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Bioprocess and Biosystems Engineering

, Volume 41, Issue 7, pp 991–1002 | Cite as

Kinetic and thermodynamic studies on the enzymatic synthesis of wax ester catalyzed by lipase immobilized on glutaraldehyde-activated rice husk particles

  • Letícia C. D. Lima
  • Daniela G. C. Peres
  • Adriano A. Mendes
Research Paper

Abstract

Commercial lipase from Thermomyces lanuginosus has been immobilized on glutaraldehyde-activated rice husk particles via covalent attachment. It was reached maximum immobilized protein concentration of 27.5 ± 1.8 mg g−1 of dry support using the initial protein loading of 40 mg g−1 of support. The immobilized biocatalyst was used to synthesize cetyl oleate (wax ester) via direct esterification of oleic acid and cetyl alcohol. The influence of relevant factors on ester synthesis, such as reaction temperature, biocatalyst concentration, presence or lack of hydrophobic organic solvents, acid:alcohol molar ratio, and reaction time has been evaluated. The experimental data were well fitted to a second-order reversible kinetic model to determine apparent kinetic constants. Thermodynamic studies have revealed that the reaction was a spontaneous and endothermic process. Under optimal experimental conditions, it was observed maximum ester conversion of 90.2 ± 0.6% in 9 h of reaction time in hexane medium using 1 M of each reactant (cetyl alcohol and oleic acid), at 50 °C and biocatalyst concentration of 15% m/v of reaction mixture. Similar conversion (91.5 ± 0.8%) in a solvent-free system was also obtained within 24 h of reaction. The biocatalyst retained 85% of its initial activity after 12 cycles within 9 h of reaction in hexane medium. The physicochemical properties of purified ester have been determined in accordance with ASTM standards. The results indicate that the prepared biocatalyst has great potential for wax ester synthesis due to its satisfactory catalytic activity and operational stability.

Keywords

Rice husk Covalent attachment Lipase Wax ester synthesis Kinetic/thermodynamic studies 

Notes

Acknowledgements

This study was financially supported by FAPEMIG (Process APQ-02196-15), CNPq (Process 404929/2016-8) and CAPES (Brazil). Adriano A. Mendes thanks the CNPq Foundation for the research fellowship (PQ-2 CA EQ, Grant 301355/2017-7).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

