Evolutionary operation (EVOP) to optimize whey independent serratiopeptidase production from Serratia marcescens NRRL B-23112.

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. Most significant medium components affecting the production of SRP were identified as maltose, soybean meal and KHPO. The SRP so produced was not found to be dependent on whey protein, rather notably induced by most of the organic nitrogen sources used in the study and free from other concomitant protease contaminant revealed by protease inhibition study. Further, experiments were performed using different sets of EVOP design with each factor varied at three levels. The experimental data were analyzed with standard set of statistical formula. The EVOP optimized medium, maltose 4.5%, soybean meal 6.5%, KHPO 0.8% and NaCl 0.5% w/v gave 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.

[1]  M. Díaz,et al.  Fermentation of individual proteins for protease production by Serratia marcescens , 2004 .

[2]  Rintu Banerjee,et al.  Evolutionary operation as a tool of optimization for solid state fermentation , 2003 .

[3]  P. Salamone,et al.  Production, purification and characterization of a 50-kDa extracellular metalloprotease from Serratia marcescens , 1997, Applied Microbiology and Biotechnology.

[4]  U. Baumann,et al.  Crystal structure of the 50 kDa metallo protease from Serratia marcescens. , 1994, Journal of molecular biology.

[5]  B. C. Bhattacharyya,et al.  Evolutionary operation (EVOP) to optimize three‐dimensional biological experiments , 1993, Biotechnology and bioengineering.

[6]  S. Horinouchi,et al.  Specific Excretion into the Medium of a Serine Protease from Serratia marcescens , 1990 .

[7]  A. Mandoli,et al.  Evaluation of Serratia Peptidase in Acute or Chronic Inflammation of Otorhinolaryngology Pathology: A Multicentre, Double-Blind, Randomized Trial versus Placebo , 1990, The Journal of international medical research.

[8]  S. C. Braunagel,et al.  The metalloprotease gene ofSerratia marcescens strain SM6 , 1990, Molecular and General Genetics MGG.

[9]  H. Matsubara,et al.  Cloning and sequencing of Serratia protease gene. , 1986, Nucleic acids research.

[10]  A. D. Larson,et al.  Purification and characterization of the extracellular proteinase of Serratia marcescens. , 1979, Biochimica et biophysica acta.

[11]  K. Tomoda,et al.  Serratia Protease: Part III. Characteristics of the Enzyme as Metalloenzyme , 1971 .

[12]  K. Tomoda,et al.  Serratia Protease: Part II. Substrate Specificity of the Enzyme , 1970 .

[13]  K. Tomoda,et al.  Serratia Protease:Part I. Purification and General Properties of the Enzyme , 1970 .

[14]  M. Castañeda-Agulló.  STUDIES ON THE BIOSYNTHESIS OF EXTRACELLULAR PROTEASES BY BACTERIA , 1956, The Journal of general physiology.

[15]  Oliver H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[16]  V. Braun,et al.  Excretion of a protease by Serratia marcescens , 2004, Archives of Microbiology.

[17]  M. CastaiDa-Agull.  STUDIES ON THE BIOSYNTHESIS OF EXTRACELLULAR PROTEASES BY BACTERIA I. S E ~ T ~ MARCESCENS. SYNTHETIC AND GELATIN MEDIA* , 2003 .

[18]  Luis A. García,et al.  Production, purification and partial characterization of two extracellular proteases from Serratia marcescens grown in whey , 2001 .

[19]  G. Klein,et al.  Short-Term Treatment of Painful Osteoarthritis of the Knee with Oral Enzymes , 2000 .

[20]  B. C. Bhattacharyya,et al.  Optimization of n variable biological experiments by evolutionary operation-factorial design technique. , 1999, Journal of bioscience and bioengineering.

[21]  M. Díaz,et al.  Protease production from whey at high concentrations by Serratia marcescens , 1998 .

[22]  J. Hammel,et al.  Regulation of extracellular protease formation by Serratia marcescens. , 1979, Canadian journal of microbiology.