Development of Medium and Kinetic Modeling for Enhanced Production of Cutinase from Pseudomonas cepacia NRRL B-2320

Kasturi Dutta, Veeranki Venkata Dasu, Krishnamoorthy Hegde

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Biochemical Engineering Laboratory, Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, India.

DOI: 10.4236/aim.2013.36064                

Abstract

The development of medium for the production of cutinase from Pseudomonas cepacia NRRL B 2320 was carried out using Plackett-Burman experimental design followed by central composite design. The medium components were screened by Plackett-Burman experimental design which suggested that cutin, peptone, KCl and MgSO₄·7H₂O have influenced the cutinase production significantly with very high confidence levels. The concentration levels of these four components were optimized using 2⁴ full factorial central composite design. An optimum combination of 10.06 g·L^-1 of cutin, 17.77 g·L^-1 of peptone, 0.635 g·L^-1 of KCl and 5.455 g·L^-1 of MgSO₄·7H₂O in the medium gave a maximum cutinase activity of 336 U·mL^-1. An overall 2 fold increase in the production of cutinase was observed in the optimized medium. Growth and production of cutinase from P. cepacia NRRL B 2320 have been studied in shake flask and batch bioreactor. Time course of cell growth and enzyme production was fitted to the existing kinetic models reported in the literature to estimate the biokinetic parameters. These models suggested that the production of cutinase is growth associated in shake flask and it is a mixed growth type in a batch bioreactor.