References

  1. 1.
    Kuo CH, Chen HH, Chen JH, Liu YC, Shieh CJ (2012) High yield of wax ester synthesized from cetyl alcohol and octanoic acid by Lipozyme RMIM and Novozym 435. Int J Mol Sci 13:11694–11704CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Ungcharoenwiwat P, H-Kittikun A (2015) Purification and characterization of lipase from Burkholderia sp. EQ3 isolated from wastewater from a canned fish factory and its application for the synthesis of wax esters. J Mol Catal B Enzym 115:96–104CrossRefGoogle Scholar
  3. 3.
    Ungcharoenwiwat P, H-Kittikun A (2013) Synthesis of wax esters from crude fish fat by lipase of Burkholderia sp. EQ3 and commercial lipases. J Am Oil Chem Soc 90:359–367CrossRefGoogle Scholar
  4. 4.
    Li D, Xiaojing W, Kaili N, Fang W, Jufeng L, Pu W, Tianwei T (2011) Synthesis of wax esters by lipase-catalyzed esterification with immobilized lipase from Candida sp. 99–125. Chin J Chem Eng 19:978–982CrossRefGoogle Scholar
  5. 5.
    Rani KNP, Neeharika TSVR., Kumar TP, Satyavathi B, Sailu C, Prasad RBN (2015) Kinetics of enzymatic esterification of oleic acid and decanol for wax ester and evaluation of its physico-chemical properties. J Taiwan Inst Chem Eng 55:12–16CrossRefGoogle Scholar
  6. 6.
    Serrano-Arnaldos M, Máximo-Martín MF, Montiel-Morte MC, Ortega-Requena S, Gómez-Gómez E, Bastida-Rodríguez J (2016) Solvent-free enzymatic production of high quality cetyl esters. Bioprocess Biosyst Eng 39:641–649CrossRefPubMedGoogle Scholar
  7. 7.
    Alves MD, Cren EC, Mendes AA (2016) Kinetic, thermodynamic, optimization and reusability studies for the enzymatic synthesis of a saturated wax ester. J Mol Catal B Enzym 133:S377–S387CrossRefGoogle Scholar
  8. 8.
    Mendes AA, Castro HF, Rodrigues DS, Adriano WS, Tardioli PW, Mammarella EJ, Giordano RC, Giordano RLC (2011) Multipoint covalent immobilization of lipase on chitosan hybrid hydrogels: influence of the polyelectrolyte complex type and chemical modification on the catalytic properties of the biocatalysts. J Ind Microbiol Biotechnol 38:1055–1066CrossRefPubMedGoogle Scholar
  9. 9.
    Bezbradica DI, Mateo C, Guisan JM (2014) Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde. J Mol Catal B Enzym 102:218–224CrossRefGoogle Scholar
  10. 10.
    Badgujar VC, Badgujar KC, Yeole PM, Bhanage BM (2017) Immobilization of Rhizomucor miehei lipase on a polymeric film for synthesis of important fatty acid esters: kinetics and application studies. Bioprocess Biosyst Eng 40:1463–1478CrossRefPubMedGoogle Scholar
  11. 11.
    Lage FAP, Bassi JJ, Corradini MCC, Todero LM, Luiz JHH, Mendes AA (2016) Preparation of a biocatalyst via physical adsorption of lipase from Thermomyces lanuginosus on hydrophobic support to catalyze biolubricant synthesis by esterification reaction in a solvent-free system. Enzyme Microb Technol 84:56–67CrossRefPubMedGoogle Scholar
  12. 12.
    Costa-Silva TA, Cognette RC, Souza CRF, Said S, Oliveira WP (2013) Spouted bed drying as a method for enzyme immobilization. Drying Technol 31:1756–1763CrossRefGoogle Scholar
  13. 13.
    Costa-Silva TA, Carvalho AKF, Souza CRF, Castro HF, Said S, Oliveira WP (2016) Enzymatic synthesis of biodiesel using immobilized lipase on a non-commercial support. Energy Fuels 30:4820–4824CrossRefGoogle Scholar
  14. 14.
    Corici L. Ferrario V, Pellis A, Ebert C, Lotteria S, Cantone S, Voinovich D, Gardossi L (2016) Large scale applications of immobilized enzymes call for sustainable and inexpensive solutions: rice husks as renewable alternatives to fossil-based organic resins. RSC Adv 6:63256–63270CrossRefGoogle Scholar
