Purification of ε-poly-L-lysine from microbial fermentation broth using ion-exchange chromatography in a liquid-circulating fluidized bed

Keywords

ε-poly-L-lysine / Liquid-circulating fluidized bed / Ion-exchange chromatography / Biomanufacturing

References

[1] Bahadır Acıkara Ö. Ion-exchange chromatography and its applications [M]//MARTIN D F, MARTIN D. Column chromatography, 2013, IntechOpen. Rijeka

[2] Baur D, Angelo JM, Chollangi S, et al. . Model assisted comparison of protein a resins and multi-column chromatography for capture processes. J Biotechnol, 2018, 285: 64-73

[3] Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 1976;72:248–54.

[4] Chen J, Li M, He W, et al. . Facile organocatalyzed synthesis of poly(ε-lysine) under mild conditions. Macromolecules, 2017, 50: 9128-34

[5] Chen K-H, Wang SSS, Show P-L, et al. . Rapid and efficient recovery of c-phycocyanin from highly turbid spirulina platensis algae using stirred fluidized bed ion exchange chromatography. Sep Purif Technol, 2019, 209: 636-45

[6] Chen XS, Diao WJ, Ma Y, et al. . Extraction and purification of ε-poly-l-lysine from fermentation broth using an ethanol/ammonium sulfate aqueous two-phase system combined with ultrafiltration. RSC Adv, 2020, 10: 29587-93

[7] Chen X-S, Gao Y, Zhen B et al. Separation and purification of ϵ-poly-l-lysine from fermentation broth. Process Biochem, 2016;51134 – 41. https://doi.org/10.1016/j.procbio.2015.11.010

[8] Chen XS, Li Q, He HG, et al. . Effect of ion form of the ion-exchange resin on ε-poly-l-lysine purification from microbial fermentation broth. RSC Adv, 2019, 9: 12174-81

[9] Ding L, Deng H, Wu C, et al. Affecting factors, equilibrium, kinetics and thermodynamics of bromide removal from aqueous solutions by miex resin. Chem Eng J. 2012;181–2. https://doi.org/10.1016/j.cej.2011.11.096

[10] Ding WY, Guo YG, Zhang HJ, et al. . Efficient purification of the antimicrobial peptide c-poly-l-lysine from microbial culture medium with continuous ion-exchange chromatography. Sep Purif Technol, 2024, 339: 13

[11] Foo KY, Hameed BH. Insights into the modeling of adsorption isotherm systems. Chem Eng J, 2010, 156: 2-10

[12] Gupta SS, Mishra V, Mukherjee MD et al. Amino acid derived biopolymers: recent advances and biomedical applications. Int J Biol Macromol, 2021;188:542 – 67. https://doi.org/10.1016/j.ijbiomac.2021.08.036

[13] Han X, Wang Q, Mi J, et al. . Ε-polylysine-mediated assembly regulating the band gap of natural polyphenols for boosted photodynamic antimicrobial effect. Chem Eng J, 2025, 507: 160420

[14] Ho YS, McKay G. Pseudo-second order model for sorption processes. Process Biochem 1999;34:451 – 65.

[15] Itzhaki RF. Colorimetric method for estimating polylysine and polyarginine. Anal Biochem. 1972;50:569–74. .

[16] Kalaga DV, Dhar A, Dalvi SV et al. Particle-liquid mass transfer in solid–liquid fluidized beds. Chem Eng J, 2014;245:323 – 41.https://doi.org/10.1016/j.cej.2014.02.038

[17] Kang S, Lee B, Ahn K-H, et al. . Facile synthesis of copper-substituted prussian blue analog immobilized ion exchange resins for high-performance ammonium recovery from wastewater: Adsorption kinetics, isotherms, and regeneration. Chem Eng J, 2023, 457: 141128

[18] Karau A, Benken C, Thömmes J, et al. . The influence of particle size distribution and operating conditions on the adsorption performance in fluidized beds. Biotechnol Bioeng, 1997, 55: 54-64

[19] Khadir A, Motamedi M, Negarestani M, et al. . Preparation of a nano bio-composite based on cellulosic biomass and conducting polymeric nanoparticles for ibuprofen removal: Kinetics, isotherms, and energy site distribution. Int J Biol Macromol, 2020, 162: 663-77

[20] Li K, Chen M, Chen L, et al. . Adsorption of tetracycline from aqueous solution by zif-8: Isotherms, kinetics and thermodynamics. Environ Res, 2024, 241: 117588

