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Thermodynamic study of lysozyme adsorption on cation-exchange monolithic adsorbent

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Abstract

The present study aimed to produce a macroporous monolithic cation-exchanger and evaluate the effect of temperature on the thermodynamic behavior of lysozyme adsorption on it. The adsorbent matrix was made from the catalyzed polymerization of acrylamide and bis-acrylamide and functionalized with acrylic acid grafting. The exchanger was characterized by scanning electron microscopy (SEM), ion exchange capacity (IEC), point of zero charges (PZC), infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and X-ray diffraction (XRD). The adsorption of egg white lysozyme (LYS) onto the adsorbent was evaluated at various temperatures (283.15 to 323.15 K), and a maximum adsorption capacity was observed at 313.15 K, corresponding to ~ 1055 mg LYS.g−1. The adsorption isotherms were determined by the batch method at pH 7.2, and initial lysozyme concentrations ranging from 0.1 to 20.0 mg·mL−1. Langmuir model was adjusted to the adsorption equilibrium data and nonlinear Van’t Hoff's method was used to determine thermodynamic parameters. An increase in adsorption capacity was observed with the increase in temperature up to 313.15 K. Thermodynamic analysis showed that the process was spontaneous, with enthalpy-entropy compensation under the conditions studied, and the ion exchange process was entropy-driven as the temperature has increased. Finally, the produced matrix was tested for the partial purification of lysozyme from chicken egg white, reaching a production yield of 38% and a purification factor of 2.1 in a single chromatographic step. Thus, indicating that the matrix has potential in purification processes by cation exchange.

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References

  • Akkaya B, Akkaya R (2012) A crosslinked carboxylic acid containing cation exchange monolithic cryogel for human serum albumin separation. J Macromol Sci 49:736–743

    Article  CAS  Google Scholar 

  • Bigman LS, Levy Y (2018) Entropy-enthalpy compensation in conjugated proteins. Chem Phys 514:95–105

    Article  CAS  Google Scholar 

  • Bonomo RCF, Minim LA, Fontan RCI, Coimbra JSR (2006) Hydrophobic interaction adsorption of whey proteins: effect of temperature and salt concentration and thermodynamic analysis. J Chromatogr B 844(1):6–14

    Article  CAS  Google Scholar 

  • Bradford MM (1976) Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Anal Biochem 72(1–2):248–254

    Article  CAS  PubMed  Google Scholar 

  • Bregado JL, Secchi AR, Tavares FW, Rodrigues DS, Gambetta R (2019) Amorphous paracrystalline structures from native crystalline cellulose: a molecular dynamics protocol. Fluid Phase Equilib 491:56–76

    Article  CAS  Google Scholar 

  • Carvalho BMA, Silva SLD, Silva LHM, Minim VPR, Silva MCH, Carvalho LM, Minim LA (2014) Cryogel poly (acrylamide): synthesis, structure and applications. Sep Purif Rev 43:241–262

    Article  CAS  Google Scholar 

  • Da Silva JF, Da Silva DL, Nascimento RG, Veríssimo LAA, Veloso CM, Bonomo RCF, Fontan RCI (2019) Enhancements in sugar immobilization in polymeric macroporous matrices for affinity capture. J Appl Polym Sci 136(37):47956

    Article  Google Scholar 

  • De Oliveira ACF, Neves ICO, Saraiva JAM, De Carvalho MFF, Batista GA, Veríssimo LAA, De Vilela JV (2019) Capture of lysozyme on macroporous cryogels by hydrophobic affinity chromatography. Separ Sci Techonol 55(11):2012–2024

    Article  Google Scholar 

  • Dragan AI, Read CM, Crane-Robinson C (2017) Enthalpy–entropy compensation: the role of solvation. Eur Biophys J 46(4):301–308

    Article  CAS  PubMed  Google Scholar 

  • Ertürk G, Mattiasson B (2014) Cryogels-versatile tools in bioseparation. J Chromatogr A 1357:24–35

    Article  PubMed  Google Scholar 

  • Fontan RCI, Bonomo RCF, Gonçalves GRF, Minim VPR, Minim LA (2018) Alternatives for characterizing macroporous polyacrylamide monolithic ion exchanger columns. Polym Eng Sci 58(10):1717–1725

    Article  CAS  Google Scholar 

  • García E, Rodriguez L, Ferro V, Valverde JL (2019) Prediction of multicomponent ION exchange equilibria by using the e-NRTL model for computing the activity coefficients in solution. Fluid Phase Equilibria 498:132–143

    Article  Google Scholar 

  • Ghalandari V, Hashemipoura H, Bagheria H (2020) Experimental and modeling investigation of adsorption equilibrium of CH4, CO2, and N2 on activated carbon and prediction of multi-component adsorption equilibrium. Fluid Phase Equilib 508:12–119