  15. 15.
    He M, Li Y, Pi F, Ji J, He X, Zhang Y, Sun X (2016) A novel detoxifying agent: using rice husk carriers to imobilize zearalenone-degrading enzyme from Aspergillus niger FS10. Food Control 68:271–279CrossRefGoogle Scholar
  16. 16.
    Mateo C, Palomo JM, Fernandez-Lorente G, Guisan JM, Fernandez-Lafuente R (2007) Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme Microb Technol 40:1451–1463CrossRefGoogle Scholar
  17. 17.
    DiCosimo R, McAuliffe J, Poulose AJ, Bohlmann G (2013) Industrial use of immobilized enzymes. Chem Soc Rev 42:6437–6474CrossRefPubMedGoogle Scholar
  18. 18.
    Monsan P, Puzo G, Mazarguil H (1975) Étude du mécanisme d’établissement des liaisons glutaraldehyde protéines. Biochimie 57:1281–1292CrossRefPubMedGoogle Scholar
  19. 19.
    Monsan P (1978) Optimization of glutaraldehyde activation of a support for enzyme immobilization. J Mol Catal 3:371–384CrossRefGoogle Scholar
  20. 20.
    Migneault I, Dartiguenave C, Bertrand MJ, Waldron KC (2004) Glutaraldehyde: behavior in aqueous solution, reaction with proteins, and application to enzyme crosslinking. Biotechniques 37:790–802CrossRefPubMedGoogle Scholar
  21. 21.
    Silva JA, Macedo GP, Rodrigues DS, Giordano RLC, Gonçalves LRB (2012) Immobilization of Candida antarctica lipase B by covalent attachment on chitosan-based hydrogels using different support activation strategies. Biochem Eng J 60:16–24CrossRefGoogle Scholar
  22. 22.
    Barbosa O, Ortiz C, Berenguer-Murcia A, Torres R, Rodrigues RC, Fernandez-Lafuente R (2014) Glutaraldehyde in bio-catalysis design: a useful crosslinker and a versatile tool in enzyme immobilization. RSC Adv 4:1583–1600CrossRefGoogle Scholar
  23. 23.
    Zaak H, Peirce S, de Albuquerque TL, Sassi M, Fernandez-Lafuente R (2017) Exploiting the versatility of aminated supports activated with glutaraldehyde to immobilize β-galactosidase from Aspergillus oryzae. Catalysts 7:250CrossRefGoogle Scholar
  24. 24.
    Vazquez-Ortega PG, Alcaraz-Fructuoso MT, Rojas-Contreras JA, López-Miranda J, Fernandez-Lafuente R (2018) Stabilization of dimeric β-glucosidase from Aspergillus niger via glutaraldehyde immobilization under different conditions. Enzyme Microb Technol 110:38–45CrossRefPubMedGoogle Scholar
  25. 25.
    Zhang L, Yu P, Luo Y (2007) Comparative behavior of PVA/PAN and PVA/PES composite pervaporation membranes in the pervaporative dehydration of caprolactam. J Appl Polym Sci 103:4005–4011CrossRefGoogle Scholar
  26. 26.
    Wang Y, Hsieh YL (2010) Aldehyde functionalized cellulose support for hydrogels. J Appl Polym Sci 118:2489–2495CrossRefGoogle Scholar
  27. 27.
    Habeeb AFSA., Hiramoto R (1968) Reaction of proteins with glutaraldehyde. Arch Biochem Biophys 126:16–26CrossRefPubMedGoogle Scholar
  28. 28.
    Adriano WS, Mendonça DB, Rodrigues DS, Mammarella EJ, Giordano RLC (2008) Improving the properties of chitosan as support for the covalent multipoint immobilization of chymotrypsin. Biomacromol 9:2170–2179CrossRefGoogle Scholar
  29. 29.
    Manrich A, Galvão CMA, Jesus CDF, Giordano RC, Giordano RLC (2008) Immobilization of trypsin on chitosan gels: use of different activation protocols and comparison with other supports. Int J Biol Macromol 43:54–61CrossRefPubMedGoogle Scholar
  30. 30.
    Aissa M, Sellami M, Kamoun A, Gargouri Y, Miled N (2012) Optimization of immobilized lipase-catalyzed synthesis of wax esters by response surface methodology. Curr Chem Biol 6:77–85Google Scholar
