Abstract
Among physical and nutritional parameters optimized by “one variable at a time” approach, four cultural variables (sucrose, MgSO4 .7H2O, inoculum size, and incubation period) significantly affected glucoamylase production. These variables were, therefore, selected for optimization using response surface methodology. The p-values of the coefficients for linear effect of sucrose and inoculum size were less than 0.0001, suggesting them to be the key experimental variables in glucoamylase production. The enzyme production (34 U/ml) attained under optimized conditions (sucrose, 2%; MgSO4 .7H2O, 0.13%; yeast extract, 0.1%; inoculum size, 5 × 106 spores per 50 ml production medium; incubation time, 48 h; temperature, 40°C; and pH 7.0) was comparable with the value predicted by polynomial model (34.2 U/ml). An over all 3.1-fold higher enzyme titers were attained due to response surface optimization. The experimental model was validated by carrying out glucoamylase production in shake flasks of increasing capacity (0.25–2.0 l) and 22-l laboratory bioreactors (stirred tank and airlift), where the enzyme production was sustainable. Furthermore, the fermentation time was reduced from 48 h in shake flasks to 32 h in bioreactors.
Similar content being viewed by others
References
Angenent, L. T., Karim, K., Al-Dahhan, M. H., Wrenn, B. A., & Domiguez-Espinosa, R. (2004). Trends in Biotechnology, 22, 477–485.
Crabb, W. D., & Shetty, J. K. (1999). Current Opinion in Microbiology, 2, 252–256.
Archer, D. B. (2000). Current Opinion in Biotechnology, 11, 478–483.
Maarel, M. J. E. C., Veen, B. V. D., Uitdehaag, J. C. M., Leemhuis H., & Dijkhuijen, L. (2002). Journal of Biotechnology, 94, 137–155.
Reilly, P. J. (1999). Starch/Starke, 51, 269–274.
Pandey, A. (1995). Starch/Starke, 47, 439–445.
Vieille, C., & Zeikus, G. J. (2001). Microbiology and Molecular Biology Reveiws, 65, 1–43.
Vihinen, M., & Mantsala, P. (1989). CRC Critical Reviews in Biochemisty and Molecular Biology, 24, 329–419.
Thorsen, T. S., Johnsen, A. H., Josefsen, K., & Jensen, B. (2006). Biochimica et Biophysica Acta, 1764, 671–676.
Pham, P. L., Taillandier, P., Delmas, M., & Strehaiano, P. (1998). World Journal of Microbiolology and Biotechnology, 14, 185–190.
Cochran, W. G, & Cox, D. W. (1968). Experimental Designs. New York: John-Wiley and Sons. Inc.
Myers, R. H., & Montgomery, D. C. (1995). Response Surface Methodology: Process and Product Optimization Using Designed Experiments. New York: J. Wiley & Sons.
Gu, X.-B., Zheng, Z.-M., Yu, H.-Q., Wang, J., Liang, F.-L., & Liu, R.-L. (2005). Process Biochemistry, 40, 3196–3201.
Kumar, S., & Satyanarayana, T. (2001). World Journal of Microbiology and Biotechnology, 17, 83–87.
Uma Maheswar Rao, J. L., & Satyanarayana, T. (2006). Bioresource Technology, 98, 345–352.
Satyanarayana, T., Noorwez, S. M., Kumar, S., Rao, J. L. U. M., Ezhilvannan, M., & Kaur, P. (2004). Biochemical Society Transactions, 32, 276–279.
Subrahmanyam, A., Mehrotra, B. S., & Thirumalachar, M. J. (1977). Georgia Journal of Science, 35, 106.
Emerson, R. (1941). Lloydia, 4, 77–144.
Miller, G. L. (1959). Analytical Chemistry, 31, 426–428.
Lowry, O. W., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Journal of Biology and Chemistry, 193, 265–275.
Kaur, P., & Satyanarayana, T. (2004). World Journal of Microbiology and Biotechnology, 20, 419–425.
Kumar, S., & Satyanarayana, T. (2003). Biotechnology Progress, 19, 936–944.
Moreira, F. G., de Lima, F. A., Pedrinho, S. R. F., Lenartovicz, V., de Souza, C. G. M., & Peralta, R. M. (1999). Revista de Microbiologia, 30, 157–162.
Fattah, Y. R. A. (2002). Biotechnology Letters, 24, 1222–2002.
Francis, F., Sabu, A., Nampoothiri, K. M., Ramachandran, S., Ghosh, S., Szakacs, G., et al. (2003). Biochemical Engineering Journal, 15, 107–115.
Vaidya, R., Vyas, P., & Chhatpar, H. S. (2003). Enzyme and Microbial Technology, 33, 92–96.
Babu, K. R., & Satyanarayana, T. (1993). Enzyme and Microbial Technology, 15, 1066–1069.
Humphrey, A. (1998). Biotechnology Progress, 14, 3–7.
Sharma, D. C., & Satyanarayana, T. (2006). Bioresource Technology, 97, 727–733.
Rao, Y. K., Lu, S.-C., Liu, B.-L., & Tzeng, Y.-M. (2006). Biochemical Engineering Journal, 28, 57–66.
Morkeberg, R., Carlsen, M., & Nielsen, J. (1995). Microbiology (United Kingdom), 141, 1449–1454.
Narang, S., & Satyanarayana, T. (2001). Letters in Applied Micribiology, 32, 31–35.
Acknowledgements
We are grateful to the University Grants Commission, Government of India for providing financial assistance. SK and PK thank the Council of Scientific and Industrial Research, Government of India for awarding fellowship during the course of this investigation.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kumar, S., Kumar, P. & Satyanarayana, T. Production of Raw Starch-Saccharifying Thermostable and Neutral Glucoamylase by the Thermophilic Mold Thermomucor indicae-seudaticae in Submerged Fermentation. Appl Biochem Biotechnol 142, 221–230 (2007). https://doi.org/10.1007/s12010-007-0011-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-007-0011-x