    Article  Google Scholar 

  • Gonçalves GRF, Gandolfi ORR, Santos CMS, Bonomo RCF, Veloso CM, Fontan RCI (2016) Development of supermacroporous monolithic adsorbents for purifying lectins by affinity with sugars. J Chromatogr B 1033:406–412

    Article  Google Scholar 

  • Gonçalves GRF, Gandolfi ORR, Santos LS, Bonomo RCF, Veloso CM, Veríssimo LAA, Fontan RDCI (2017) Immobilization of sugars in supermacroporous cryogels for the purification of lectins by affinity chromatography. J Chromatogr B 1068:71–77

    Article  Google Scholar 

  • Guan YF, Lai SY, Lin CS, Suen SY, Wang MY (2019) Purification of lysozyme from chicken egg white using diatom frustules. Food Chem 286:483–490

    Article  CAS  PubMed  Google Scholar 

  • Kotsalos E, Sevastos D, Karaiskakis G (2019) Variation of linear alcohols’ activity coefficients, referred to aqueous solutions of methanol, ethanol and 1-propanol, in the presence of various types of surfactants, studied by reversed – Flow Gas Chromatography. Fluid Phase Equilib 495:69–75

    Article  CAS  Google Scholar 

  • Kumar A, Bansal V, Andersson J, Roychoudhury PK, Mattiasson B (2006) Supermacroporous cryogel matrix for integrated protein isolation immobilized metal affinity chromatographic purification of urokinase from cell broth of a human kidney cell line. J Chromatogr A 1103:35–42

    Article  CAS  PubMed  Google Scholar 

  • Langmuir I (1916) The dissociation of hydrogen into atoms. III. The mechanism of the reaction. J Am Chem Soc 38(6):1145–1156

    Article  CAS  Google Scholar 

  • Mól PCG, Veríssimo LAA, Eller MR, Minim VPR, Minim LA (2017) Development of an affinity cryogel for one step purification of lysozyme from chicken egg white. J Chromatogr B 1044:17–23

    Article  Google Scholar 

  • Mourão CA, Marcuz C, Haupt K, Bueno SMA (2019) Polyacrylamide-alginate (PAAm-Alg) and phospho-L-tyrosine-linked PAAm-Alg monolithic cryogels: purification of IgG from human serum. J Chromatogr B 1129:559–567

    Article  Google Scholar 

  • Nascimento IS, Silva LD, Verissimo LAA, Pereira TB, Goncalves GRF, Veloso CM, Bonomo RFC, Fontan RCI (2019) Single-step purification of lectins from jackfruit (Artocarpus integrifolia) seeds using a supermacroporous ion exchange cryogel. Rev Mex Ing Quim 18:313–324

    Google Scholar 

  • Paiva TS, Nascimento RG, Veríssimo LAA, Bonomo RCF, Veloso CM, Resende JV, Fontan, RDCI (2021) Adsorption isotherms and thermodynamic properties of a butyl functionalized hydrophobic macroporous cryogel. Brazilian J Chem Eng 1–9

  • Plieva FM, Andersson J, Galaev IV, Mattiasson B, Savina IN, Deraz S (2004a) Characterization of supermacroporous monolithic polyacrylamide based matrices designed for chromatography of bioparticles. J Chromatogr B 807(1):129–137

    Article  CAS  Google Scholar 

  • Plieva FM, Andersson J, Galaev IY, Mattiasson B (2004b) Characterization of polyacrylamide based monolithic columns. J Sep Sci 27(10–11):828–836

    Article  CAS  PubMed  Google Scholar 

  • Privar Y, Malakhova I, Pestov A, Fedorets A, Azarova Y, Schwarz S, Bratskaya S (2018) Polyethyleneimine cryogels for metal ions sorption. Chem Eng J 334:1392–1398

    Article  CAS  Google Scholar 

  • Ren CS, Wang DZ, Wang YN (2006) Graft co-polymerization of acrylic acid onto the linen surface induced by DBD in air. Surf Coat Technol 201:2867–2870

    Article  CAS  Google Scholar 

  • Rodler A, Ueberbacher R, Beyer B, Jungbauer A (2019) Calorimetry for studying the adsorption of proteins in hydrophobic interaction chromatography. Prep Biochem Biotechnol 49(1):1–20

    Article  CAS  PubMed  Google Scholar 

  • Sahiner N, Demirci S (2016) Conducting semi-interpenetrating polymeric composites via the preparation of poly (aniline), poly (thiophene), and poly (pyrrole) polymers within superporous poly (acrylic acid) cryogels. React Funct Polym 105:60–65

    Article  CAS  Google Scholar 

  • Sahiner N, Yildiz S, Sahiner M, Aa I, Al-Lohedan H (2015) Macroporous cryogel metal nanoparticle composites for H2 generation from NaBH4 hydrolysis in seawater. Appl Surf Sci 354:388–396

    Article  CAS  Google Scholar 

  • Savina IN, Mattiasson B, Galaev IY (2005) Graft polymerization of acrylic acid onto macroporous polyacrylamide gel (cryogel) initiated by potassium diperiodatocuprate. Polymer 46(23):9596–9603

