Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Evolutionary Operation (EVOP) to Optimize Whey-Independent Serratiopeptidase Production from Serratia marcescens NRRL B-23112

  • Pansuriya, Ruchir C. (Food Engineering and Technology Department, Institute of Chemical Technology, University of Mumbai) ;
  • Singhal, Rekha S. (Food Engineering and Technology Department, Institute of Chemical Technology, University of Mumbai)
  • Received : 2009.11.23
  • Accepted : 2010.02.24
  • Published : 2010.05.28
Download PDF
⟨ Previous Next ⟩

Abstract

Serratiopeptidase (SRP), a 50 kDa metalloprotease produced from Serratia marcescens species, is a drug with potent anti-inflammatory property. In this study, a powerful statistical design, evolutionary operation (EVOP), was applied to optimize the media composition for SRP production in shake-flask culture of Serratia marcescens NRRL B-23112. Initially, factors such as inoculum size, initial pH, carbon source, and organic nitrogen source were optimized using one factor at a time. The most significant medium components affecting the production of SRP were identified as maltose, soybean meal, and $K_2HPO_4$. The SRP so produced was not found to be dependent on whey protein, but rather was notably induced by most of the organic nitrogen sources used in the study and free from other concomitant protease contaminant, as revealed by protease inhibition study. In addition, experiments were performed using different sets of EVOP design with each factor varied at three levels. The experimental data were analyzed with a standard set of statistical formula. The EVOP-optimized medium, with maltose 4.5%, soybean meal 6.5%, $K_2HPO_4$ 0.8%, and NaCl 0.5% (w/v), gave a SRP production of 7,333 EU/ml, which was 17-fold higher than the unoptimized media. The application of EVOP resulted in significant enhancement of SRP production.

Keywords

References

  1. Banerjee, R. 1990. Some studies on production purification and characterization of extracellular protease enzyme by Rhizopus oryzae. (RO, IIT KGP), Ph. D. Thesis, Indian Institute of Technology, Kharagpur, India.
  2. Banerjee, R. and B. C. Bhattacharyya. 1993. Evolutionary operation (EVOP) to optimize three-dimensional biological experiments. Biotechnol. Bioeng. 41: 67-71. https://doi.org/10.1002/bit.260410109
  3. Banerjee, R. and B. C. Bhattacharyya. 2003. Evolutionary operation as a tool of optimization for solid state fermentation. Biochem. Eng. J. 13: 149-155. https://doi.org/10.1016/S1369-703X(02)00127-4
  4. Baumann, U. 1994. Crystal structure of the 50 kDa metalloprotease from Serratia marcescens. J. Mol. Biol. 242: 244-251. https://doi.org/10.1006/jmbi.1994.1576
  5. Braun, V. and G. Schmitz. 1980. Excretion of a protease by Serratia marcescens. Arch. Microbiol. 124: 55-61. https://doi.org/10.1007/BF00407028
  6. Braunagel, S. C. and M. J. Benedik. 1990. The metalloprotease gene of Serratia marcescens strain SM6. Mol. Gen. Genet. 222: 446-451. https://doi.org/10.1007/BF00633854
  7. Bromke, B. J. and J. M. Hammel. 1979. Regulation of extracellular protease formation by Serratia marcescens. Can. J. Microbiol. 25: 47-52. https://doi.org/10.1139/m79-008
  8. Castaneda-Agullo, M. 1956. Studies on the biosynthesis of extracellular proteases by bacteria: I. Serratia marcescens, synthetic and gelatin media. J. Gen. Physiol. 39: 369-375 https://doi.org/10.1085/jgp.39.3.369
  9. Decedue, C. J., E. A. Broussard, A. D. Larson, and H. D. Braymer. 1979. Purification and characterization of the extracellular proteinase of Serratia marcescens. Biochem. Biophys. Acta 569: 293-301. https://doi.org/10.1016/0005-2744(79)90065-2
  10. Japanese Pharmacopoeia. 2006. Official monographs. 15th Ed. Part I. pp. 1735-1736.
  11. Klein, G. and W. Kullich. 2000. Short-term treatment of painful osteoarthritis of the knee with oral enzymes, a randomized, double-blind study versus diclofenac. Clin. Drug Invest. 19: 15-23. https://doi.org/10.2165/00044011-200019010-00003
  12. Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265-275.
  13. Mazzone, A., M. Catalani, and M. Costanzo. 1990. Evaluation of Serratia peptidase in acute or chronic inflammation of otorhinolaryngology pathology: A multicentre, double-blind, randomized trial versus placebo. J. Int. Med. Res. 18: 379-388. https://doi.org/10.1177/030006059001800506
  14. Miyata, K., K. Maejima, K. Tomoda, and M. Isono. 1970. Serratia protease Part I. Purification and general properties of the enzyme. Agric. Biol. Chem. 34: 310-318. https://doi.org/10.1271/bbb1961.34.310
  15. Miyata, K., K. Tomoda, and M. Isono. 1970. Serratia protease Part II. Substrate specificity of the enzyme. Agric. Biol. Chem. 34: 1457-1452. https://doi.org/10.1271/bbb1961.34.1457
  16. Miyata, K., K. Tomoda, and M. Isono. 1971. Serratia protease Part III. Characteristics of the enzyme as a metalloenzyme. Agric. Biol. Chem. 35: 460-467. https://doi.org/10.1271/bbb1961.35.460
  17. Miyazaki, H., N. Yanagida, S. Horinouchi, and T. Beppu. 1990. Specific excretion into the medium of a serine protease from Serratia marcescens. Agric. Biol. Chem. 54: 2763-2765. https://doi.org/10.1271/bbb1961.54.2763
  18. Nakahama, K., K. Yoshimura, R. Marumoto, M. Kikuchi, and I. S. Lee. 1986. Cloning and sequencing of Serratia protease gene. Nucleic Acids Res. 14: 5843-5855. https://doi.org/10.1093/nar/14.14.5843
  19. Romero, F., L. A. García, and M. Díaz. 1998. Protease production from whey at high concentrations by Serratia marcescens. Res. Environ. Biotechnol. 2: 93-115.
  20. Romero, F., L. A. García, J. A. Salas, M. Díaz, and L. M. Quirós. 2001. Production purification and partial characterization of two extracellular proteases from Serratia marcescens grown in whey. Process Biochem. 36: 501-515.
  21. Salamone, P. R. and R. J. Wodzinski. 1997. Production, purification and characterization of a 50-kDa extracellular metalloprotease from Serratia marcescens. Appl. Microbiol. Biotechnol. 48: 317-324. https://doi.org/10.1007/s002530051056
  22. Tunga, R., R. Banerjee, and B. C. Bhattacharyya. 1999. Optimization of n variable biological experiments by evolutionary operational-factorial technique. Biosci. Bioeng. 2: 224-230.
  23. Ustariz, J. F., A. G. Laca, and M. Diaz. 2004. Fermentation of individual proteins for protease production by Serratia marcescens. Biochem. Eng. J. 19: 147-153. https://doi.org/10.1016/j.bej.2003.12.009

