Journals / Journal of the Turkish Chemical Society Section B: Chemical Engineering / 2026 / Volume: 2026

Statistical Optimization of Fermentation Conditions for Sustainable cis,cis-Muconic Acid Biosynthesis by Pseudomonas putida

Emine Bayraktar(ANKARA ÜNİVERSİTESİ, MÜHENDİSLİK FAKÜLTESİ)0000-0003-1947-3804
Pages
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Publication Date
2026-03-21
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

The results showed that ccMA production was most influenced by the initial pH of the medium and concentrations of Na-benzoate, NaCl, and (NH4)2SO4.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Many chemicals used in industry are produced using different isomers of muconic acid.
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Key Findings & Data:Statistical experimental design was applied to optimize the production of ccMA by Pseudomonas putida KT2440 in batch culture.
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Conclusion & Impact:The results showed that ccMA production was most influenced by the initial pH of the medium and concentrations of Na-benzoate, NaCl, and (NH4)2SO4.
🔬Design:Modeling & Simulation
📍Scope:Comprehensive Sample
⚙️Variables:cis-cis muconic acid · Pseudomonas putida · fermentation

Abstract

Many chemicals used in industry are produced using different isomers of muconic acid. The isomers are especially precursors for many industrially important polymers. In this study, eco-friendly production of cis, cis-muconic acid (ccMA) used in the production of polymers was carried out. Statistical experimental design was applied to optimize the production of ccMA by Pseudomonas putida KT2440 in batch culture. Plackett-Burman design was used to investigate the effect of 11 variables for ccMA production in the first optimization step. The results showed that ccMA production was most influenced by the initial pH of the medium and concentrations of Na-benzoate, NaCl, and (NH4)2SO4. After these significant factors were determined, the optimization of these factors was carried out with Response Surface Methodology (RSM) as the second optimization step. RSM showed that the optimal values of the factors (the initial pH and concentrations of Na-benzoate, NaCl, and (NH4)2SO4) were 7.96, 16.83 mM, 0.7 and 1.4 g/L, respectively. The other components of the fermentation medium were 18 g/L Na2HPO4, 10 g/L KH2PO4, 20 mM glucose, 1 mM MgSO4, 150 µM CaCl2, 13 µM FeSO4, and temperature of 35 oC. ccMA production increased about 4 times using statistical optimization methods.

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Cite This Article

Uzun, F. E., Songür, R., Bayraktar, E. (2026). Statistical Optimization of Fermentation Conditions for Sustainable cis,cis-Muconic Acid Biosynthesis by Pseudomonas putida. Journal of the Turkish Chemical Society Section B: Chemical Engineering, Volume: 2026. https://doi.org/10.58692/jotcsb.1880143

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References (27)

