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Partitioning of Recombinant Pseudomonas putida POS-F84 Proline Dehydrogenase in Aqueous Two-Phase Systems: Optimization Using Response Surface Methodology

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Abstract

Empirical modeling the partition behavior and recovery of a recombinant Pseudomonas putida POS-F84 proline dehydrogenase (ProDH) in aqueous two-phase systems (ATPS) was carried out by response surface methodology (RSM). Polyethylene glycol 1000 (PEG-1000) concentration, sodium carbonate concentration, and pH, which were the most important factors, were chosen for modeling the partition feature of enzyme. The adequacy of the models was investigated by means of variance analysis. Also, to confirm the efficiency of the ATPS in partition and purification of recombinant ProDH, purity and enzymatic activity was studied. After numerical optimization, an optimal ATPS composed of 14.33% PEG-1000 and 11.79% sodium carbonate at pH 7.48 was achieved. Yield, purification factor, and recovery were 81.41%, 60.82, and 270.82%, respectively. Purified recombinant ProDH was found as a single protein band into the upper PEG-rich phase and the specific activity was calculated to be 46.23 ± 2.1 U/mg. Collectively, our data showed that the RSM could be an appropriate and powerful tool to define the best ATPS system for recovery and purification of P. putida ProDH.

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References

  1. Baban, B. A., Vinod, M. P., Tanner, J. J., & Becker, D. F. (2007). Probing a hydrogen bond pair and the FAD redox properties in the proline dehydrogenase domain of Escherichia coli PutA. Biochemical Biophysics Acta, 1701, 49–59.

    Google Scholar 

  2. White, T. A., Krishnan, N., Becker, D. F., & Tanner, J. J. (2007). Structure and kinetics of monofunctional proline dehydrogenase from Thermus thermophilus. Journal of Biological Chemistry, 282(19), 14316–14327.

    Article  CAS  PubMed  Google Scholar 

  3. Shahbaz Mohammadi, H., & Omidinia, E. (2012). Proline dehydrogenase from Pseudomonas fluorescence: gene cloning, purification, characterization and homology modeling. Applied Biochemistry and Microbiology, 48, 191–198.

    Article  CAS  Google Scholar 

  4. Shahbaz Mohammadi, H., & Omidinia, E. (2013). Process integration for the recovery and purification of recombinant Pseudomonas fluorescens proline dehydrogenase using aqueous two-phase systems. Journal of Chromatography B, 929, 11–17.

    Article  CAS  Google Scholar 

  5. Ehliers, T. P., & Wilhelm, J. K. (2011). Polymer phase behavior. New York: Nova Science Publishers, Inc..

    Google Scholar 

  6. Hatti-Kaul, R. (1999). Methods in biotechnology: aqueous two-phase systems: methods and protocols. Totowa: Humana Press Inc..

    Google Scholar 

  7. Dembczynski, R., Białas, W., & Jankowski, T. (2013). Partitioning of lysozyme in aqueous two-phase systems containing ethylene oxide-propylene oxide copolymer and potassium phosphates. Food of Bioproduct Processing, 91(3), 292–302.

    Article  CAS  Google Scholar 

  8. Albertsson, P. A. (1986). Partition of cell particles and macromolecules (3rd ed.). New York: Wiley.

    Google Scholar 

  9. Shahbaz mohamadi, H., & Omidinia, E. (2010). Investigation of chromatography and polymer/salt aqueous two-phase processes for downstream processing development of recombinant phenylalanine dehydrogenase. Bioprocess and Biosystems Engineering, 33, 317–329.

    Article  CAS  PubMed  Google Scholar 

  10. Pérez, R. L., Loureiro, D. B., Nerli, B. B., & Tubio, G. (2015). Optimization of pancreatic trypsin extraction in PEG/citrate aqueous two-phase systems. Protein Expression and Purificiation, 106, 66–71.

    Article  CAS  Google Scholar 

  11. Tatineni, R., Doddapaneni, K. K., Potumarthi, R. C., & Mangamoori, L. (2007). Optimization of keratinase production and enzyme activity using response surface methodology with Streptomyces sp7. Applied Biochemistry and Biotechnology, 141(2-3), 187–201.

    Article  CAS  PubMed  Google Scholar 

  12. Chennupati, S., Potumarthi, R., Gopal Rao, M., Manga, P. L., Sridevi, M., & Jetty, A. (2009). Multiple responses optimization and modeling of lipase production by Rhodotorula mucilaginosa MTCC-8737 using response surface methodology. Applied Biochemistry and Biotechnology, 159(2), 317–329.

    Article  CAS  PubMed  Google Scholar 

  13. Shahbaz Mohammadi, H., Omidinia, E., & Mahdizadehdehosta, R. (2013). Expression, purification and characterization of the proline dehydrogenase domain of PutA from Pseudomonas putida POS-F84. Indian Journal of Microbiology, 53, 297–302.