[21] Lima EC, Hosseini-Bandegharaei A, Moreno-Piraján JC, et al. . A critical review of the estimation of the thermodynamic parameters on adsorption equilibria. Wrong use of equilibrium constant in the van’t hoof equation for calculation of thermodynamic parameters of adsorption. J Mol Liq, 2019, 273: 425-34

[22] Li Q, Chen Q, Yang K, et al. . Ε-poly-l-lysine/rhamnolipid bifunctional coating based on green electrostatic assembly technology for fruit packaging and preservation. Chem Eng J, 2025, 503: 158425

[23] Li S, Mao Y, Zhang L, et al. . Recent advances in microbial ε-poly-l-lysine fermentation and its diverse applications. Biotechnol Biofuels Bioprod, 2022, 15: 65

[24] Li S, Shi C, Ai H, et al. . Bacterial cellulose-based submicrobundle framework-induced hyperporous cellulose microspheres functionalized by ionic liquids for anion exchange chromatography. Chem Eng J, 2023, 475: 146030

[25] Liu J-N, Chang S-L, Xu P-W, et al. . Structural changes and antibacterial activity of epsilon-poly-l-lysine in response to ph and phase transition and their mechanisms. J Agric Food Chem, 2020, 68: 1101-9

[26] Liu Y. Is the free energy change of adsorption correctly calculated?. J Chem Eng Data, 2009, 54: 1981-5

[27] Lv B, Zhao Z, Deng X, et al. . Hydrodynamics and adsorption performance of liquid–solid fluidized bed with granular activated carbon for removal of copper ions from wastewater. J Clean Prod, 2021, 328: 129627

[28] Maciel KS, Santos LS, Bonomo RCF, et al. . Purification of lactoferrin from sweet whey using ultrafiltration followed by expanded bed chromatography. Sep Purif Technol, 2020, 251: 117324

[29] Maliyekkal SM, Shukla S, Philip L et al. Enhanced fluoride removal from drinking water by magnesia-amended activated alumina granules. Chem Eng J, 2008;140:183 – 92. https://doi.org/10.1016/j.cej.2007.09.049

[30] Milonjic SK. A consideration of the correct calculation of thermodynamic parameters of adsorption. J Serb Chem Soc, 2007, 72: 1363-7

[31] Mohamed Nasser S, Abbas M, Trari M. Understanding the rate-limiting step adsorption kinetics onto biomaterials for mechanism adsorption control. Prog React Kinet Mech, 2024, 49: 14686783241226858

[32] Mozaffari Majd M, Kordzadeh-Kermani V, Ghalandari V, et al. . Adsorption isotherm models: a comprehensive and systematic review (2010 – 2020). Sci Total Environ, 2022, 812: 151334

[33] Naidu H, Mathews AP. Linear driving force analysis of adsorption dynamics in stratified fixed-bed adsorbers. Sep Purif Technol, 2021, 257: 117955

[34] Neji M, Bary B, Le Bescop P et al. Swelling behavior of ion exchange resins incorporated in tri-calcium silicate cement matrix: I. Chemical analysis. Journal of Nuclear Materials, 2015;467:544 – 56.

[35] Nijssen TMJ, Padding JT, Ottens M. Hydrodynamics of expanded bed adsorption studied through cfd-dem. Chem Eng Sci, 2023, 280: 119027

[36] Ren P, Li W, Yu K. Computational fluid dynamics simulation of adsorption process in a liquid-solids fluidized bed. J Environ Chem Eng, 2021, 9: 105428

[37] Sethi P, Barman S, Basu S. Strategic tuning of go ratios in cubtc-go nanocomposites for next-generation tetracycline adsorption: a deep dive into isotherms, kinetics, and thermodynamics. Sep Purif Technol, 2025, 361: 131311

[38] Song CP, Ooi CW, Tey BT, et al. . Direct recovery of enhanced green fluorescent protein from unclarified < i>escherichia coli homogenate using ion exchange chromatography in stirred fluidized bed. Int J Biol Macromol, 2020, 164: 4455-65

[39] Sui Y, Yang T, Zhao Y, et al. . Purification of ε-polylysine from fermentation broth by a new method of metal ion complexation. Process Biochem, 2023, 130: 512-9

[40] Terada K, Tsuchiya K, Lamprou A, et al. . Chemoenzymatic synthesis of poly-l-lysine via esterification with alcohol in one-pot. ACS Polym Au, 2025, 5: 26-34

[41] Thömmes J. Fluidized bed adsorption as a primary recovery step in protein purification. New Enzymes for Organic Synthesis. Screening, Suppl Eng 2007;185–230.