  1. Arumugam, T., & Senthil Kumar, P. (2017). Optimization of media components for production of antimicrobial compound by Brevibacillus brevis EGS9 isolated from mangrove ecosystem. Journal of Microbiological Methods, 142, 83–89. https://doi.org/10.1016/j.mimet.2017.09.010
  2. Bayraktar, E. (2001). Response surface optimization of the separation of DL-tryptophan using an emulsion liquid membrane. Process Biochemistry, 37, 169–175. www.elsevier.com/locate/procbio
  3. Bezerra, M. A., Santelli, R. E., Oliveira, E. P., Villar, L. S., & Escaleira, L. A. (2008). Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta, 76, 965–977. https://doi.org/10.1016/j.talanta.2008.05.019
  4. Choi, S., Lee, H. N., Park, E., Lee, S. J., & Kim, E. S. (2020). Recent advances in microbial production of cis,cis-muconic acid. Biomolecules, 10(9), 1–14. https://doi.org/10.3390/biom10091238
  5. Dholariya, Y., Bansod, Y., Vora, R., Mittal, S., Shirsat, A., & Bhingare, C. (2014). Design and optimization of bilayered tablet of Hydrochlorothiazide using the Quality-by-Design approach. International Journal of Pharmaceutical Investigation, 4(2), 93–101. https://doi.org/10.4103/2230-973x.133058
  6. Eisa, M., El-Refai, H., & Amin, M. (2016). Single step biotransformation of corn oil phytosterols to boldenone by a newly isolated Pseudomonas aeruginosa. Biotechnology Reports, 11, 36–43. https://doi.org/10.1016/j.btre.2016.05.002
  7. Gonabadi, E., Samadlouie, H. R., & Shafafi Zenoozian, M. (2021). Optimization of culture conditions for enhanced Dunaliella salina productions in mixotrophic culture. Preparative Biochemistry and Biotechnology, 1–9. https://doi.org/10.1080/10826068.2021.1922917
  8. Gorden, J., Zeiner, T., & Brandenbusch, C. (2015). Reactive extraction of cis, cis -muconic acid. Fluid Phase Equilibria, 393, 78–84.
  9. Gu Bang, S., Jae Choi, W., Yong Choi, C., & Hwan Cho, M. (1996). Production of cis, cis-Muconic Acid from Benzoic Acid via Microbial Transformation. Biotechnol. Bioprocess Eng, 1, 36–40.
  10. Johnson, C. W., Salvachúa, D., Khanna, P., Smith, H., Peterson, D. J., & Beckham, G. T. (2016). Enhancing muconic acid production from glucose and lignin-derived aromatic compounds via increased protocatechuate decarboxylase activity. Metabolic Engineering Communications, 3, 111–119. https://doi.org/10.1016/j.meteno.2016.04.002
  11. Kumar, R., Mahajan, S., Kumar, A., & Singh, D. (2011). Identification of variables and value optimization for optimum lipase production by Bacillus pumilus RK31 using statistical methodology. New Biotechnology, 28(1), 65–71. https://doi.org/10.1016/j.nbt.2010.06.007
  12. Li, M., Eskridge, K., Liu, E., & Wilkins, M. (2019). Enhancement of polyhydroxybutyrate (PHB) production by 10-fold from alkaline pretreatment liquor with an oxidative enzyme-mediator-surfactant system under Plackett-Burman and central composite designs. Bioresource Technology, 281, 99–106. https://doi.org/10.1016/j.biortech.2019.02.045
  13. Mu, X. Q., Xu, Y., Nie, Y., Ouyang, J., & Sun, Z. H. (2005). Candida parapsilosis CCTCC M203011 and the optimization of fermentation medium for stereoinversion of (S)-1-phenyl-1,2-ethanediol. Process Biochemistry, 40(7), 2345–2350. https://doi.org/10.1016/j.procbio.2004.09.026
  14. Murthy, M. S. R. C., Swaminathan, T., Rakshit, S. K., & Kosugi, Y. (2000). Statistical optimization of lipase catalyzed hydrolysis of methyloleate by response surface methodology. Bioprocess Engineering, 22, 35–39.
  15. Pawar, S., & Chaudhari, A. (2020). Pyrrolnitrin from Rhizospheric Serratia marcescens NCIM 5696: Optimization of Process Parameters Using Statistical Tools and Seed-Applied Bioprotectants for Vigna radiata (L.) Against Fusarium oxysporum MTCC 9913. Applied Biochemistry and Biotechnology, 190(3), 803–825. https://doi.org/10.1007/s12010-019-03123-w
  16. Plackett, R. L., & Burman, J. P. (1946). The Design of Optimum Multifactorial Experiments. Biometrika, 33(4), 305–325.
  17. Rajendran, A., & Thangavelu, V. (2009). Statistical experimental design for evaluation of medium components for lipase production by Rhizopus arrhizus MTCC 2233. LWT- Food Science and Technology, 42(5), 985–992. https://doi.org/10.1016/j.lwt.2008.12.009
  18. Scelfo, S., Pirone, R., & Russo, N. (2016). Highly efficient catalysts for the synthesis of adipic acid from cis,cis-muconic acid. Catalysis Communications, 84, 98–102. https://doi.org/10.1016/j.catcom.2016.06.013
  19. Sudarsan, S., Blank, L. M., Dietrich, A., Vielhauer, O., Takors, R., Schmid, A., & Reuss, M. (2016). Dynamics of benzoate metabolism in Pseudomonas putida KT2440. Metabolic Engineering Communications, 3, 97–110. https://doi.org/10.1016/j.meteno.2016.03.005
  20. Van Duuren, J. B. J. H., Wijte, D., Karge, B., Martins dos Santos, V. A. P., Yang, Y., Mars, A. E., & Eggink, G. (2012). PH-stat fed-batch process to enhance the production of cis, cis-muconate from benzoate by Pseudomonas putida KT2440-JD1. Biotechnology Progress, 28(1), 85–92. https://doi.org/10.1002/btpr.709
  21. Vardon, D. R., Rorrer, N. A., Salvachúa, D., Settle, A. E., Johnson, C. W., Menart, M. J., Cleveland, N. S., Ciesielski, P. N., Steirer, K. X., Dorgan, J. R., & Beckham, G. T. (2016). Cis, cis -Muconic acid: Separation and catalysis to bio-adipic acid for nylon-6,6 polymerization. Green Chemistry, 18(11), 3397–3413. https://doi.org/10.1039/c5gc02844b
  22. Wu, C. M., Wu, C. C., Su, C. C., Lee, S. N., Lee, Y. A., & Wu, J. Y. (2006). Microbial synthesis of cis,cis-muconic acid from benzoate by Sphingobacterium sp. mutants. Biochemical Engineering Journal, 29, 35–40. https://doi.org/10.1016/j.bej.2005.02.034
  23. Wu, C., Wang, L., Li, H., & Yu, S. (2020). Combination of solid-phase extraction with microextraction techniques followed by HPLC for simultaneous determination of 2-methylimidazole and 4-methylimidazole in beverages. Food Chemistry, 305, 125389. https://doi.org/10.1016/j.foodchem.2019.125389
  24. Xie, N. Z., Liang, H., Huang, R. B., & Xu, P. (2014). Biotechnological production of muconic acid: Current status and future prospects. Biotechnology Advances, 32, 615–622. https://doi.org/10.1016/j.biotechadv.2014.04.001
  25. Xie, N. Z., Wang, Q. Y., Zhu, Q. X., Qin, Y., Tao, F., Huang, R. B., & Xu, P. (2014). Optimization of medium composition for cis,cis-muconic acid production by a Pseudomonas sp. mutant using statistical methods. Preparative Biochemistry and Biotechnology, 44(4), 342–354. https://doi.org/10.1080/10826068.2013.829497
  26. Yang, H., Lin, X., Zhong, X., Cao, M., Yuan, J., Li, Z., Ling, X., & He, N. (2025). Current status and advances in the green synthesis of muconic acid. In Critical Reviews in Biotechnology (Vol. 45, Number 5, pp. 1040–1057). Taylor and Francis Ltd. https://doi.org/10.1080/07388551.2024.2433998
  27. Yoo, H. Y., Yang, X., Kim, D. S., Lee, S. K., Lotrakul, P., Prasongsuk, S., Punnapayak, H., & Kim, S. W. (2016). Evaluation of the overall process on bioethanol production from miscanthus hydrolysates obtained by dilute acid pretreatment. Biotechnology and Bioprocess Engineering, 21(6), 733–742. https://doi.org/10.1007/s12257-016-0485-x