    Article  CAS  Google Scholar 

  14. Kianmehr, A., Mahrooz, A., Ansari, J., Oladnabi, M., & Shahbazmohammadi, H. (2016). The rapid and sensitive quantitative determination of galactose by combined enzymatic and colorimetric method: application in neonatal screening. Applied Biochemistry and Biotechnology, 179(2), 283–293.

    Article  CAS  PubMed  Google Scholar 

  15. Shahbaz mohammadi, H., & Omidinia, E. (2008). Development and application of aqueous two-phase partition for the recovery and separation of recombinant phenylalanine dehydrogenase. Iranian journal of chemistry and chemical engineering, 28, 119–127.

    Google Scholar 

  16. Liu, X., Mu, T., Sun, H., Zhang, M., & Chen, J. (2013). Optimisation of aqueous two-phase extraction of anthocyanins from purple sweet potatoes by response surface methodology. Food Chemistry, 141(3), 3034–3041.

    Article  CAS  PubMed  Google Scholar 

  17. Bradford, M. M. (1976). Rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principles of protein-dye binding. Analytical Biochemistry, 72(1-2), 248–254.

    Article  CAS  PubMed  Google Scholar 

  18. Sambrook, J., Fritsch, E. F., & Maniatis, T. (1994). Molecular cloning: a laboratory manual (2nd ed.). Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  19. Xu, Q., Shen, Y., Wang, H., Zhang, N., Xu, S., & Zhang, L. (2013). Application of response surface methodology to optimize extraction of flavonoids from fructose sophorae. Food Chemistry, 138(4), 2122–2129.

    Article  CAS  PubMed  Google Scholar 

  20. Tubio, G., Nerli, B., & Pico, G. (2007). Partitioning features of bovine trypsin and alpha chymotrypsin in polyethylene glycol-sodium citrate aqueous two-phase systems. Journal of Chromatography B, 852(1-2), 244–249.

    Article  CAS  Google Scholar 

  21. Yan-Min, L., Yan-Zaho, Y., Xi-Dan, Z., & Chuan-Bo, X. (2010). Bovine serum albumin partitioning in polyethylene glycol (PEG)/potassium citrate aqueous two-phase systems. Food Bioproduct Processing, 88, 40–46.

    Article  CAS  Google Scholar 

  22. Joglekar, A. M., & May, A. T. (1987). Product excellence through design of experiments. Journal of Cereal Foods, 32, 857–868.

    Google Scholar 

  23. Mirhosseini, H., Tan, C. P., Hamid, N. S. A., & Yusof, C. P. (2008). Effect of arabic gum, xanthan gum and orange oil on flavor release from diluted orange beverage emulsion. Food Chemistry, 107, 1161–1172.

    CAS  Google Scholar 

  24. Su, C. K., & Chiang, B. H. (2006). Partitioning and purification of lysozyme from chicken egg white using aqueous two-phase system. Process Biochemistry, 41(2), 257–263.

    Article  CAS  Google Scholar 

  25. Grahovac, J., Dodic, J., Jokic, A., Dodic, S., & Popov, S. (2012). Optimization of ethanol production from thick juice: a response surface methodology approach. Fuel, 93, 221–228.

    Article  CAS  Google Scholar 

  26. Khayet, M., Cojocaru, C., & Zakrzewska-Trznadel, G. (2008). Response surface modeling and optimization in pervaporation. Journal of Membrane Science, 321(2), 272–283.

    Article  CAS  Google Scholar 

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Funding

This project was supported by Young Researchers and Elite Club of Bandar Abbas Branch, Islamic Azad University.

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Authors and Affiliations

  1. Enzyme Technology Laboratory, Department of Biochemistry, Genetic and Metabolism Research Group, Pasteur Institute of Iran, Tehran, Iran

    Eskandar Omidinia, Hamid Shahbazmohammadi & Zeinab MohseniPour

  2. Young Researchers and Elite Club, Bandar Abbas Branch, Islamic Azad University, Bandar Abbas, Iran

    Rahman Mahdizadeh

Authors
  1. Eskandar Omidinia
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  2. Hamid Shahbazmohammadi
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  3. Zeinab MohseniPour
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  4. Rahman Mahdizadeh
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Correspondence to Rahman Mahdizadeh.

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Omidinia, E., Shahbazmohammadi, H., MohseniPour, Z. et al. Partitioning of Recombinant Pseudomonas putida POS-F84 Proline Dehydrogenase in Aqueous Two-Phase Systems: Optimization Using Response Surface Methodology. Appl Biochem Biotechnol 189, 498–510 (2019). https://doi.org/10.1007/s12010-019-03011-3

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  • DOI: https://doi.org/10.1007/s12010-019-03011-3

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