One-Pot Lipase Entrapment Within Silica Particles to Prepare a Stable and Reusable Biocatalyst for Transesterification

Original Paper

Abstract

In order to enhance the reusability, Rhizomucor miehei lipase was entrapped in a single step within silica particles having an oleic acid core (RML@SiO2). Characterization of RML@SiO2 by scanning and transmission electron microscopy and Fourier transform infrared studies supported the lipase immobilization within silica particles. The immobilized enzyme was employed for transesterification of cottonseed oil with methanol and ethanol. Under the optimum reaction conditions of a methanol-to-oil molar ratio of 12:1 or ethanol-to-oil molar ratio of 15:1, stirring speed of 250 revolutions/min (flask radius = 3 cm), reaction temperature of 40 °C, and biocatalyst concentration of 5 wt% (with respect to oil), more than 98 % alkyl ester yield was achieved in 16 and 24 h of reaction duration in case of methanolysis and ethanolysis, respectively. The immobilized enzyme did not require any buffer solution or organic solvent for optimum activity; hence, the produced biodiesel and glycerol were free from metal ion or organic molecule contamination. The activation energies for the immobilized enzyme-catalyzed ethanolysis and methanolysis were found to be 34.9 ± 1.6 and 19.7 ± 1.8 kJ mol−1, respectively. The immobilized enzyme was recovered from the reaction mixture and reused in 12 successive runs without significant loss of activity. Additionally, RML@SiO2 demonstrated better reusability as well as stability in comparison to the native enzyme as the former did not lose the activity even upon storage at room temperature (25–30 °C) over an 8-month period.

Keywords

Immobilization Rhizomucor miehei lipase Kinetics Enzyme stability Biodiesel 

Supplementary material

11746_2015_2630_MOESM1_ESM.docx (56 kb)
Supplementary material 1 (DOCX 55 kb)

References

  1. 1.
    Al-Zuhair S, Almenhali A, Hamad I, Alshehhi M, Alsuwaidi N, Mohamed S (2011) Enzymatic production of biodiesel from used/waste vegetable oils: design of a pilot plant. Renew Energ 36:2605–2614CrossRefGoogle Scholar
  2. 2.
    Madankar CS, Pradhana S, Naik SN (2013) Parametric study of reactive extraction of castor seed (Ricinus communis L.) for methyl ester production and its potential use as bio lubricant. Ind Crop Prod 43:283–290CrossRefGoogle Scholar
  3. 3.
    Pradhan S, Madankar CS, Mohanty P, Naik SN (2012) Optimization of reactive extraction of castor seed to produce biodiesel using response surface methodology. Fuel 97:848–855CrossRefGoogle Scholar
  4. 4.
    Al-Zuhair S, Ling FW, Jun LS (2007) Proposed kinetic mechanism of the production of biodiesel from palm oil using lipase. Process Biochem 42:951–960CrossRefGoogle Scholar
  5. 5.
    Karimpil JJ, Melo JS, D’Souza SF (2012) Immobilization of lipase on cotton cloth using the layer-by-layer self-assembly technique. Int J Biol Macromol 50:300–302CrossRefGoogle Scholar
  6. 6.
    Katiyar M, Ali A (2012) Immobilization of Candida rugosa lipase on MCM-41 for the transesterification of cotton seed oil. J Oleo Sci 61:469–475CrossRefGoogle Scholar
  7. 7.
    Karimpil JJ, Melo JS, D’Souza SF (2011) Hen egg white as a feeder protein for lipase immobilization. J Mol Catal B Enzym 71:113–118CrossRefGoogle Scholar
  8. 8.
    Tan T, Lu J, Nie K, Deng L, Wang F (2010) Biodiesel production with immobilized lipase: a review. Biotechnol Adv 28:628–634CrossRefGoogle Scholar
  9. 9.
    Souza RL, Resende WC, Barao CE, Zanin GM, De-Castro HF, Santos OAA, Fricks AT, Figueiredo RT, Lima AS, Soares CMF (2012) Influence of the use of Aliquat 336 in the immobilization procedure in sol–gel of lipase from Bacillus sp. ITP-001. J Mol Catal B Enzym 84:152–159CrossRefGoogle Scholar
  10. 10.
    Hartmann M, Jung D (2010) Biocatalysis with enzymes immobilized on mesoporous hosts: the status quo and future trends. J Mater Chem 20:844–857CrossRefGoogle Scholar
  11. 11.
    Kuwahara Y, Yamanishi T, Kamegawa T, Mori K, Che M, Yamashita H (2012) Lipase-embedded silica nanoparticles with oil-filled core–shell structure: stable and recyclable platforms for biocatalysts. Chem Commun 48:2882–2884CrossRefGoogle Scholar
  12. 12.
    Knothe G (2001) Determining the blend level of mixtures of biodiesel with conventional diesel fuel by fiber-optic near-infrared spectroscopy and 1H nuclear magnetic resonance spectroscopy. J Am Oil Chem Soc 78:1025–1028CrossRefGoogle Scholar
  13. 13.
    Ghesti GF, De-Macedo JL, Resck IS, Dias JA, Dias SCL (2007) FT-Raman spectroscopy quantification of biodiesel in a progressive soybean oil transesterification reaction and its correlation with 1H NMR spectroscopy methods. Energy Fuel 21:2475–2480CrossRefGoogle Scholar
  14. 14.
    Zhou Z, Hartmann M (2012) Recent progress in biocatalysis with enzymes immobilized on mesoporous hosts. Top Catal 55:1081–1100CrossRefGoogle Scholar
  15. 15.
    Bai G, Zhou G, Li Y, Li T, Zhang X (2010) Synthesis of mesoporous silica templated by Pluronic F68 and its application in the immobilization of lipase. J Porous Mat 17:755–761CrossRefGoogle Scholar
  16. 16.
    Huang D, Han S, Han Z, Lin Y (2012) Biodiesel production catalyzed by Rhizomucor miehei lipase-displaying Pichia pastoris whole cells in an isooctane system. Biochem Eng J 63:10–14CrossRefGoogle Scholar
  17. 17.
    Ranganathan SV, Narasimhan SL, Muthukumar K (2008) An overview of enzymatic production of biodiesel. Bioresour Technol 99:3975–3981CrossRefGoogle Scholar
  18. 18.
    Meunier SM, Legge RL (2012) Evaluation of diatomaceous earth supported lipase sol–gels as a medium for enzymatic transesterification of biodiesel. J Mol Catal B Enzym 77:92–97CrossRefGoogle Scholar
  19. 19.
    Kumar D, Ali A (2012) Nanocrystalline K-CaO for the transesterification of a variety of feedstocks: structure, kinetics and catalytic properties. Biomass Bioenerg 46:459–468CrossRefGoogle Scholar
  20. 20.
    Song R, Tong D, Tang J, Hu C (2011) Effect of composition on the structure and catalytic properties of KF/Mg–La solid base catalysts for biodiesel synthesis via transesterification of cottonseed oil. Energy Fuel 25:2679–2686CrossRefGoogle Scholar
  21. 21.
    Pogaku R, Raman JK, Kumar GR (2012) Evaluation of activation energy and thermodynamic properties of enzyme-catalysed transesterification reactions. Adv Chem Eng Sci 1:150–154CrossRefGoogle Scholar

Copyright information

© AOCS 2015

Authors and Affiliations

  1. 1.School of Chemistry and BiochemistryThapar UniversityPatialaIndia

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