[42] Tran HN, You SJ, Hosseini-Bandegharaei A, et al. . Mistakes and inconsistencies regarding adsorption of contaminants from aqueous solutions: a critical review. Water Res, 2017, 120: 88-116

[43] Wang E-Z, Yang Z-M, Guo S-N, et al. . Engineered ε-polylysine hydrochloride aerogel with tailored hierarchical structure and affinity sites for efficient iodide removal from concentrated algae juice. Chem Eng J, 2024, 500: 156818

[44] Wang J, Guo X. Rethinking of the intraparticle diffusion adsorption kinetics model: Interpretation, solving methods and applications. Chemosphere, 2022, 309: 136732

[45] Wang JL, Guo X. Adsorption kinetic models: Physical meanings, applications, and solving methods. J Hazard Mater, 2020, 390: 18

[46] Wang L, Zhang C, Zhang J, et al. . Epsilon-poly-l-lysine: Recent advances in biomanufacturing and applications. Front Bioeng Biotechnol, 2021, 9: 748976

[47] Wang Y, Wang C, Huang X, et al. . Guideline for modeling solid-liquid adsorption: kinetics, isotherm, fixed bed, and thermodynamics. Chemosphere, 2024, 349: 140736

[48] Yamanaka K, Maruyama C, Takagi H, et al. . Epsilon-poly-l-lysine dispersity is controlled by a highly unusual nonribosomal peptide synthetase. Nat Chem Biol, 2008, 4: 766-72

[49] Yousef RI, El-Eswed B, Al-Muhtaseb AH. Adsorption characteristics of natural zeolites as solid adsorbents for phenol removal from aqueous solutions: Kinetics, mechanism, and thermodynamics studies. Chem Eng J, 2011;171:1143-9. https://doi.org/10.1016/j.cej.2011.05.012

[50] You X, Wang Z, Wang L, et al. . Graphene oxide/ε-poly-l-lysine self-assembled functionalized coatings improve the biocompatibility and antibacterial properties of titanium implants. Front Bioeng Biotechnol, 2024, 12: 1381685

[51] Yu H, Liu L, Li X, et al. . Fabrication of polylysine based antibacterial coating for catheters by facile electrostatic interaction. Chem Eng J, 2019, 360: 1030-41

[52] Yu Y, Xia L, Wen J. Fermentation optimization and metabolomic analysis of ε-polylysine production in streptomyces albulus. Biochem Eng J, 2025, 220: 109758

[53] Zhen B, Chen X, Han D, et al. . An alternative method for the decoloration of ɛ-poly-l-lysine eluate by macroporous resin in the separation and purification of ɛ-poly-l-lysine from fermentation broth. Food Bioprod Process, 2015, 95: 332-8

[54] Zhou X, Sun Y, He W, et al. . Solid-liquid phase microextractive adsorption of bio-lactic acid by a novel microextractor immobilizing ionic liquid a336. Chem Eng J, 2024, 500: 157296

[55] Zhu D, Zhang J, Li S et al. Enhancing ε-poly-l-lysine production in streptomyces albulus through l-lysine importer engineering. Metabolic Engineering, 2025;92:102 – 12. https://doi.org/10.1016/j.ymben.2025.07.010

[56] Zhu M, Zhang Z, Liu Y, et al. . Optimization of process parameters for ε -polylysine production by response surface methods. Int J Polym Sci, 2016, 2016: 1-11

[57] Zhu Q, Moggridge GD, D’Agostino C. Adsorption of pyridine from aqueous solutions by polymeric adsorbents mn 200 and mn 500. Part 2: Kinetics and diffusion analysis. Chem Eng J, 2016, 306: 1223-33

[58] Zong H, He Y, Zhan Y, et al. . Optimization of medium constituents for ε-poly-l-lysine fermentation with response surface methodology. J Food Sci, 2010, 75: M552-6

Funding

Agricultural Science and Technology Independent Innovation Funding Program of Jiangsu Province(CX(24)3086)

Shandong Provincial Technology Innovation Guidance Program(YDZX2025104)

Jiangsu Basic Research Center for Synthetic Biology(BK20233003)

RIGHTS & PERMISSIONS

Jiangnan University