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Synthesis of a polydopamaine nanoparticle/bacterial cellulose composite for use as a biocompatible matrix for laccase immobilization

  • Rui Zhai
  • Xiangxue Chen
  • Mingjie JinEmail author
  • Jinguang HuEmail author
Original Research


Bacterial cellulose has attracted attention as a scaffolding material for enzyme immobilization because of its excellent mechanical properties, nanoporous structure, high purity, and super hydrophilicity. However, the lack of reactive group and the relatively low binding capacity toward biomolecules limit its application. In this study, we have synthesized a polydopamine nanoparticle/bacterial cellulose hybrid material with hierarchical multiscale architecture for enzyme immobilization, via a simple in situ self-assembly of polydopamine nanoparticles on cellulose surface. Commercial enzyme laccases were employed to evaluate the enzyme immobilizability of this hybrid material, and various key parameters of enzyme immobilization such as reaction time, enzyme concentration, temperature and pH were also systematically studied. This hybrid composite displayed excellent enzyme immobilization efficiency (~ 165 mg laccase per g composite), and the immobilized laccase also exhibited better thermostability and operational stability over a broad temperature range as compared with free laccase. By using the immobilized laccase, fast and efficient dye removal ability and good recyclability were achieved during the dye decolorization test, which demonstrated its potential application in various industrial relevant wastewater treatment processes.

Graphic abstract


Bacterial cellulose Polydopamine nanoparticles Laccase Enzyme immobilization Wastewater treatment 