Keywords

Cutinase; Kinetics; Bioprocessing; Optimization; Parameter Identification

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	K. Dutta, S. Sen and V. D. Veeranki, “Production, Characterization and Applications of Microbial Cutinases,” Process Biochemistry, Vol. 44, No. 2, 2009, pp. 127-134. http://dx.doi.org/10.1016/j.procbio.2008.09.008
[2]	C. M. L. Carvalho, M. R. Aires-Barros and J. M. S. Cabral, “Cutinase: From Molecular Level to Bioprocess Development,” Biotechnology and Bioengineering, Vol. 66, No. 1, 1999, pp. 17-34. http://dx.doi.org/10.1002/(SICI)1097-0290(1999)66:1<17::AID-BIT2>3.0.CO;2-F
[3]	R. E. Purdy and P. E. Kolattukudy, “Hydrolysis of Plant Cuticle by Plant Pathogens. Purification, Amino-Acid Composition, and Molecular Weight of 2 Isoenzymes of Cutinase and a Nonspecific Esterase from Fusarium solani f. pisi,” Biochemistry, Vol. 14, No. 13, 1975, pp. 2824-2831. http://dx.doi.org/10.1021/bi00684a006
[4]	C. G. Dekoster, W. Heerma, H. A. M. Pepermans, A. Groenewegen, H. Peters and J. Haverkamp, “Tandem Mass Spectrometry and Nuclear Magnetic Resonance Spectroscopy Studies of Candida bombicola Sophorolipids and Product Formed on Hydrolysis by Cutinase,” Analytical Biochemistry, Vol. 230, No. 1, 1995, pp. 135-148. http://dx.doi.org/10.1006/abio.1995.1448
[5]	C. M. L. Carvalho, J. M. S. Cabral and M. R. Aires-Barros, “Kinetics and Modelling of Transesterification Reactions Catalysed by Cutinase in AOT Reversed Micelles,” Journal of Molecular Catalysis B: Enzymatic, Vol. 5, No. 1-4, 1998, pp. 361-365. http://dx.doi.org/10.1016/S1381-1177(98)00098-8
[6]	E. P. Melo, R. P. Baptista and J. M. S. Cabral, “Improving Cutinase Stability in Aqueous Solution and in Reverse Micelles by Media Engineering,” Journal of Molecular Catalysis B: Enzymatic, Vol. 22, No. 5-6, 2003, pp. 299-306. http://dx.doi.org/10.1016/S1381-1177(03)00044-4
[7]	V. V. Dasu and T. Panda, “Optimization of Microbiological Parameters for Enhanced Griseofulvin Production Using Response Surface Methodology,” Bioprocess Engineering, Vol. 22, No. 1, 2000, pp. 45-49. http://dx.doi.org/10.1007/PL00009099
[8]	S. Kumar, K. Pakshirajan and V. V. Dasu, “Development of Medium for Enhanced Production of Glutaminase-Free l-Asparaginase from Pectobacterium carotovorum MTCC 1428,” Applied Microbiology and Biotechnology, Vol. 84, No. 3, 2009, pp. 477-486. http://dx.doi.org/10.1007/s00253-009-1973-0
[9]	D. C. Sharma and T. Satyanarayana, “A Marked Enhancement in the Production of a Highly Alkaline and Thermostable Pectinase by Bacillus pumilus dcsr1 in Submerged Fermentation by Using Statistical Methods,” Bioresource Technology, Vol. 97, No. 5, 2006, pp. 727-733. http://dx.doi.org/10.1016/j.biortech.2005.04.012
[10]	T. J. Walton and P. E. Kolattukudy, “Determination of Structures of Cutin Monomers by a Novel Depolymerization Procedure and Combined Gas Chromatography and Mass Spectrometry,” Biochemistry, Vol. 11, No. 10, 1972, pp. 1885-1897. http://dx.doi.org/10.1021/bi00760a025
[11]	M. M. A. Elsawah, A. A. Sherief and S. M. Bayoumy, “Enzymatic Properties of Lipase and Characteristics Production by Lactobacillus delbrueckii Subsp. Bulgaricus,” Antonie van Leeuwenhoek, Vol. 67, No. 4, 1995, pp. 357-362. http://dx.doi.org/10.1007/BF00872935
[12]	Degani, Ofir, H. Salman, S. Gepstein, G. Carlos and Dosoretz. “Synthesis and Characterization of a New Cutinase Substrate, 4-Nitrophenyl (16-Methyl sulfone ester) Hexadecanoate,” Journal of Biotechnology, Vol. 121, No. 3, 2006, pp. 346-350. http://dx.doi.org/10.1016/j.jbiotec.2005.08.011
[13]	R. Agarwal, B. Mahanty and V. V. Dasu, “Modeling Growth of Cellulomonas cellulans NRRL B 4567 under Substrate Inhibition during Cellulase Production,” Chemical and Biochemical Engineering Quarterly, Vol. 23, No. 2, 2009, pp. 213-218.
[14]	R. L. Plackett and J. P. Burman, “The Design of Optimal Multifactorial Experiments,” Biometrika, Vol. 33, No. 4, 1946, pp. 305-325. http://dx.doi.org/10.1093/biomet/33.4.305
[15]	A. I. Khuri and J. A. Cornell, “Response Surfaces: Design and Analysis,” Marcel Decker Inc., New York, 1987.
[16]	P. W. Araujo and R. G. Brereton, “Experimental Design .2. Optimization,” TrAC. Trends in Analytical Chemistry, Vol. 15, No. 2, 1996, pp. 63-70.
[17]	P. Mercier, L. Yerushalmi, D. Rouleau and D. Dochain, “Kinetics of Lactic-Acid Fermentation on Glucose and Corn by Lactobacillus amylophilus,” Journal of Chemical Technology and Biotechnology, Vol. 55, No. 2, 1992, pp. 111-121. http://dx.doi.org/10.1002/jctb.280550204
[18]	R. Luedeking and E. L. Piret, “A Kinetic Study of the Lactic Acid Fermentation. Batch Process at Controlled pH,” Journal of Biochemistry, Vol. 1, No. 4, 1959, pp. 393-412.
[19]	J. Sebastian, A. K. Chandra and P. E. Kolattukudy, “Discovery of a Cutinase-Producing Pseudomonas sp. Cohabiting with an Apparently Nitrogen-Fixing Corynebacterium sp. in the Phyllosphere,” Journal of Bacteriology, Vol. 169, No. 1, 1987, pp. 131-136.
[20]	T. F. Pio and G. A. Macedo, “Optimizing the Production of Cutinase by Fusarium oxysporum Using Response Surface Methodology,” Enzyme Microbial Technology, Vol. 41, No. 5, 2007, pp. 613-619. http://dx.doi.org/10.1016/j.enzmictec.2007.05.008
[21]	F. J. Rispoli, and V. Shah, “Mixture Design as a First Step for Optimization of Fermentation Medium for Cutinase Production from Colletotrichum lindemuthianum,” Journal of Industrial Microbiology and Biotechnology, Vol. 34, No. 5, 2007, pp. 349-355. http://dx.doi.org/10.1007/s10295-007-0203-y
[22]	A. Rajendran, V. Selvaraj and V. Thangavelu, “Statistical Optmization and Kinetic Modeling of Xylanase Production by Arthrobacter sp.,” Asia-Pacific Journal of Chemical Engineering, Vol. 3, No. 3, 2008, pp. 347-353. http://dx.doi.org/10.1002/apj.138
[23]	A. Rajendran and V. Thangavelu, “Statistical Experimental Design for Evaluation of Medium Components for Lipase Production by Rhizopus arrhizus MTCC 2233,” LWT—Food Science and Technology, Vol. 42, No. 5, 2009, pp. 985-992.