DOI QR코드

DOI QR Code

Production of Alkaline Protease by Entrapped Bacillus licheniformis Cells in Repeated Batch Process

  • Received : 2011.05.19
  • Accepted : 2011.08.14
  • Published : 2011.12.28

Abstract

In this study, Bacillus licheniformis cells were immobilized by entrapment in calcium alginate beads and were used for production of alkaline protease by repeated batch process. In order to increase the stability of the beads, the immobilization procedure was optimized by statistical full factorial method, by which three factors including alginate type, calcium chloride concentration, and agitation speed were studied. Optimization of the enzyme production medium, by the Taguchi method, was also studied. The obtained results showed that optimization of the cell immobilization procedure and medium constituents significantly enhanced the production of alkaline protease. In comparison with the free-cell culture in pre-optimized medium, about 7.3-fold higher productivity was resulted after optimization of the overall procedure. Repeated batch mode of operation, using optimized conditions, resulted in continuous production of the alkaline protease for 13 batches in 19 days.

Keywords

References

  1. Abdel-Naby, M. A., A.-M. S. Ismail, S. A. Ahmed, and A. F. Abdel-Fattah. 1998. Production and immobilization of alkaline protease from Bacillus mycoides. Bioresource Technol. 64: 205-210. https://doi.org/10.1016/S0960-8524(97)00160-0
  2. Atalo, K. and B. A. Gashe. 1993. Protease production by a thermophilic Bacillus species (P-001A) which degrades various kinds of fibrous protein. Biotechnol. Lett. 15: 1151-1156. https://doi.org/10.1007/BF00131207
  3. Beshay, U. 2003. Production of alkaline protease by Teredinobacter turnirae cells immobilized in calcium alginate beads. Afr. J. Biotechnol. 2: 60-65.
  4. Chandran, S., S. Alagarsamy, G. Szakacs, and P. Ashok. 2005. Comparative evaluation of neutral protease production by Aspergillus oryzae in submerged and solid-state fermentation. Process Biochem. 40: 2689-2694. https://doi.org/10.1016/j.procbio.2004.12.001
  5. Calik, P., E. Bilir, G. Calik, and T. H. Ozdamar. 2002. Influence of pH conditions on metabolic regulations in serine alkaline protease production by Bacillus licheniformis. Enzyme Microb. Technol. 31: 685-697. https://doi.org/10.1016/S0141-0229(02)00162-X
  6. Chen, S. T., C. L. Kao, and K. T. Whang. 1995. Alkaline protease catalysis of a secondary amine to form a peptide bond. Int. J. Pept. Prot. Res. 46: 314-319.
  7. Germano, S., A. Pandey, C. A. Osaku, S. N. Rocha, and C. R. Soccol. 2003. Characterization and stability of proteases from Penicillium sp. produced by solid-state fermentation. Enzyme Microb. Technol. 32: 246-251. https://doi.org/10.1016/S0141-0229(02)00283-1
  8. Gessesse, A. 1997. The use of nug meal as low-cost substrate for the production of alkaline protease by the alkaliphilic Bacillus sp. AR-009 and some properties of the enzyme. Bioresource Technol. 62: 59-61. https://doi.org/10.1016/S0960-8524(97)00059-X
  9. Goksungur, V. and N. Zorlu. 2001. Production of ethanol from beet molasses by Ca-alginate immobilized yeast cells in a packed-bed bioreactor. Turk. J. Biol. 25: 265-275.
  10. Gombotz, W. R. and S. F. Wee. 1998. Protein release from alginate matrices. Adv. Drug Deliver. Rev. 31: 267-285. https://doi.org/10.1016/S0169-409X(97)00124-5
  11. Gupta, R., Q. K. Beg, S. Khan, and B. Chauhan. 2002. An overview on fermentation, downstream processing and properties of microbial alkaline proteases. Appl. Microbiol. Biotechnol. 60: 381-395. https://doi.org/10.1007/s00253-002-1142-1
  12. Haki, G. D. and S. K. Rakshit. 2003. Developments in industrially important thermostable enzymes: A review. Bioresource Technol. 89: 17-34. https://doi.org/10.1016/S0960-8524(03)00033-6
  13. Konsoula, Z. and M. Liakopoulou-Kyriakides. 2006. Thermostable $\alpha$-amylase production by Bacillus subtilis entrapped in calcium alginate gel beads. Enzyme Microb. Technol. 39: 690-696. https://doi.org/10.1016/j.enzmictec.2005.12.002
  14. Kumar, C. G. and H. Takagi.1999. Microbial alkaline proteases: From a bioindustrial viewpoint. Biotechnol. Adv. 17: 561-594. https://doi.org/10.1016/S0734-9750(99)00027-0
