skip to main content

Preparation and Candida rugosa Lipase Immobilization on Nylon-6 Grafted and Aminated (Polyvinyl Benzyl Chloride) Microfibers

1Department of Biotechnology Engineering, International Islamic University Malaysia, 53100, Gombak, Malaysia

2International Institute for Halal Research & Training (INHART), International Islamic University Malaysia, 53100, Gombak, Malaysia

3Department of Chemical Engineering, Universiti Teknologi Petronas, 32610 Seri Iskandar, Perak, Malaysia

Received: 6 Jul 2018; Revised: 30 Jan 2019; Accepted: 2 Feb 2019; Available online: 30 Apr 2019; Published: 1 Aug 2019.
Editor(s): Istadi Istadi
Open Access Copyright (c) 2019 by Authors, Published by BCREC Group under http://creativecommons.org/licenses/by-sa/4.0.

Citation Format:
Cover Image
Abstract

This paper demonstrates a simplified procedure for the preparation of a nylon-6 microfibers based support for the immobilization of Candida rugosa lipase via covalent attachment to enhance the stability and reusability of lipase. The preparation of the support was done by radiation induced graft copolymerization (RIGC) of vinyl benzyl chloride (VBC) onto nylon-6 microfibers followed by amination with ethanolamine to facilitate the immobilization of lipase. Fourier transfer infra red (FTIR) and scanning electron microscope (SEM) were used to study the chemical and physical changes following grafting, amination and immobilization. Response surface methodology (RSM) was applied for the optimization of lipase immobilization on the aminated microfibers. The optimization parameters were incubation time, pH, and lipase concentration. Moreover, this study investigated the effect of temperature, pH, and storage stability and reusability on the lipase in its immobilized and free forms. The developed model from RSM showed an R2 value of 0.9823 and P-value < 0.001 indicating that the model is significant. The optimum temperatures for both immobilized and free lipases were 45 °C, whereas the best pH values for lipase activity were at pH 8 and pH 7, respectively. This study also identifies values for KM and Vmax for both immobilized and free lipase accordingly. Based on the results, immobilized lipase had significantly improved the stability and reusability of lipase compared to that in free forms. 

Fulltext View|Download
Keywords: Lipase immobilization; PVBC-grafted nylon-6 microfiber; amination; response surface methodology; optimization; enzyme activity
Funding: research grant (FRGS 14-162-0403) obtained under the Fundamental Research Grants Scheme, Ministry of Higher Education (MOHE), Malaysia

Article Metrics:

