Immobilized enzyme on pulp fiber through layer-by-layer technique using cationic polyacrylamide for whitewater treatment from papermaking

  • Rina WuEmail author
  • Qiuyu Wang
  • Gaosheng Wang
Research Paper


Anionic pectic substances in whitewater from papermaking are detrimental to machine operation and product quality. Pectinase was immobilized on pulp fiber using cationic polyacrylamide with layer-by-layer method to obtain bound enzyme with tunable activity and good performance for wastewater treatment. It was revealed that high charge density and low molecular weight for cationic polyacrylamide were advantageous for enzymatic activity. During the layer-by-layer adsorption process, the enzymatic activity of the immobilized enzyme increased nearly linearly with the layer number from 983 to 3074 U/g until the fourth layer. The stability of the four-layer immobilized enzyme was improved. The multilayer immobilized enzyme exhibited good reusability and storage stability compared with monolayer enzyme. At dosage of 10 U/mL, the cationic demand of the whitewater samples was reduced by 15% using four-layer immobilized enzyme. The results indicated a potential route to prepare immobilized enzyme with good performance for wastewater treatment in papermaking industry.


Enzyme immobilization Pulp fiber Layer-by-layer Papermaking Cationic polyacrylamide 



This work was supported by State Key Laboratory of Pulp and Paper Engineering (201502), Innovation fund for young teachers of Tianjin University of Science and Technology (2016LG19), and Key project of Tianjin Natural Science Foundation (16JCZDJC39700).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Choi J, Han S, Kim H (2015) Industrial applications of enzyme biocatalysis: current status and future aspects. Biotechnol Adv 33:1443–1454CrossRefGoogle Scholar
  2. 2.
    Tsatsis DE, Papachristos DK, Valta KA, Vlyssides AG, Economides DG (2017) Enzymatic deinking for recycling of office waste paper. J Environ Chem Eng 5:1744–1753CrossRefGoogle Scholar
  3. 3.
    Aracri E, Colom JF, Vidal T (2009) Application of laccase-natural mediator systems to sisal pulp: An effective approach to biobleaching or functionalizing pulp fibres? Bioresour Technol 100:5911–5916CrossRefGoogle Scholar
  4. 4.
    Ramírez-Tapias YA, Rivero CW, Britos CN, Trelles JA (2015) Alkaline and thermostable polygalacturonase from Streptomyces halstedii ATCC 10897 with applications in waste waters. Biocatal Agric Biotechnol 4:221–228CrossRefGoogle Scholar
  5. 5.
    Li Z, Qin Y, Qin M, Liu N, Xu Q, Fu Y, Yuan Z (2012) Changes of anionic groups in alkaline peroxide-impregnated aspen chemithermomechanical pulp during subsequent alkaline peroxide bleaching. Carbohyd Polym 88:1041–1046CrossRefGoogle Scholar
  6. 6.
    Ricard M, Reid I, Orccotoma JA (2005) Pectinase reduces the cationic demand of peroxide-bleached TMP: a paper machine trial. Pulp Pap Can 106:78–83Google Scholar
  7. 7.
    Liu K, Zhao G, He B, Chen L, Huang L (2012) Immobilization of pectinase and lipase on macroporous resin coated with chitosan for treatment of whitewater from papermaking. Bioresour Technol 123:616–619CrossRefGoogle Scholar
  8. 8.
    Barbosa O, Ortiz C, Berenguer-Murcia Á, Torres R, Rodrigues RC, Fernandez-Lafuente R (2015) Strategies for the one-step immobilization–purification of enzymes as industrial biocatalysts. Biotechnol Adv 33:435–456CrossRefGoogle Scholar
  9. 9.
    Liu D, Chen J, Shi Y (2018) Advances on methods and easy separated support materials for enzymes immobilization. TrAC Trends Anal Chem 102:332–342CrossRefGoogle Scholar
  10. 10.
    Lisdat F (2017) Trends in the layer-by-layer assembly of redox proteins and enzymes in bioelectrochemistry. Curr Opin Electrochem 5:165–172CrossRefGoogle Scholar
  11. 11.