  15. Kunamneni, A., B. Jyothi, and P. Ellaiah. 2005. Production of alkaline protease with immobilized cells of Bacillus subtilis PE-11 in various matrices by entrapment technique. AAPS Pharmscitech. 6: 391-397. https://doi.org/10.1208/pt060348
  16. Kunitz, M. 1947. Protease assay. J. Gen. Physiol. 30: 291-310. https://doi.org/10.1085/jgp.30.4.291
  17. Lee, C. H., T. J. Kwon, S. M. Kang, H. H. Suh, G. S. Kwon, H. M. Oh, and B. D. Yoon. 1994. Production and characterization of an alkaline protease from an isolate, Xanthomonas sp. YL- 37. Korean J. Appl. Microbiol. Biotechnol. 22: 515-521.
  18. Mahmoud, D. A. and W. A. Helmy. 2009. Potential application of immobilization technology in enzyme and biomass production. J. Appl. Sci. Res. 5: 2466-2476.
  19. Malathi, S. and R. Chakraborty. 1991. Production of alkaline protease by a new Aspergillus flavus isolated under solidsubstrate fermentation conditions for use as a depilation agent. Appl. Environ. Microb. 27: 712-716.
  20. Mordocco, A. C., C. Kuek, and R. Jenkins. 1999. Continuous degradation of phenol at low concentration using immobilized Pseudomonas putida. Enzyme Microb. Technol. 25: 530-536. https://doi.org/10.1016/S0141-0229(99)00078-2
  21. Noori-Inanloo, D. and K. Rostami. 2000. Alkaline protease production by Bacillus licheniformis PTCC 1525 and characterization of some of the enzyme properties. The First National Biotechnology Congress of Islamic Republic of Iran, Iranian Research Organization for Science & Technology, Tehran, Iran.
  22. Oztop, H. N., A. Y. Oztop, E. Karadag, Y. Isikver, and D. Saraydin. 2003. Immobilization of Saccharomyces cerevisiae on to acrylamide sodium acrylate hydrogels for production of ethyl alcohol. Enzyme Microb. Technol. 32: 114-119. https://doi.org/10.1016/S0141-0229(02)00244-2
  23. Pandey, A., C. R. Soccol, J. A. Rodriguez-Leon, and P. Nigam. 2001. Solid State Fermentation in Biotechnology: Fundamentals and Applications. Academic Publishers, New Delhi.
  24. Potumarthi, R., Ch. Subhakar, and A. Jetty. 2007. Alkaline protease production by submerged fermentation in stirred tank reactor using Bacillus licheniformis NCIM-2042: Effect of aeration and agitation regimes. Biochem. Eng. J. 34: 185-192. https://doi.org/10.1016/j.bej.2006.12.003
  25. Potumarthi, R., Ch. Subhakar, A. Pavani, and A. Jetty. 2008. Evaluation of various parameters of calcium-alginate immobilization method for enhanced alkaline protease production by Bacillus licheniformis NCIM-2042 using statistical methods. Bioresource Technol. 99: 1776-1786. https://doi.org/10.1016/j.biortech.2007.03.041
  26. Quinn, G. P. and M. J. Keough. 2002. Experimental Design and Data Analysis for Biologists. Cambridge University Press, New York.
  27. Ramakrishna, S. V., R. Jamuna, and A. N. Emery. 1992. Production of ethanol by immobilized yeast. Appl. Biochem. Biotechnol. 37: 275-282. https://doi.org/10.1007/BF02788879
  28. Rossi-Alva, J. C. and M. H. M. Rocha-Leao. 2003. A strategic study using mutant-strain entrapment in calcium alginate for the production of Saccharomyces cerevisiae cells with high invertase activity. Biotechnol. Appl. Biochem. 38: 43-51. https://doi.org/10.1042/BA20020101
  29. Roy, R. K. 2001. Design of Experiments Using the Taguchi Approach: 16 Steps to Product and Process Improvement. Wiley-Interscience, New York.
  30. Vuillemard, J. C., J. Goulet, J. Amiot, M. A. Vijayalakshmi, and S. Terre. 1998. Continuous production of small peptides from milk proteins by extracellular proteases of free and immobilized Serratia marcescens cells. Enzyme Microb. Technol. 10: 2-8.

Cited by

  1. Monitoring the Ecology of Bacillus During Daqu Incubation, a Fermentation Starter, Using Culture-Dependent and Culture-Independent Methods vol.23, pp.5, 2013, https://doi.org/10.4014/jmb.1211.11065
  2. Enhanced production of protease by Pseudoalteromonas arctica PAMC 21717 via statistical optimization of mineral components and fed-batch fermentation vol.46, pp.4, 2011, https://doi.org/10.1080/10826068.2015.1031390
  3. Enhancement of the activity, stability and reusability of an extracellular protease from Pseudomonas fluorescens 07A via three different strategies of immobilization vol.37, pp.3, 2011, https://doi.org/10.1007/s43153-020-00059-1