, H. N., Petrov, D. N., Panajotova, H. N., Dobreva, V. T., Zhekova, B. J., & Dobrev, G. T. (2017). Isolation and purification of lipase from Rhizopus arrhizus by ultrafiltration and fractional precipitation. Bulgarian Chemical Communications, 49, 137–143.

    Google Scholar 

  18. Riley, R., Salamov, A. A., Brown, D. W., Nagy, L. G., Floudas, D., Held, B. W., Levasseur, A., Lombard, V., Morin, E., Otillar, R., Lindquist, E. A., Sun, H., LaButti, K. M., Schmutz, J., Jabbour, D., Luo, H., Baker, S. E., Pisabarro, A. G., Walton, J. D., … Grigoriev, I. V. (2014). Extensive sampling of basidiomycete genomes demonstrates inadequacy of the white-rot/brown-rot paradigm for wood decay fungi. Proc Natl Acad Sci U S A, 111, 9923–9928.

    Article  CAS  Google Scholar 

  19. Castanera, R., López-Varas, L., Borgognone, A., LaButti, K., Lapidus, A., Schmutz, J., Grimwood, J., Pérez, G., Pisabarro, A. G., Grigoriev, I. V., Stajich, J. E., & Ramírez, L. (2016). Transposable elements versus the fungal genome: Impact on whole-genome architecture and transcriptional profiles. PLoS Genetics, 12, e1006108

  20. Alfaro, M., Castanera, R., Lavín, J. L., Grigoriev, I. V., Oguiza, J. A., Ramírez, L., & Pisabarro, A. G. (2016). Comparative and transcriptional analysis of the predicted secretome in the lignocellulose-degrading basidiomycete fungus Pleurotus ostreatus. Environmental Microbiology, 18, 4710–4726.

    Article  CAS  Google Scholar 

  21. Verkasalo, E., Roitto, M., Möttönen, V., Tanner, J., Kumar, A., Kilpeläinen, P., Sikanen, L., & Ilvesniemi, H. (2022). Extractives of tree biomass of Scots pine (Pinus sylvestris L.) for biorefining in four climatic regions in Finland-lipophilic compounds, stilbenes, and lignans. Forests, 13, 779.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Applied Bioscience, Kanagawa Institute of Technology, 1030 Shimo-Ogino, Atsugi, Kanagawa, 243-0292, Japan

    Seiji Nakagame, Hu Minagawa & Nagi Motegi

Authors
  1. Seiji Nakagame
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hu Minagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nagi Motegi
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceptualization, S.N.; methodology, S.N., H.M, N.M.; validation, S.N., H.M, N.M.; investigation, S.N., H.M, N.M.; resources, S.N.; data curation, S.N., H.M, N.M.; writing-original draft preparation, S.N., H.M, N.M.; writing-review and editing, S.N.; visualization, S.N., H.M, N.M.; supervision, S.N.; project administration, S.N.; funding acquisition, S.N.

Corresponding author

Correspondence to Seiji Nakagame.

Ethics declarations

Ethics Approval

Not applicable.

Consent to Participate

Not applicable.

Consent for Publication

Not applicable.

Conflict of Interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nakagame, S., Minagawa, H. & Motegi, N. Purification and Characterization of Class III Lipase from a White-Rot Fungus Pleurotus ostreatus. Appl Biochem Biotechnol 195, 1085–1095 (2023). https://doi.org/10.1007/s12010-022-04211-0

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-022-04211-0

Keywords

Search

Navigation