Abstract
A statistical approach, response surface methodology (RSM), was used to study the production of extracellular protease fromBacillus sp., which has properties of immense industrial importance. The most influential parameters for protease production obtained through the method of testing the parameters one at a time were starch, soybean meal, CaCl2, agitation rate, and inoculum density. This method resulted in the production of 2543 U/mL of protease in 48 h fromBacillus sp. Based on these results, face-centered central composite design falling under RSM was employed to further enhance protease activity. The interactive effect of the most influential parameters resulted in a 1.50-fold increase in protease production, yielding 3746 U/mL in 48 h. Analysis of variance showed the adequacy of the model and verification experiments confirmed its validity. On subsequent scale-up in a 30-L bioreactor using conditions optimized through RSM, 3978 U/mL of protease was produced in 18 h. This clearly indicated that the model remained valid even on a large scale. RSM is a quick process for optimization of a large number of variables and provides profound insight into the interactive effect of various parameters involved in protease production.
Similar content being viewed by others
References
Godfrey, T. and West, S. (1996), inIndustrial Enzymology, 2nd Edition (Godfrey, T. and West, S., eds.) Macmillan, UK: pp. 1–8.
Rao, M. B., Tanksale, A. M., Ghatke, M. S., and Deshpande, V. V. (1998), inMicrobiology Molecular Biology Reviews,62, 597–635.
Takac, S., Elmas, S., Çalik, P., and Özdamar, T. H. (2000),J. Chem. Technol. Biotechnol. 75, 491–499.
Kumar, C. G. and Takagi, H. (1998),Biotechnol. Adv. 17, 561–594.
Adinarayana, K. and Ellaiah, P. (2002),J. Pharm. Sci. 5, 272–278.
Masse, F. W. J. L. and Tilburg, R. V. (1983),J. Am. Oil Chem. Soc. 60, 1672–1675.
Prakasham, R. S., Rao, C. S., Rao, R. S., and Sharma, P. N. (2005),Biotechnol. Prog. 21(5), 1380–1388.
Joo, H. S., Kumar, C. G., Park, G. C., Kim, K. M., Paik, S. R., and Chang, C. S. (2002),Process Biochem. 38, 155–159.
Manachini, P. L. and Fortina, M. G. (1998),Biotechnol. Lett. 20, 565–568.
Jacobs, M. F. (1995),Gene 152, 67–74.
Yang, J. K., Shih, I. L., Tzeng, Y. M., and Wang, S. L. (2000),Enzyme Microb. Technol. 26, 406–413.
Ito, S., Kobayashi, T., Ara, K., Ozaki, K., Kawai, S., and Hatada, Y. (1998),Extremophile 2, 185–190.
Beg, Q. K., Sahai, V., and Gupta, R. (2003),Process Biochem. 38, 1–7.
Kaur, S., Vohra, R. M., Kapoor, M., Beg, Q. K., and Hoondal, G. S. (2001),J. Gen. Microbiol. 128, 845–851.
McKeller, R. C. and Cholete, H. (1984),Appl. Microbiol. Biotechnol. 47, 1224–1227.
Puri, S., Beg, Q. K., and Gupta, R. (2002),Curr. Microbiol. 44, 286–290.
Nehete, P. N., Shah, V. D., and Kothari, R. M. (1985),Biotechnol. Lett. 7, 413–418.
Oberoi, R., Beg, Q. K., Puri, S., Saxena, R. K., and Gupta, R. (2001),World J. Microbiol. Biotechnol. 60, 381–395.
Razak, N. A., Samad, M. Y. A., Basri, M., Yunus, W. M. Z. W., Ampon, K., and Salleh, A. B. (1994),World J. Microbiol. Biotechnol. 10, 260–263.
Saxena, S. and Saxena, R. K. (2004),Biotechnol. Appl. Biochem. 39, 99–106.
Vohra, A. and Satanarayana, T. (2002),Process Biochem. 37, 999–1004.
Kalil, S. J., Maugeri, F., and Rodrigues, M. I. (2000),Process Biochem. 35, 539–550.
Haaland, P. D. (1989), inExperimental Design in Biotechnology, (Halland P.D., ed) Marcel Dekker, New York, pp. 1–18.
Krishna, S. H., Manohar, B., Divakar, S., Prapulla, S. G., and Karanth, N. G. (2000),Enzyme Microb. Technol. 26, 131–136.
Isar, J., Agarwal, L., Saran, S., and Saxena, R. K. (2006),Bioresour. Technol., 97, 1443–1448.
Meyers, S. P. and Ahearn, D. G. (1977),Mycologia 69, 646–651.
Chauhan, B. and Gupta, R. (2004),Process Biochem. 39, 2115–2122.
Burkert, J. F., Maureri, M. F., and Rodrigues, M. I. (2004),Bioresour. Technol. 91, 77–84.
Moon, S. H. and Parulekar, S. J. (1991),Biotechnol. Bioeng. 37, 467–483.
Chu, I. M., Lee, C., and Li, T. S. (1992),Enzyme Microb. Technol. 14, 755–761.
Gupta, R., Beg, Q. K., Khan, S., and Chauhan, B. (2002),Appl. Microbiol. Biotechnol. 60, 381–395.
Jang, J. W., Jun, H. K., Kim, E. K., Jang, W. H., Kang, J. H., and Yoo, O. J. (2001),Biotechnol. Appl. Biochem. 34, 81–84.
Hameed, A., Keshavarz, T., and Evabs, C. S. (1999),J. Chem. Technol. Biotechnol. 74, 5–8.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Saran, S., Isar, J. & Saxena, R.K. Statistical optimization of conditions for protease production fromBacillus sp. and its scale-up in a bioreactor. Appl Biochem Biotechnol 141, 229–239 (2007). https://doi.org/10.1007/BF02729064
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02729064