This work was financially supported by National Key R&D Program of China (Grant No. 2016YFE0105400), Natural Science and Engineering Research Council (NSERC) Canada First Research Excellence Fund, National Natural Science Foundation of China (Grant No. 21606132), Natural Science Foundation of Jiangsu Province (Grant Nos. BK20160823, BK20170037 and BK20170832), the Fundamental Research Funds for the Central Universities (Grant No. 30916011202), the Foundation of Jiangsu Specially-Appointed Professor and the Foundation of Jiangsu Innovative and Entrepreneurial Doctors. The authors thank Prof. Qiang Zhang from Nanjing University of Science and Technology for his assistance in FT-IR measurement.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Ansari SA, Husain Q (2012) Potential applications of enzymes immobilized on/in nano materials: a review. Biotechnol Adv 30:512–523. CrossRefGoogle Scholar
  2. Arola S, Tammelin T, Setälä H, Tullila A, Linder MB (2012) Immobilization–stabilization of proteins on nanofibrillated cellulose derivatives and their bioactive film formation. Biomacromol 13:594–603. CrossRefGoogle Scholar
  3. Batul R, Yu A, Bhave M, Khaliq A (2018) Synthesis of polydopamine nanoparticles for drug delivery applications. Microsc Microanal 24:1758–1759. CrossRefGoogle Scholar
  4. Bayramoglu G, Yilmaz M, Yakup Arica M (2010) Preparation and characterization of epoxy-functionalized magnetic chitosan beads: laccase immobilized for degradation of reactive dyes. Bioprocess Biosyst Eng 33:439–448. CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. CrossRefGoogle Scholar
  6. Buschle-Diller G, Zeronian SH (1992) Enhancing the reactivity and strength of cotton fibers. J Appl Polym Sci 45:967–979CrossRefGoogle Scholar
  7. Chao C, Liu J, Wang J, Zhang Y, Zhang B, Zhang Y, Xiang X, Chen R (2013) Surface modification of halloysite nanotubes with dopamine for enzyme immobilization. ACS Appl Mater Interfaces 5:10559–10564CrossRefGoogle Scholar
  8. Chao C, Zhang B, Zhai R, Xiang X, Liu J, Chen R (2014) Natural nanotube-based biomimetic porous microspheres for significantly enhanced biomolecule immobilization. ACS Sustain Chem Eng 2:396–403. CrossRefGoogle Scholar
  9. Chen L, Zou M, Hong FF (2015) Evaluation of fungal laccase immobilized on natural nanostructured bacterial cellulose. Front Microbiol 6:1–10. Google Scholar
  10. Chen J, Leng J, Yang X, Liao L, Liu L, Xiao A (2017) Enhanced performance of magnetic graphene oxide-immobilized laccase and its application for the decolorization of dyes. Molecules. Google Scholar
  11. Conte MP, Lau KHA, Ulijn RV (2017) Biocatalytic self-assembly using reversible and irreversible enzyme immobilization. ACS Appl Mater Interfaces 9:3266–3271. CrossRefGoogle Scholar
  12. Cristóvão RO, Tavares APM, Brígida AI, Loureiro JM, Boaventura RAR, Macedo EA, Coelho MAZ (2011) Immobilization of commercial laccase onto green coconut fiber by adsorption and its application for reactive textile dyes degradation. J Mol Catal B Enzym 72:6–12. CrossRefGoogle Scholar
  13. Drozd R, Rakoczy R, Wasak A, Junka A, Fijałkowski K (2018) The application of magnetically modified bacterial cellulose for immobilization of laccase. Int J Biol Macromol 108:462–470. CrossRefGoogle Scholar
  14. Frazão CJR, Silva NHC, Freire CSR, Silvestre AJD, Xavier AMRB, Tavares APM (2014) Bacterial cellulose as carrier for immobilization of laccase: optimization and characterization. Eng Life Sci 14:500–508. CrossRefGoogle Scholar
  15. Gholizadeh R, Wang Y (2018) Molecular dynamics simulation of the aggregation phenomenon in the late stages of silica materials preparation. Chem Eng Sci 184:62–71. CrossRefGoogle Scholar
  16. Gu GE, Park CS, Cho HJ, Ha TH, Bae J, Kwon OS, Lee JS, Lee CS (2018) Fluorescent polydopamine nanoparticles as a probe for zebrafish sensory hair cells targeted in vivo imaging. Sci Rep 8:1–8. CrossRefGoogle Scholar
  17. Hu W, Chen S, Yang J, Li Z, Wang H (2014) Functionalized bacterial cellulose derivatives and nanocomposites. Carbohydr Polym 101:1043–1060. CrossRefGoogle Scholar
  18. Huang Y, Li Y, Hu Z, Yue X, Proetto MT, Jones Y, Gianneschi NC (2017) Mimicking melanosomes: polydopamine nanoparticles as artificial microparasols. ACS Cent Sci 3:564–569. CrossRefGoogle Scholar
  19. Jia F, Narasimhan B, Mallapragada S (2014) Materials-based strategies for multi-enzyme immobilization and co-localization: a review. Biotechnol Bioeng 111:209–222. CrossRefGoogle Scholar
  20. Kadam AA, Jang J, Jee SC, Sung JS, Lee DS (2018) Chitosan-functionalized supermagnetic halloysite nanotubes for covalent laccase immobilization. Carbohydr Polym 194:208–216. CrossRefGoogle Scholar
  21. Karagoz B, Bayramoglu G, Altintas B, Bicak N, Arica MY (2010) Poly(glycidyl methacrylate)-polystyrene diblocks copolymer grafted nanocomposite microspheres from surface-initiated atom transfer radical polymerization for lipase immobilization: application in flavor ester synthesis. Ind Eng Chem Res 49:9655–9665. CrossRefGoogle Scholar
  22. Lee H, Rho J, Messersmith PB (2009) Facile conjugation of biomolecu les onto surfaces via mussel adhesive protein inspired coatings. Adv Mater 21:431–434. CrossRefGoogle Scholar
  23. Li G, Nandgaonkar AG, Lu K, Krause WE, Lucia LA, Wei Q (2016) Laccase immobilized on PAN/O-MMT composite nanofibers support for substrate bioremediation: a de novo adsorption and biocatalytic synergy. RSC Adv 6:41420–41427. CrossRefGoogle Scholar
  24. Li Z, Yang Y, Wang Z, Zhang X, Chen Q, Qian X, Liu N, Wei Y, Ji Y (2017) Polydopamine nanoparticles doped in liquid crystal elastomers for producing dynamic 3D structures. J Mater Chem A 5:6740–6746. CrossRefGoogle Scholar
