Abstract
The aim of this experimental study was to isolate and partially purify protease enzyme from Bacillus cereus and Bacillus subtilis. Protease enzyme is obtained by inducing spore genesis of bacteria from Bacillus species in suitable nutrient plates. The partial purification was realized by applying, respectively, ammonium sulfate precipitation, dialysis, and DEAE-cellulose ion-exchange chromatography to the supernatant that was produced later. Optimum pH, optimum temperature, pH stability, and temperature stability were determined, as well as the effects of pH, temperature, substrate concentration, reaction time, and inhibitors and activators on enzyme activity. In addition, the molecular mass of the obtained enzyme was investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The specific activity of partially purified enzyme from B. subtilis was determined to be 84 U/mg. The final enzyme preparation was eight-fold more pure than the crude homogenate. The molecular mass of the partially purified enzyme was found to be 45 kDa by using SDS-PAGE. The protease enzyme that was partially purified from B. cereus was purified 1.2-fold after ammonium sulfate precipitation. The molecular mass of the partially purified enzyme was determined to be 37 kDa by using SDS-PAGE.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Kole, M. M., Draper, L, and Gerson, D. F. (1988), J. Chem. Technol. Biotechnol. 41, 197–206.
Jadwiga, K. and Sierecka, K. (1998), Int. J. Biochem. Cell Biol. 30, 579–595.
Banerjee, C. U., Sani, R. K., Azmi, W., and Soni, R. (1999), Process Biochem. 35, 213–219.
Romero, F. J., Garcia, L. A., Salas, J. A., Diaz, M., and Quiros, L. M. (2001), Process Biochem. 36, 507–515.
Singh, J., Ahmed, S. (2001), Process Biochem. 36, 781–785.
Matta, H. and Punj, V. (1998), Int. J. Food Microbiol. 42, 139–145.
Gerhartz, W. (1990), Enzyme in Industry, VCH, Weinheim, Germany.
James, H. (2000), Biotechnol. Appl. Biochem. 31, 85–89.
Kumar, C. G. (2002), Lett. Appl. Microbiol. 34, 13–17.
Kumar, C. G. and Takagi, H. (1999), Biotechnol. Adv. 17, 561–594.
Price, N. C. and Stevens, L. (1982), Fundamentals of Enzymology, Oxford University Press, Oxford, England.
Trevor, P. (1987), Enzymes, Science and Technology Press, Ankara, Turkey.
Ozikara, E. (1999), Biochemistry, Ankara University Press, Ankara, Turkey.
Jen-Kuo, Y. (2000), Enzyme Microb. Technol. 26, 406–413.
Dhandapani, R. and Vijayaragavan, R. (1994), World J. Microbiol. Biotechnol. 10, 33–35.
Stoscheck, C. M. (1990), Methods in Enzymol. 182, 50–69.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this chapter
Cite this chapter
Orhan, E., Omay, D., Güveniur, Y. (2005). Partial Purification and Characterization of Protease Enzyme from Bacillus subtilis and Bacillus cereus . In: Davison, B.H., Evans, B.R., Finkelstein, M., McMillan, J.D. (eds) Twenty-Sixth Symposium on Biotechnology for Fuels and Chemicals. ABAB Symposium. Humana Press. https://doi.org/10.1007/978-1-59259-991-2_16
Download citation
DOI: https://doi.org/10.1007/978-1-59259-991-2_16
Publisher Name: Humana Press
Print ISBN: 978-1-58829-697-9
Online ISBN: 978-1-59259-991-2
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)