Abstract
The microbial fermentation of Penicillium brevicompactum produces secondary metabolite mycophenolic acid (MPA), which exhibits antifungal, antiviral, antibacterial, and antitumor activity. It is also a potent, selective, non-competitive, and reversible inhibitor of the human inosine monophosphate dehydrogenase (IMPDH). This study is an attempt to optimize the MPA production through a fermentation process using Penicillium brevicompactum and its further purification process optimization. In the batch fermentation process, the maximum concentration of MPA (1.84 g/L) was attained in a 3.7 L stirred tank reactor. Response surface methodology (RSM) using central composite design (CCD) was employed as a statistical tool to investigate the effect of pH, the volume of eluent and flow rate of the mobile phase on MPA purification process. Under optimum conditions, the experimental yield was observed to be 84.12%, which matched well with the predictive yield of 84.42%. High-performance liquid chromatography (HPLC) and Fourier-transform infrared spectroscopy (FTIR) analysis of the fermented product was carried out to confirm the presence of mycophenolic acid. The MPA purification was done by using column chromatography technique. The purification of broth involved mycophenolic acid extraction by selecting different solvents on the basis of polarity and the extraction efficiency of solvent. Various solid support materials were used for MPA purification in column chromatography. The MPA recovery through alumina column was observed to be 84.12% under the optimum conditions, which was maximum elution as compared with other support materials. The optimized purification process yielded pure MPA crystals.
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References
Bentley, R. (2000). Mycophenolic acid: a one hundred year odyssey from antibiotic to immunosuppressant. Chemical Reviews, 100(10), 3801–3826.
Sydenstricker, V. P. (1958). The history of pellagra, its recognition as a disorder of nutrition and its conquest. The American Journal of Clinical Nutrition, 6(4), 409–414.
Ismaiel, A. A., Ahmed, A. S., & El-Sayed, E.-S. R. (2014). Optimization of submerged fermentation conditions for immunosuppressant mycophenolic acid production by Penicillium roqueforti isolated from blue-molded cheeses: Enhanced production by ultraviolet and gamma irradiation. World Journal of Microbiology and Biotechnology, 30(10), 2625–2638.
Sadhukhan, A., Murthy, M. R., Kumar, R. A., Mohan, E., Vandana, G., Bhar, C., & Rao, K. V. (1999). Optimization of mycophenolic acid production in solid state fermentation using response surface methodology. Journal of Industrial Microbiology & Biotechnology, 22, 33–38.
Puel, O., Tadrist, S., Galtier, P., Oswald, I. P., & Delaforge, M. (2005). Byssochlamys nivea as a source of mycophenolic acid. Applied and Environmental Microbiology, 71(1), 550–553.
Ardestani, F., & Fatemi, S. S.-a., Yakhchali, B., Hosseyni, S. M. and Najafpour, G. (2010). Evaluation of mycophenolic acid production by Penicillium brevicompactum MUCL 19011 in batch and continuous submerged cultures. Biochemical Engineering Journal, 50, 99–103.
Kitchin, J. E. S., Pomeranz, M. K., Pak, G., Washenik, K., & Shupack, J. L. (1997). Rediscovering mycophenolic acid: a review of its mechanism, side effects, and potential uses. Journal of the American Academy of Dermatology, 37(3 Pt 1), 445–449.
Arns, W., Cibrik, D. M., Walker, R. G., Mourad, G., Budde, K., Mueller, E. A., & Vincenti, F. (2006). Therapeutic drug monitoring of mycophenolic acid in solid organ transplant patients treated with mycophenolate mofetil: review of the literature. Transplantation, 82(8), 1004–1012.
Shaw, L. M., Figurski, M., Milone, M. C., Trofe, J., & Bloom, R. D. (2007). Therapeutic drug monitoring of mycophenolic acid. Clinical Journal of the American Society of Nephrology, 2(5), 1062–1072.
Rahman, A. N. A., Tett, S. E., & Staatz, C. E. (2013). Clinical pharmacokinetics and pharmacodynamics of mycophenolate in patients with autoimmune disease. Clinical Pharmacokinetics, 52, 303–331.
Clutterbuck, P. W., Oxford, A. E., Raistrick, H., & Smith, G. (1932). Studies in the biochemistry of micro-organisms: the metabolic products of the Penicillium brevi-compactum series. Biochemical Journal, 26, 1441.
Xu, Z.-N., & Yang, S.-T. (2007). Production of mycophenolic acid by Penicillium brevicompactum immobilized in a rotating fibrous-bed bioreactor. Enzyme and Microbial Technology, 40, 623–628.
Patel, G., Patil, M. D., Soni, S., Khobragade, T. P., Chisti, Y., & Banerjee, U. C. (2016). Production of mycophenolic acid by Penicillium brevicompactum—a comparison of two methods of optimization. Biotechnology Reports, 11, 77–85.
Nguyen, D. T. T., Guillarme, D., Rudaz, S., & Veuthey, J. L. (2006). Fast analysis in liquid chromatography using small particle size and high pressure. Journal of Separation Science, 29(12), 1836–1848.
Alani, F., Grove, J. A., Anderson, W. A., & Moo-Young, M. (2009). Mycophenolic acid production in solid-state fermentation using a packed-bed bioreactor. Biochemical Engineering Journal, 44, 106–110.
Ismaiel, A., Ahmed, A., & El-Sayed, E. (2015). Immobilization technique for enhanced production of the immunosuppressant mycophenolic acid by ultraviolet and gamma-irradiated Penicillium roqueforti. Journal of Applied Microbiology, 119(1), 112–126.
Patel, G., Patil, M. D., Soni, S., Chisti, Y., & Banerjee, U. C. (2017). Production of mycophenolic acid by Penicillium brevicompactum using solid state fermentation. Applied Biochemistry and Biotechnology, 182(1), 97–109.
Vásquez, M. P., Bezerra, M., & Pereira, N. (2006). RSM analysis of the effects of the oxygen transfer coefficient and inoculum size on the xylitol production by Candida guilliermondii. Applied Biochemistry and Biotechnology, 129, 256–264.
Miller, G. L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426–428.
Saran, S., Isar, J., & Saxena, R. K. (2007). Statistical optimization of conditions for protease production fromBacillus sp. and its scale-up in a bioreactor. Applied Biochemistry and Biotechnology, 141(2-3), 229–239.
Himabindu, M., Ravichandra, P., Vishalakshi, K., & Jetty, A. (2006). Optimization of critical medium components for the maximal production of gentamicin by Micromonospora echinospora ATCC 15838 using response surface methodology. Applied Biochemistry and Biotechnology, 134(2), 143–154.
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The authors are thankful to Central Instrument Facility Center (CIFC) and Department of Pharmaceutical Engineering IIT (BHU) Varanasi, India, for providing technical help.
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Anand, S., Srivastava, P. Optimization Strategies for Purification of Mycophenolic Acid Produced by Penicillium brevicompactum. Appl Biochem Biotechnol 191, 867–880 (2020). https://doi.org/10.1007/s12010-019-03204-w
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DOI: https://doi.org/10.1007/s12010-019-03204-w