    Article  CAS  Google Scholar 

  • Shugar D (1952) The measurement of lysozyme activity and the ultra-violet inactivation of lysozyme. Biochem Biophys Acta 8:302–309

    Article  CAS  PubMed  Google Scholar 

  • Teske CA, Blanch HW, Prausnitza JM (2004) Chromatographic measurement of interactions between unlike proteins. Fluid Fhase Equilibira 219:139–148

    Article  CAS  Google Scholar 

  • Ueberbacher R, Rodler A, Hahn R, Jungbauer A (2010) Hydrophobic interaction chromatography of proteins: Thermodynamic analysis of conformational changes. J Chromatogr A 1217(2):184–190

    Article  CAS  PubMed  Google Scholar 

  • van Stokkum IHM, Linsdell H, Hadden JM, Haris PI, Chapman D, Bloemendal M (1995) Temperature-induced changes in protein structures studied by fourier transform infrared spectroscopy and global analysis. Biochemistry 34:10508

    Article  PubMed  Google Scholar 

  • Veríssimo LAA, Paganoto FS, Mol PCG, Fontan RDCI, Minim VPR, Minim LA (2017) Preparation of an affinity cryogel column for lysozyme purification. Sep Sci Technol 52:1973–1982

    Article  Google Scholar 

  • Vieira ED, Basso LGM, Costa-Filho AJ (2017) Non-linear vant Hoff behavior in pulmonary surfactant model membranes. Biochimica et Biophysica Acta (BBA) Biomembranes 1859(6):1133–1143

  • Watanabe EO, Popova E, Miranda EA, Maurer G (2009) Phase equilibria for salt-induced lysozyme precipitation: Effect of salt type and temperature. Fluid Phase Equilibira 281:32–39

    Article  CAS  Google Scholar 

  • Xia YQ, Guo TY, Song MD, Zhang BH, Zhang BL (2005) Hemoglobin recognition by imprinting in semi-interpenetrating polymer network hydrogel based on polyacrylamide and chitosan. Biomacromol 6:2601–2606

    Article  CAS  Google Scholar 

  • Yan L, Shen S, Yun J, Yao K (2011) Isolation of Lysozyme from Chicken Egg White Using Polyacrylamide-based Cation-exchange Cryogel. Chin J Chem Eng 19(5):876–880. https://doi.org/10.1016/S1004-9541(11)60068-2

  • Yao K, Shen S, Yun J, Wang L, He X, Yu X (2006) Characterization of a novel continuous supermacroporous monolithic cryogel embedded with nanoparticles for protein chromatography. J Chromatogr A 1109(1):103–110

    Article  CAS  PubMed  Google Scholar 

  • Yao K, Yun J, Shen S, Chen F (2007) In-situ graft-polymerization preparation of cation-exchange supermacroporous cryogel with sulfo groups in glass columns. J Chromatogr A 1157:246–251

    Article  CAS  PubMed  Google Scholar 

  • Zhao Y, Li X, Yu L, Dong X, Sun Y (2019) Lysozyme adsorption to cation exchanger derivatized by sequential modification of poly (ethylenimine)-Sepharose with succinic anhydride and ethanolamine: Effect of pH and ionic strength. Chin J Chem Eng 34:87–94

    Google Scholar 

Download references

Funding

This work was supported by the State University of Southwest of Bahia (UESB) by labs access and Aux-PPG Program, Coordination for the Improvement of Higher Education Personnel (CAPES) by funding 819178/2015 and 88881.068456/2014-01 and National Council for Scientific and Technological Development (CNPq) by funding 314071/2021-0.

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Authors and Affiliations

  1. Process Engineering Laboratory, State University of Southwest of Bahia, Itapetinga, BA, 45700-000, Brazil

    Yara Gomes de Souza, Izabella de Carvalho Batista Muniz, Renata Cristina Ferreira Bonomo & Rafael da Costa Ilhéu Fontan

  2. Federal Institute of Education, Science and Technology of Pará, Campus Marabá, Marabá, PA, Brazil

    Rui Gomes Nascimento

Authors
  1. Yara Gomes de Souza
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  2. Rui Gomes Nascimento
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  3. Izabella de Carvalho Batista Muniz
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  4. Renata Cristina Ferreira Bonomo
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  5. Rafael da Costa Ilhéu Fontan
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by YGS, RGN and ICBM. The first draft of the manuscript was written by YGS, RCFB and RCIF and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Rafael da Costa Ilhéu Fontan.

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de Souza, Y.G., Nascimento, R.G., de Carvalho Batista Muniz, I. et al. Thermodynamic study of lysozyme adsorption on cation-exchange monolithic adsorbent. Braz. J. Chem. Eng. 41, 519–532 (2024). https://doi.org/10.1007/s43153-023-00373-4

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  • DOI: https://doi.org/10.1007/s43153-023-00373-4

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