Abstract
Cutinases are versatile carboxylic ester hydrolases with great potential in many biocatalytic processes, including biodiesel production. Genome sequence analysis of the model organism Aspergillus nidulans reveals four genes encoding putative cutinases. In this work, we purified and identified for the first time a cutinase (ANCUT2) produced by A. nidulans. ANCUT2 is a 29-kDa protein which consists of 255 amino acid residues. Comparison of the amino acid sequence of ANCUT2 with other microbial cutinase sequences revealed a high degree of homology with other fungal cutinases as well as new features, which include a serine-rich region and conserved cysteines. Cutinase production with different lipidic and carbon sources was also explored. Enzyme activity was induced by olive oil and some triacylglycerides and fatty acids, whereas it was repressed by glucose (1%) and other sugars. In some conditions, a 22-kDa post-translational processing product was also detected. The cutinase nature of the enzyme was confirmed after degradation of apple cutin.
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Abbreviations
- CEH:
-
Carboxylic ester hydrolase
- CEHM:
-
Carboxylic ester hydrolase medium
- h:
-
Hours
- RT:
-
Room temperature
- ATR:
-
Attenuated total reflectance
- DMSO:
-
Dimethyl sulfoxide
- NMWCO:
-
Nominal molecular weight cut-off
- TLC:
-
Thin-layer chromatography
- SDS:
-
Sodium dodecyl sulfate
- SDS–PAGE:
-
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
- PVDF:
-
Polyvinylidene fluoride
References
Webb, E. C. (1992). In International Union of Biochemistry and Molecular Biology, (Eds). San Diego: Academic. 0–12: 227164–227165.
Purdi, R. E., & Kolattukudy, P. E. (1975). Biochemistry, 14(13), 2824–2831.
Carvalho, C. M. L., Aires-Barros, M. R., & Cabral, J. M. (1999). Biotechnology and Bioengineering, 66(1), 17–34.
Panda, T., & Gowrishankar, B. S. (2005). Applied Microbiology and Biotechnology, 67, 160–169.
Sharma, R., Chisti, Y., & Banerjee, U. C. (2001). Biotechnology Advances, 19(8), 627–662.
Dutta, K., Sen, S., & Veeranki, V. D. (2009). Process Biochemistry, 44(2), 127–134.
Badenes, S. M., Lemos, F., & Cabral, J. M. (2010). Biotechnology Letters, 32, 399–403.
Badenes, S. M., Lemos, F., & Cabral, J. M. (2011). Biotechnology and Bioengineering, 108(6), 1279–1289.
Wang, X., Liu, X., Zhao, C., Ding, Y., & Xu, P. (2011). Bioresource Technology, 102(10), 6352–6355.
Kawasaki, L., Farrés, A., & Aguirre, J. (1995). Experimental Mycology, 19(1), 81–85.
García-Lepe, R., Nuero, O. M., Reyes, F., & Santamaría, F. (1997). Letters in Applied Microbiology, 25, 127–130.
Leger, R. J., St. Joshi, L., & Roberts, D. (1997). Microbiology, 143, 1983–1992.
Mayordomo, I., Randez-Gil, F., & Prieto, J. (2000). Journal of Agricultural and Food Chemistry, 48, 105–109.
Peña-Montes, C., Gonzalez, A., Castro-Ochoa, D., & Farrés, A. (2008). Applied Microbiology and Biotechnology, 78(4), 603–612.
Käfer, E. (1977). Advances in Genetics, 19, 33–131.
Walton, T. J., & Kolattukudy, P. E. (1972). Biochemistry, 11(10), 1885–1897.
Järvinen, R., Silvestre, A. J., Holopainen, U., Kaimainen, M., Nyyssölä, A., Gil, A. M., Pascoal-Neto, C., Lehtinen, P., Buchert, J., & Kallio, H. (2009). Journal of Agricultural and Food Chemistry, 57(19), 9016–9027.
Laemmli, U. K. (1970). Nature, 227, 680–685.
Bradford, M. M. (1976). Analytical Biochemistry, 72, 248–254.
Nielsen, H., Engelbrecht, J., Brunak, S., & von Heijne, G. (1997). Protein Engineering, 10, 1–6.
Blom, N., Gammeltoft, S., & Brunak, S. (1999). Journal of Molecular Biology, 294(5), 1351–1362.
Pearson, W. R. (1990). Methods in Enzymology, 183, 63–98.
Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., Lopez, R., Thomson, J. D., Gibson, T. J., & Higgings, D. G. (2007). Bioinformatics, 23, 2947–2948.
Saitou, N., & Nei, M. (1987). Molecular Biology and Evolution, 4, 406–425.
Felsenstein, J. (1985). Evolution, 39, 783–791.
Jones, D. T., Taylor, W. R., & Thornton, J. M. (1992). CABIOS, 8, 275–282.
Tamura, K., Dudley, J., Nei, M., & Kumar, S. (2007). Molecular Biology and Evolution, 24, 1596–1599.
Lin, T. S., & Kolattukudy, P. E. (1978). Journal of Bacteriology, 133(2), 942–951.
Pio, T. F., & Macedo, G. A. (2007). Enzyme and Microbial Technology, 41, 613–619.
Fett, W. F., Wijey, C., Moreau, R. A., & Osman, S. F. (1999). Journal of Applied Microbiology, 86, 561–568.
Fett, W. F., Wijey, C., Moreau, R. A., & Osman, S. F. (2000). Letters in Applied Microbiology, 31, 25–29.
Sarkar, S., Sreekanth, B., Kant, S., Banerjee, R., & Bhattacharyya, B. C. (1998). Bioprocess Engineering, 19, 29–32.
Henriette, C., Zinebi, S., Aumaitre, M. F., Petitdemange, E., & Petitdemange, H. (1993). Journal of Industrial Microbiology, 12, 129–135.
Martinez, P., Christen, P., & Farrés, A. (1993). Journal of Fermentation and Bioengineering, 76(2), 94–97.
Bauer, S., Vasu, P., Person, S., Mort, A. J., & Somerville, C. R. (2006). Proceedings of the National Academy of Sciences of the United States of America, 103(30), 11417–11422.
Lin, T., & Kolattukudy, P. E. (1980). European Journal of Biochemistry, 106, 341–351.
Wang, G. Y., Michailides, T. J., Hammock, B. D., Lee, Y. M., & Bostock, R. M. (2002). Fungal Genetics and Biology, 35(3), 261–276.
Kanazawa, T., Keeler, M., & Vartikovski, L. (1994). Cell Immunology, 156, 378–388.
Liu, Z., Gosser, Y., Baker, P. J., Ravee, Y., Lu, Z., Alemu, G., Li, H., Butterfoss, G. L., Kong, X. P., Gross, R., & Montclare, J. K. (2009). Journal of the American Chemical Society, 131(43), 15711–15716.
Galagan, J. E., et al. (2005). Nature, 438, 1105–1115.
Wang, G. Y., Michailides, T. J., Hammock, B. D., Lee, Y. M., & Bostock, R. M. (2000). Archives of Biochemistry and Biophysics, 382, 31–38.
Fan, C.-Y., & Köller, W. (1998). FEMS Microbiology Letters, 158, 33–38.
Pacchiano, R. A., Sohn, W., Chlanda, V. L., Garbow, J. R., & Stark, R. E. (1993). Journal of Agricultural and Food Chemistry, 41, 78–83.
Heredia-Guerrero, J. A., Heredia, A., García-Segura, R., & Benitez, J. J. (2009). Polymer, 50, 5633–5637.
Li, S., He, B., Bai, Z., & Ouyang, P. (2009). Journal of Molecular Catalysis B: Enzymatic, 56(2–3), 85–88.
Del Rio, J. C., & Hatcher, P. G. (1998). Organic Geochemistry, 29(5–7), 1441–1451.
Acknowledgments
Financial support for this project was obtained from PAPIIT–DGAPA–UNAM IN2148092. Denise Castro received a scholarship from CONACyT. We thank Dr. César Batista, from the Proteomics Unit, Biotechnology Institute, UNAM for LC–MS/MS Analysis and USAI Department, Chemistry Faculty, UNAM for infrared spectroscopy analysis. We appreciate the help of C. Warden for his assistance with the English revision of this manuscript.
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Castro-Ochoa, D., Peña-Montes, C., González-Canto, A. et al. ANCUT2, an Extracellular Cutinase from Aspergillus nidulans Induced by Olive Oil. Appl Biochem Biotechnol 166, 1275–1290 (2012). https://doi.org/10.1007/s12010-011-9513-7
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DOI: https://doi.org/10.1007/s12010-011-9513-7