Cited by

  1. Effects of Dissolved Oxygen and Agitation on Production of Serratiopeptidase by Serratia Marcescens NRRL B-23112 in Stirred Tank Bioreactor and its Kinetic Modeling vol.21, pp.4, 2010, https://doi.org/10.4014/jmb.1009.09031
  2. Enhanced Acetoin Production by Serratia marcescens H32 Using Statistical Optimization and a Two-stage Agitation Speed Control Strategy vol.17, pp.3, 2010, https://doi.org/10.1007/s12257-011-0587-4
  3. Statistical Optimization of Media Components for Production of Fibrinolytic Alkaline Metalloproteases from Xenorhabdus indica KB-3 vol.2014, pp.None, 2014, https://doi.org/10.1155/2014/293434
  4. Purification of serratiopeptidase from Serratia marcescens NRRL B 23112 using ultrasound assisted three phase partitioning vol.31, pp.None, 2010, https://doi.org/10.1016/j.ultsonch.2016.01.037
  5. Adaptation of CHO cells in serum‐free conditions for erythropoietin production: Application of EVOP technique for process optimization vol.63, pp.5, 2010, https://doi.org/10.1002/bab.1468
  6. Agro-industrial wastes utilization for the generation of fibrinolytic metalloprotease by Serratia marcescens RSPB11 vol.9, pp.None, 2010, https://doi.org/10.1016/j.bcab.2016.11.008
  7. Analytical techniques for serratiopeptidase: A review vol.7, pp.4, 2010, https://doi.org/10.1016/j.jpha.2017.03.005
  8. Investigation of the cell disruption methods for maximizing the extraction of arginase from mutant Bacillus licheniformis (M09) using statistical approach vol.35, pp.10, 2010, https://doi.org/10.1007/s11814-018-0107-8
  9. CpxR-Dependent Thermoregulation of Serratia marcescens PrtA Metalloprotease Expression and Its Contribution to Bacterial Biofilm Formation vol.200, pp.8, 2010, https://doi.org/10.1128/jb.00006-18
  10. Enhanced production of recombinant serratiopeptidase in Escherichia coli and its characterization as a potential biosimilar to native biotherapeutic counterpart vol.18, pp.None, 2010, https://doi.org/10.1186/s12934-019-1267-x