  31. 31.
    Khan NR, Jadhav SV, Rathod VK (2015) Lipase catalysed synthesis of cetyl oleate using ultrasound: optimisation and kinetic studies. Ultrason Sonochem 27:522–529CrossRefPubMedGoogle Scholar
  32. 32.
    Virgen-Ortíz JJ, Tacias-Pascacio VG, Hirata DB, Torrestiana-Sanchez B, Rosales-Quintero A, Fernandez-Lafuente R (2017) Relevance of substrates and products on the desorption of lipases physically adsorbed on hydrophobic supports. Enzyme Microb Technol 96:30–35CrossRefPubMedGoogle Scholar
  33. 33.
    Virgen-Ortíz JJ, Pedrero SG, Fernandez-Lopez L, Lopez-Carrobles N, Gorines BC, Otero C, Fernandez-Lafuente R (2017) Desorption of lipases immobilized on octyl-agarose beads and coated with ionic polymers after termal inactivation. Stronger adsorption of polymers/unfolded protein composites. Molecules 22:91CrossRefGoogle Scholar
  34. 34.
    Vescovi V, Kopp W, Guisán JM, Giordano RLC, Mendes AA, Tardioli PW (2016) Improved catalytic properties of Candida antarctica lipase B multi-attached on tailor-made hydrophobic silica containing octyl and multifunctional amino-glutaraldehyde spacer arms. Process Biochem 51:2055–2066CrossRefGoogle Scholar
  35. 35.
    Albuquerque TL, Rueda N, Santos JCS, Barbosa O, Ortiz C, Binay B, Özdemir E, Gonçalves LRB, Fernandez-Lafuente R (2016) Easy stabilization of interfacially activated lipases using heterofunctional divinyl sulfone activated-octyl agarose beads. Modulation of the immobilized enzymes by altering their nanoenvironment. Process Biochem 51:865–874CrossRefGoogle Scholar
  36. 36.
    Hirata DB, Albuquerque TL, Rueda N, Virgen-Ortíz JJ, Tacias-Pascacio VG, Fernandez-Lafuente R (2016) Evaluation of different immobilized lipases in transesterification reactions using tributyrin: advantages of the heterofunctional octyl agarose beads. J Mol Catal B Enzym 133:117–123CrossRefGoogle Scholar
  37. 37.
    Fernandez-Lafuente R (2010) Lipase from Thermomyces lanuginosus: uses and prospects as an industrial biocatalyst. J Mol Catal B Enzym 62:197–212CrossRefGoogle Scholar
  38. 38.
    Bressani APP, Garcia KCA, Hirata DB, Mendes AA (2015) Production of alkyl esters from macaw palm oil by a sequential hydrolysis/esterification process using heterogeneous biocatalysts: optimization by response surface methodology. Bioprocess Biosyst Eng 38:287–297CrossRefPubMedGoogle Scholar
  39. 39.
    Todero LM, Bassi JJ, Lage FAP, Corradini MCC, Barboza JCS, Hirata DB, Mendes AA (2015) Enzymatic synthesis of isoamyl butyrate catalyzed by immobilized lipase on poly-methacrylate particles: optimization, reusability and mass transfer studies. Bioprocess Biosyst Eng 38:1601–1613CrossRefPubMedGoogle Scholar
  40. 40.
    Gupta A, Dhakate SR, Pahwa M, Sinha S, Chand S, Mathur RB (2013) Geranyl acetate synthesis catalyzed by Thermomyces lanuginosus lipase immobilized on electrospun polyacrylonitrile nanofiber membrane. Process Biochem 48:124–132CrossRefGoogle Scholar
  41. 41.
    Rueda N, Albuquerque TL, Bartolome-Cabrero R, Fernandez-Lopez L, Torres R, Ortiz C, Santos JCS, Barbosa O, Fernandez-Lafuente R (2016) Reversible immobilization of lipases on heterofunctional octyl-amino agarose beads prevents enzyme desorption. Molecules 21:646CrossRefGoogle Scholar
  42. 42.
    Vescovi V, Giordano RLC, Mendes AA, Tardioli PW (2017) Immobilized lipases on functionalized silica particles as potential biocatalysts for the synthesis of fructose oleate in an organic solvent/water system. Molecules 22:212CrossRefGoogle Scholar
  43. 43.