  1. Garcia-Galan, C., Berenguer-Murcia, Á., Fernandez-Lafuente, R., Rodrigues, R.C. (2011). Potential of Different Enzyme Immobilization Strategies to Improve Enzyme Performance. Adv. Synth. Catal., 353: 2885–2904
  2. Jegannathan, K.R., Abang, S., Poncelet, D., Chan, E.S., Ravindra, P. (2008). Production of biodiesel using immobilized lipase--a critical review. Crit. Rev. Biotechnol., 28: 253–264
  3. Miranda, M., Urioste, D., Andrade Souza, L.T., Mendes, A.A., de Castro, H.F. (2011), Assessment of the Morphological, Biochemical, and Kinetic Properties for Candida rugosa Lipase Immobilized on Hydrous Niobium Oxide to Be Used in the Biodiesel Synthesis. Enzyme Res., 2011: 1–10
  4. Kuo, C., Peng, L., Kan, S., Liu, Y., Shieh, C. (2013). Lipase-immobilized biocatalytic membranes for biodiesel production. Bioresou. Technol. 145: 229-232. doi: 10.1016/j.biortech.2012.12.054
  5. Brena, B., Batista-Viera, F. (2013). Immobilization of enzymes: A literature survey. Methods Mol. Biol., 1051: 15–31
  6. Narwal, S.K., Saun, N.K., Dogra, P., Chauhan, G., Gupta, R. (2015). Production and Characterization of Biodiesel Using Nonedible Castor Oil by Immobilized Lipase from Bacillus aerius. Biomed. Res. Int., 2015: 1: 1-6
  7. Xu, Z., Wan, L., Huang, X. (2009). Membrane with Biocatalytic Surface. Adv. Top. Sci. Technol. China, Springer. pp.263–305
  8. Isgrove, F.H., Williams, R.J.H., Niven, G.W., Andrews, A.T. (2001). Enzyme immobilization on nylon – optimization and the steps used to prevent enzyme leakage from the support. Enzym. Microbial. Technol. 28: 225–232
  9. Shalaby, S., Al-Balakoey, N., Abo El-Ola, S. (2007). Surface Modification of Nylon-6 Fibres for Medical Applications. Appl. Polym. Sci., 104: 3788–3796
  10. Shalaby, S., Al-Balakoey, N., Abdel-Fatah, O., Elshafei, A. (2008). Antimicrobial Finishing of Regular and Modified Nylon-6 Fabrics. Appl. Polym. Sci., 110(2): 738–746
  11. Shalaby, S.E., Beliakova, M.K. (2015). Simple Method for Grafting of Glycidyl Methacrylate onto Nylon-6 Nonwoven Fabrics for Ion Exchange Applications. Int. J. Sci. Res., 4: 689–696
  12. Liu W, Zhang S, Yu Jl, Chen X, Li L, Feng Q, Zhu X (2011) Surface Photografting: New Application for Flame Retardant Finishing of Polyamide6.6 (PA6.6) Fabric. Appl. Polym. Sci., 119: 66–72
  13. Li, X., Gu, X., Zhang, S., Li, H., Feng, Q., Sun, J., Zhao, Q. (2013). Improving the fire performance of nylon 6,6 fabric by chemical grafting with acrylamide. Ind. Eng. Chem. Res., 52: 2290–2296
  14. Nasef, M.M., Hegazy, E.S.A. (2004). Preparation and applications of ion exchange membranes by radiation-induced graft copolymerization of polar monomers onto non-polar films. Prog. Polym. Sci., 29: 499–561
  15. Ting, T.M., Nasef, M.M., Hashim, K. (2015). Modification of nylon-6 fibres by radiation-induced graft polymerisation of vinylbenzyl chloride. Radiat. Phys. Chem., 109: 54–62
  16. Ting, T.M., Nasef, M.M., Sithambaranathan, P. (2017). Kinetic investigations of emulsion- and solvent-mediated radiation induced graft copolymerization of glycidyl methacrylate onto nylon-6 fibres. J. Radioanal. Nucl. Chem., 311: 843–857
  17. Ikeda, K., Umeno, D., Saito, K., Koide, F., Miyata, E., Sugo, T. (2011). Removal of boron using nylon-based chelating fibers. Ind. Eng. Chem. Res., 50: 5727–5732
  18. Zdarta, J., Meyer, A., Jesionowski, T., Pinelo, M. (2018). A General Overview of Support Materials for Enzyme Immobilization: Characteristics, Properties, Practical Utility. Catalysts, 8: 92
  19. Nasef, M.M., Abbasi, A., Ting, T.M. (2014). New CO2 adsorbent containing aminated poly(glycidyl methacrylate) grafted onto irradiated PE-PP nonwoven sheet. Radiat. Phys. Chem., 103: 72–74
  20. Huang, X.J., Chen, P.C., Huang, F., Ou, Y., Chen, M.R., Xu, Z.K. (2011). Immobilization of Candida rugosa lipase on electrospun cellulose nanofiber membrane. J. Mol. Catal. B Enzym., 70: 95–100
  21. Bradford, M.M. (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
  22. Chiou, S.H., Wu, W.T. (2004). Immobilization of Candida rugosa lipase on chitosan with activation of the hydroxyl groups. Biomaterials, 25: 197–204
  23. Pahujani, S., Kanwar, S.S., Chauhan, G., Gupta, R. (2008). Glutaraldehyde activation of polymer Nylon-6 for lipase immobilization: Enzyme characteristics and stability. Bioresour. Technol., 99: 2566–2570
  24. Doǧaç, Y.I., Teke, M. (2013). Immobilization of bovine catalase onto magnetic nanoparticles. Prep. Biochem. Biotechnol., 43: 750–765
  25. Li, G., Nandgaonkar, A.G., Lu, K., Krause, W.E., Lucia, L.A., 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
  26. Osho, M., Popoola, T., Adeleye, T., Adetunji, C. (2016). Response surface methodology for optimal immobilization of Aspergillus niger ATCC 1015 lipase by adsorption method. Int. J. Biol. Res., 4: 56
  27. Zhu, J., Zhang, J., Lai, Q., Jiang, B., Gong, R. (2013). Covalent immobilization of lipase onto citric acid-esterified loofah sponge. BioResources, 8: 3289–3298
  28. Ibrahim, A.S.S., Al-Salamah, A.A., El-Toni, A.M., Almaary, K.S., El-Tayeb, M.A., Elbadawi, Y.B., Antranikian, G. (2016). Enhancement of alkaline protease activity and stability via covalent immobilization onto hollow core-mesoporous shell silica nanospheres. Int. J. Mol. Sci., 17(2): 184
  29. Su, R., Shi, P., Zhu, M., Hong, F., Li, D. (2012). Studies on the properties of graphene oxide-alkaline protease bio-composites. Bioresour. Technol., 115: 136–140
  30. Zhu, W., Zhang, Y., Hou, C., Pan, D., He, J., Zhu, H. (2016). Covalent immobilization of lipases on monodisperse magnetic microspheres modified with PAMAM-dendrimer. J. Nanoparticle Res., 18: 1–13
  31. Arıca, M.Y., Kaya, B., Bayramoglu, G. (2005). Food Chemistry Immobilization of Candida rugosa lipase onto spacer-arm attached poly (GMA-HEMA-EGDMA) microspheres. Food Chem., 92: 261–268
  32. Xu, D.Y., Yang, Y., Yang, Z. (2011). Activity and stability of cross-linked tyrosinase aggregates in aqueous and nonaqueous media. J. Biotechnol., 152: 30–36

Last update:

No citation recorded.

Last update:

No citation recorded.