  25. Liebscher J, Mrówczyński R, Scheidt HA, Filip C, Haìdade ND, Turcu R, Bende A, Beck S (2013) Structure of polydopamine: a never-ending story? Langmuir 29:10539–10548. CrossRefGoogle Scholar
  26. Lin J, Fan L, Miao R, Le X, Chen S, Zhou X (2015) Enhancing catalytic performance of laccase via immobilization on chitosan/CeO2 microspheres. Int J Biol Macromol 78:1–8. CrossRefGoogle Scholar
  27. Liu F, He X, Zhang J, Chen H, Zhang H, Wang Z (2015) Controllable synthesis of polydopamine nanoparticles in microemulsions with pH-activatable properties for cancer detection and treatment. J Mater Chem B 3:6731–6739. CrossRefGoogle Scholar
  28. Lu S, Yu J, Cheng Y, Wang Q, Barras A, Xu W, Szunerits S, Cornu D, Boukherroub R (2017) Preparation of silver nanoparticles/polydopamine functionalized polyacrylonitrile fiber paper and its catalytic activity for the reduction 4-nitrophenol. Appl Surf Sci 411:163–169. CrossRefGoogle Scholar
  29. Mechichi T, Mhiri N, Sayadi S (2006) Remazol brilliant blue R decolourization by the laccase from Trametes trogii. Chemosphere 64:998–1005. CrossRefGoogle Scholar
  30. Miletić N, Nastasović A, Loos K (2012) Immobilization of biocatalysts for enzymatic polymerizations: possibilities, advantages, applications. Bioresour Technol 115:126–135. CrossRefGoogle Scholar
  31. Nata IF, Sureshkumar M, Lee CK (2011) One-pot preparation of amine-rich magnetite/bacterial cellulose nanocomposite and its application for arsenate removal. RSC Adv 1:625–631. CrossRefGoogle Scholar
  32. Neto SLM, Matheus DR, Machado KMG (2009) Influence of pH on the growth, laccase activity and RBBR decolorization by tropical basidiomycetes. Brazilian Arch Biol Technol 52:1075–1082. CrossRefGoogle Scholar
  33. Othman AM, González-Domínguez E, Sanromán Á, Correa-Duarte M, Moldes D (2016) Immobilization of laccase on functionalized multiwalled carbon nanotube membranes and application for dye decolorization. RSC Adv 6:114690–114697. CrossRefGoogle Scholar
  34. Perrot D, Croutxé-Barghorn C, Allonas X (2016) Towards mussel-like on-demand coatings: light-triggered polymerization of dopamine through a photoinduced pH jump. Polym Chem 7:2635–2638. CrossRefGoogle Scholar
  35. Rekuć A, Bryjak J, Szymańska K, Jarzebski AB (2009) Laccase immobilization on mesostructured cellular foams affords preparations with ultra high activity. Process Biochem 44:191–198. CrossRefGoogle Scholar
  36. Rodríguez Couto S, Toca Herrera JL (2006) Industrial and biotechnological applications of laccases: a review. Biotechnol Adv 24:500–513. CrossRefGoogle Scholar
  37. Römling U, Galperin MY (2015) Bacterial cellulose biosynthesis: diversity of operons, subunits, products, and functions. Trends Microbiol 23:545–557. CrossRefGoogle Scholar
  38. Sampaio LMP, Padrão J, Faria J, Silva JP, Silva CJ, Dourado F, Zille A (2016) Laccase immobilization on bacterial nanocellulose membranes: antimicrobial, kinetic and stability properties. Carbohydr Polym 145:1–12. CrossRefGoogle Scholar
  39. Sulaiman S, Mokhtar MN, Naim MN, Baharuddin AS, Sulaiman A (2014) A review: potential usage of cellulose nanofibers (CNF) for enzyme immobilization via covalent interactions. Appl Biochem Biotechnol 175:1817–1842. CrossRefGoogle Scholar
  40. Suwannawong P, Khammuang S, Sarnthima R (2010) Decolorization of rhodamine B and congo red by partial purified laccase from Lentinus polychrous Lév. J Biochem Technol 2:182–186Google Scholar
  41. Taheran M, Naghdi M, Brar SK, Knystautas EJ, Verma M, Surampalli RY (2017) Covalent immobilization of laccase onto nanofibrous membrane for degradation of pharmaceutical residues in water. ACS Sustain Chem Eng 5:10430–10438. CrossRefGoogle Scholar
  42. Tavares APM, Silva CG, Dražić G, Silva AMT, Loureiro JM, Faria JL (2015) Laccase immobilization over multi-walled carbon nanotubes: kinetic, thermodynamic and stability studies. J Colloid Interface Sci 454:52–60. CrossRefGoogle Scholar
  43. Ullah H, Santos HA, Khan T (2016) Applications of bacterial cellulose in food, cosmetics and drug delivery. Cellulose 23:2291–2314. CrossRefGoogle Scholar
  44. Wesenberg D, Kyriakides I, Agathos SN (2003) White-rot fungi and their enzymes for the treatment of industrial dye effluents. Biotechnol Adv 22:161–187. CrossRefGoogle Scholar
  45. Xia TT, Liu CZ, Hu JH, Guo C (2016) Improved performance of immobilized laccase on amine-functioned magnetic Fe3O4 nanoparticles modified with polyethylenimine. Chem Eng J 295:201–206. CrossRefGoogle Scholar
  46. Yeroslavsky G, Girshevitz O, Foster-Frey J, Donovan DM, Rahimipour S (2015) Antibacterial and antibiofilm surfaces through polydopamine-assisted immobilization of lysostaphin as an antibacterial enzyme. Langmuir 31:1064–1073. CrossRefGoogle Scholar
  47. Zhai R, Zhang B, Wan Y, Li C, Wang J, Liu J (2013) Chitosan-halloysite hybrid-nanotubes: horseradish peroxidase immobilization and applications in phenol removal. Chem Eng J 214:304–309. CrossRefGoogle Scholar
  48. Zhang J, Song M, Wang X, Wu J, Yang Z, Cao J, Chen Y, Wei Q (2016) Preparation of a cellulose acetate/organic montmorillonite composite porous ultrafine fiber membrane for enzyme immobilization. J Appl Polym Sci 133:1–8. Google Scholar
  49. Zhang H, Luo J, Li S, Wei Y, Wan Y (2018) Biocatalytic membrane based on polydopamine coating: a platform for studying immobilization mechanisms. Langmuir 34:2585–2594. CrossRefGoogle Scholar
  50. Zhu Z, Su M (2017) Polydopamine nanoparticles for combined chemo- and photothermal cancer therapy. Nanomaterials 7:160. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.School of Environmental and Biological EngineeringNanjing University of Science and TechnologyNanjingChina
  2. 2.Department of Chemical and Petroleum EngineeringUniversity of CalgaryCalgaryCanada

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