    Gupta MN, Roy I (2004) Enzymes in organic media – Forms, functions and applications. Eur J Biochem 271:2575–2583CrossRefPubMedGoogle Scholar
  44. 44.
    Sharma S, Kanwar SS (2014) Organic solvent tolerant lipases and applications. Sci World J 2014:625258Google Scholar
  45. 45.
    Kumar A, Dhar K, Kanwar SS, Arora PK (2016) Lipase catalysis in organic solvents: advantages and applications. Biol Proced Online 18:2CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Nuijens T, Cusan C, Schepers ACHM., Kruijtzer JAW, Rijkers DTS, Liskamp RMJ, Quaedflieg PJLM. (2011) Enzymatic synthesis of activated esters and their subsequent use in enzyme-based peptide synthesis. J Mol Catal B Enzym 71:79–84CrossRefGoogle Scholar
  47. 47.
    Badgujar KC, Bhanage BM (2014) Application of lipase immobilized on the biocompatible ternary blendpolymer matrix for synthesis of citronellyl acetate in non-aqueousmedia: kinetic modelling study. Enzyme Microb Technol 57:16–25CrossRefPubMedGoogle Scholar
  48. 48.
    Shi YG, Wu Y, Lu XY, Ren YP, Wang Q, Zh CM, Yu D, Wang H (2017) Lipase-catalyzed esterification of ferulic acid with lauryl alcohol in ionic liquids and antibacterial properties in vitro against three food-related bacteria. Food Chem 220:249–256CrossRefPubMedGoogle Scholar
  49. 49.
    Fallavena LP, Antunes FHF, Alves JS, Paludo N, Ayub MAZ, Fernandez-Lafuente R, Rodrigues RC (2014) Ultrasound technology and molecular sieves improve the thermodynamically controlled esterification of butyric acid mediated by immobilized lipase from Rhizomucor miehei. RSC Adv 4:8675–8681CrossRefGoogle Scholar
  50. 50.
    Cui C, Zhen Y, Qu J, Chen B, Tan T (2016) Synthesis of biosafe isosorbide dicaprylate ester plasticizer by lipase in a solvent-free system and its sub-chronic toxicity in mice. RSC Adv 6:11959–11966CrossRefGoogle Scholar
  51. 51.
    Ferrer M, Soliveri J, Plou FJ, López-Cortés N, Reyes-Duarte D, Christensen M, Copa-Patiño JL, Ballesteros A (2005) Synthesis of sugar esters in solvent mixtures by lipases from Thermomyces lanuginosus and Candida antarctica B, and their antimicrobial properties. Enzyme Microb Technol 36:391–398CrossRefGoogle Scholar
  52. 52.
    Miranda JS, Silva NCA, Bassi JJ, Corradini MCC, Lage FAP, Hirata DB, Mendes AA (2014) Immobilization of Thermomyces lanuginosus lipase on mesoporous poly-hydroxybutyrate particles and application in alkyl esters synthesis: isotherm, thermodynamic and mass transfer studies. Chem Eng J 251:392–403CrossRefGoogle Scholar
  53. 53.
    Fernandez-Lafuente R, Rosell CM, Rodriguez V, Santana C, Soler G, Bastida A, Guisan JM (1993) Preparation of activated supports containing low pK amino groups. A new tool for protein immobilization via the carboxyl coupling method. Enzyme Microb Technol 15:546–550CrossRefPubMedGoogle Scholar
  54. 54.
    Mendes AA, Castro HF, Giordano RLC (2014) Covalent attachment of lipases on glyoxyl-agarose beads: application in fruit flavor and biodiesel synthesis. Int J Biol Macromol 70:78–85CrossRefPubMedGoogle Scholar
  55. 55.
    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 72:248–254CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Bassi JJ, Todero LM, Lage FAP, Khedy GI, Ducas JD, Custódio AP, Pinto MA, Mendes AA (2016) Interfacial activation of lipases on hydrophobic support and application in the synthesis of a lubricant ester. Int J Biol Macromol 92:900–909CrossRefPubMedGoogle Scholar
  57. 57.
    Annual Book of ASTM Standards (2005) Petroleum products lubricants and fossil fuels. American Society for Testing and Materials, PhiladelphiaGoogle Scholar