    Raghuwanshi VS, Garusinghe UM, Raj P, Kirby N, Hoell A, Batchelor W, Garnier G (2018) Cationic polyacrylamide induced nanoparticles assembly in a cellulose nanofiber network. J Colloid Interface Sci 529:180–186CrossRefGoogle Scholar
  12. 12.
    Wang Y, Chen K, Mo L, Li J, Xu J (2014) Optimization of coagulation–flocculation process for papermaking-reconstituted tobacco slice wastewater treatment using response surface methodology. J Ind Eng Chem 20:391–396CrossRefGoogle Scholar
  13. 13.
    Rehman HU, Aman A, Nawaz MA, Karim A, Ghani M, Baloch AH, Qader SAU (2016) Immobilization of pectin depolymerising polygalacturonase using different polymers. Int J Biol Macromol 82:127–133CrossRefGoogle Scholar
  14. 14.
    Wang Q, Liu S, Yang G, Chen J, Ni Y (2015) Cationic polyacrylamide enhancing cellulase treatment efficiency of hardwood kraft-based dissolving pulp. Bioresour Technol 183:42–46CrossRefGoogle Scholar
  15. 15.
    John M (2002) The protein protocols handbook. Humana Press, New JerseyGoogle Scholar
  16. 16.
    Wu R, He B, Zhao G, Qian L, Li X (2013) Immobilization of pectinase on oxidized pulp fiber and its application in whitewater treatment. Carbohyd Polym 97:523–529CrossRefGoogle Scholar
  17. 17.
    Wågberg L, Hägglund R (2001) Kinetics of polyelectrolyte adsorption on cellulosic fibers. Langmuir 17:1096–1103CrossRefGoogle Scholar
  18. 18.
    Guzmán E, Mateos-Maroto A, Ruano M, Ortega F, Rubio RG (2017) Layer-by-Layer polyelectrolyte assemblies for encapsulation and release of active compounds. Adv Colloid Interface 249:290–307CrossRefGoogle Scholar
  19. 19.
    Xing Q, Eadula SR, Lvov YM (2007) Cellulose fiber-enzyme composites fabricated through layer-by-layer nanoassembly. Biomacromolecules 8:1987–1991CrossRefGoogle Scholar
  20. 20.
    And FC, Schüler C (2000) Enzyme multilayers on colloid particles: assembly, stability, and enzymatic activity. Langmuir 16:9595–9603CrossRefGoogle Scholar
  21. 21.
    Fernandez-Lopez L, Pedrero SG, Lopez-Carrobles N, Gorines BC, Virgen-Ortíz JJ, Fernandez-Lafuente R (2017) Effect of protein load on stability of immobilized enzymes. Enzyme Microb Technol 98:18–25CrossRefGoogle Scholar
  22. 22.
    Abdel-Naby MA (1999) Immobilization of Paenibacillus macerans NRRL B-3186 cyclodextrin glucosyltransferase and properties of the immobilized enzyme. Process Biochem 34:399–405CrossRefGoogle Scholar
  23. 23.
    Guedidi S, Yurekli Y, Deratani A, Déjardin P, Innocent C, Altinkaya SA, Roudesli S, Yemenicioglu A (2010) Effect of enzyme location on activity and stability of trypsin and urease immobilized on porous membranes by using layer-by-layer self-assembly of polyelectrolyte. J Membr Sci 365:59–67CrossRefGoogle Scholar
  24. 24.
    Talbert JN, Goddard JM (2012) Enzymes on material surfaces. Colloids Surf B 93:8–19CrossRefGoogle Scholar
  25. 25.
    Lei Z, Jiang Q (2011) Synthesis and properties of immobilized pectinase onto the macroporous polyacrylamide microspheres. J Agric Food Chem 59:2592–2599CrossRefGoogle Scholar
  26. 26.
    Pečar D, Goršek A (2017) Process and kinetic characteristics of glucose oxidation catalyzed with immobilized enzyme. React Kinet Mech Catal 122:43–51CrossRefGoogle Scholar
  27. 27.
    Ferreira MM, Santiago FLB, Silva NAGD, Luiz JHH, Fernandéz-Lafuente R, Mendes AA, Hirata DB (2018) Different strategies to immobilize lipase from Geotrichum candidum: kinetic and thermodynamic studies. Process Biochem 67:55–63CrossRefGoogle Scholar
  28. 28.
    de Oliveira RL, Dias JL, Da Silva OS, Porto TS (2018) Immobilization of pectinase from Aspergillus aculeatus in alginate beads and clarification of apple and umbu juices in a packed bed reactor. Food Bioprod Process 109:9–18CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Tianjin Key Laboratory of Pulp and PaperTianjin University of Science and TechnologyTianjinChina
  2. 2.State Key Laboratory of Pulp and Paper EngineeringSouth China University of TechnologyGuangzhouChina

Personalised recommendations