Abstract
Factorial design and response surface techniques were used to optimize the culture medium for the production of inulinase by Kluyveromyces marxianus. Sucrose was used as the carbon source instead of inulin. Initially, a fractional factorial design (25–1) was used in order to determine the most relevant variables for enzyme production. Five parameters were studied (sucrose, peptone, yeast extract, pH, and K2HPO4), and all were shown to be significant. Sucrose concentration and pH had negative effects on inulinase production, whereas peptone, yeast extract, and K2HPO4 had positive ones. The pH was shown to be the most significant variable and should be preferentially maintained at 3.5. According to the results from the first factorial design, sucrose, peptone, and yeast extract concentrations were selected to be utilized in a full factorial design. The optimum conditions for a higher enzymatic activity were then determined: 14 g/L of sucrose, 10 g/L of yeast extract, 20 g/L of peptone, 1 g/L of K2HPO4. The enzymatic activity in the culture conditions was 127 U/mL, about six times higher than before the optimization.
Similar content being viewed by others
References
Vandamme, E. J. and Derycke, D. G. (1993), Adv. Appl. Microbiol. 29, 139–176.
Zittan, L. (1981), Starch 33, 373–377.
Gupta, A. K., Davinder, P. S., Kaur, N., and Singh, R. (1994), J. Chem. Technol. Biotechnol. 59, 377–385.
Kaur, N., Kaur, M., Gupta, A. K., and Singh, R. (1992), J. Chem. Technol. Biotechnol. 53, 279–284.
Poorna, V. and Kulkarni, P. R. (1995), Bioresour. Technol. 54, 315–320.
Nakamura, T., Shitara, A., Matsuda, S., Matsuo, T., Suiko, M., and Ohta, K. (1997), J. Ferment. Bioeng. 84, 313–318.
Parekh, S. and Margaritis, A. (1986), Agric. Biol. Chem. 50, 1085–1087.
Guerrero, A. C., Pena, I. G., Barzana, E., Garibay, M. G., and Ruiz, L. G. (1995), J. Ferment. Bioeng. 80, 159–163.
Selvakumar, P. and Pandey, A. (1999), Bioresour. Technol. 69, 123–127.
Rouwenhorst, R. J., Hensing, M., Verbakel, J., Scheffers, W. A., and Van Dijken, J. P. (1990), Appl. Environ. Microbiol. 56, 3337–3345.
Kim, D. H., Choi, Y. J., Song, S. K., and Yun, J. W. (1997), Biotechnol. Lett. 19, 369–371.
Box, G. E. P., Hunter, W. G., and Hunter, J. S. (1978), Statistics for Experimenters: An Introduction to Design, Data Analysis, and Model Building, John Wiley & Sons, New York.
Miller, G. L. (1959), Analyt. Chem. 31, 426–428.
Vandamme, E. J. and Derycke, D. G. (1983), Adv. Appl. Microbiol. 29, 139–176.
Pessoa, A., Jr., Hartmann, R., Vitolo, M., and Hustedt, H. (1996), J. Biotechnol. 51, 89–95.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kalil, S.J., Suzan, R., Maugeri, F. et al. Optimization of inulinase production by Kluyveromyces marxianus using factorial design. Appl Biochem Biotechnol 94, 257–264 (2001). https://doi.org/10.1385/ABAB:94:3:257
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1385/ABAB:94:3:257