  58. 58.
    Guncheva MH, Zhiryakova D (2008) High-yield synthesis of wax esters catalysed by modified Candida rugosa lipase. Biotechnol Lett 30:509–512CrossRefPubMedGoogle Scholar
  59. 59.
    Chen HC, Kuo CH, Chen HH, Liu YC, Shieh CJ (2011) Optimization of enzymatic synthesis of cetyl 2-ethylhexanoate by Novozym® 435. J Am Oil Chem Soc 88:1917–1923CrossRefGoogle Scholar
  60. 60.
    Razak NNA, Annuar MSM (2015) Enzymatic synthesis of flavonoid ester: elucidation of its kinetic mechanism and equilibrium thermodynamic behavior. Ind Eng Chem Res 54:5604–5612CrossRefGoogle Scholar
  61. 61.
    Sharma S, Dograb P, Chauhanb GS, Kanwar SS (2014) Synthesis of alkyl coumarate esters by celite-bound lipase of Bacillus licheniformis SCD11501. J Mol Catal B Enzym 101:80–86CrossRefGoogle Scholar
  62. 62.
    Pan Z, Jin S, Zhang X, Zheng S, Han S, Pan L, Ying L (2016) Biocatalytic behavior of a new Aspergillus niger whole-cell biocatalyst with high operational stability during the synthesis of green biosolvent isopropyl esters. J Mol Catal B Enzym 131:10–17CrossRefGoogle Scholar
  63. 63.
    Cui C, Guan N, Xing C, Chen B, Tan T (2016) Immobilization of Yarrowia lipolytica lipase Ylip2 for the biocatalytic synthesis of phytosterol ester in a water activity controlled reactor. Colloid Surface B 146:490–497CrossRefGoogle Scholar
  64. 64.
    Manan FMA, Attan N, Zakaria Z, Keyon ASA, Wahab RA (2018) Enzymatic esterification of eugenol and benzoic acid by a novel chitosan-chitin nanowhiskers supported Rhizomucor miehei lipase: process optimization and kinetic assessments. Enzyme Microb Technol 108:42–52CrossRefPubMedGoogle Scholar
  65. 65.
    Castro HF, Oliveira PC, Pereira EB (2000) Influence of substrate partition coefficient on the performance of lipase catalyzed synthesis of critronellyl acetate by alcoholysis. Braz J Chem Eng 17:859–866CrossRefGoogle Scholar
  66. 66.
    Radzi SM, Basri M, Salleh AB, Ariff A, Mohammad R, Rahman MBA, Rahman RNZRA. (2005) High performance enzymatic synthesis of oleyl oleate using immobilised lipase from Candida antartica. Electron J Biotechnol 8:291–298CrossRefGoogle Scholar
  67. 67.
    Trivedi J, Aila M, Sharma CD, Gupta P, Kaul S (2015) Clean synthesis of biolubricant range esters using novel liquid lipase enzyme in solvent free medium. SpringerPlus 4:165CrossRefPubMedPubMedCentralGoogle Scholar
  68. 68.
    Figueiredo KCS, Salim VMM, Borges CP (2010) Ethyl oleate production by means of pervaporation-assisted esterification using heterogeneous catalysis. Braz J Chem Eng 27:609–617CrossRefGoogle Scholar
  69. 69.
    Anzenberger C, Li S, Bouzidi L, Narine SS (2016) Synthesis of waxes from vegetable oil derived self-metathesized aliphatic esters. Ind Crops Prod 89:368–375CrossRefGoogle Scholar
  70. 70.
    El Kinawy O (2004) Comparison between jojoba oil and other vegetable oils as a substitute to petroleum. Energy Sources 26:639–645CrossRefGoogle Scholar
  71. 71.
    Habashy RR, Abdel-Naim AB, Khalifa AE, Al-Azizi MM (2005) Anti-inflammatory effects of jojoba liquid wax in experimental models. Pharmacol Res 51:95–105CrossRefPubMedGoogle Scholar
  72. 72.
    Al Awad AS, Selim MYE, Zeibak AF, Moussa R (2014) Jojoba ethyl ester production and properties of ethanol blends. Fuel 124:73–75CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Letícia C. D. Lima
    • 1
  • Daniela G. C. Peres
    • 1
  • Adriano A. Mendes
    • 1
  1. 1.Institute of ChemistryFederal University of AlfenasAlfenasBrazil

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