Bioprocess and Biosystems Engineering

, Volume 34, Issue 9, pp 1133–1142 | Cite as

Kinetics and mechanism of the cutinase-catalyzed transesterification of oils in AOT reversed micellar system

  • Sara M. Badenes
  • Francisco Lemos
  • Joaquim M. S. Cabral
Original Paper
First Online:
Received:
Accepted:

Abstract

The kinetics of the enzymatic transesterification between a mixture of triglycerides (oils) and methanol for biodiesel production in a bis(2-ethylhexyl) sodium sulfosuccinate (AOT)/isooctane reversed micellar system, using recombinant cutinase from Fusarium solani pisi as a catalyst, was investigated. In order to describe the results that were obtained, a mechanistic scheme was proposed, based on the literature and on the experimental data. This scheme includes the following reaction steps: the formation of the active enzyme–substrate complex, the addition of an alcohol molecule to the complex followed by the separation of a molecule of the fatty acid alkyl ester and a glycerol moiety, and release of the active enzyme. Enzyme inhibition and deactivation effects due to methanol and glycerol were incorporated in the model. This kinetic model was fitted to the concentration profiles of the fatty acid methyl esters (the components of biodiesel), tri-, di- and monoglycerides, obtained for a 24 h transesterification reaction performed in a stirred batch reactor under different reaction conditions of enzyme and initial substrates concentration.

Keywords

Kinetic model Cutinase Biodiesel Transesterification Reversed micelle 

List of symbols

TG

Triglycerides

DG

Diglycerides

MG

Monoglycerides

A

Methanol (alcohol)

AE

Fatty acid methyl ester (alkyl ester)

G

Glycerol

ETG, EDG, EMG

Active enzyme-substrate complexes (with tri-, di- and monoglycerides)

EA

Dead-end enzyme-alcohol complex

EG

Dead-end enzyme-glycerol complex

ET

Total enzyme

E

Free enzyme

E0

Active enzyme, state 0

E1

Active enzyme, state 1

E2

Enzyme denatured, state 2

\( k_{1} ,\;k_{ - 1} ,\;k_{2} ,\;k_{ - 2} ,\;k_{3} ,\;k_{ - 3} \)

Reaction rate constants

\( K_{4} ,\;K_{5} ,\;K_{6} \)

Coordination constants

K7, K8

Inhibition coordination constants

\( k_{\text{d1}}^{\text{aot}} ,k_{\text{d2}}^{\text{aot}} ,k_{\text{d1}}^{\text{met}} ,k_{\text{d2}}^{\text{met}} \)

Deactivation constants

This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

S. M. Badenes acknowledges a PhD grant (SFRH/BD/28895/2006) from Fundação para a Ciência e Tecnologia.

References

  1. 1.
    Antczak MS, Kubiak A, Antczak T, Bielecki S (2009) Renew Energ 34:1185–1194CrossRefGoogle Scholar
  2. 2.
    Manzanera M, Molina-Muñoz ML, González-López J (2008) Recent Patents Biotechnol 2:25–34CrossRefGoogle Scholar
  3. 3.
    Fukuda H, Kondo A, Noda H (2001) J Biosci Bioeng 92:405–416CrossRefGoogle Scholar
  4. 4.
    Ranganathan SV, Narasimhan SL, Muthukumar K (2008) Bioresour Technol 99:3975–3981CrossRefGoogle Scholar
  5. 5.
    Carvalho CML, Aires-Barros MR, Cabral JMS (1999) Biotechnol Bioeng 66:17–34CrossRefGoogle Scholar
  6. 6.
    Lauwereys M, de Geus P, de Meutter J, Stanssens P and Matthyssens G (1990) In: Alberghirna L SR, Verger R (ed) Lipases structure, mechanism and genetic engineering. GBF Monographs, BraunschwergGoogle Scholar
  7. 7.
    Dutta K, Sen S, Veeranki VD (2009) Process Biochem 44:127–134CrossRefGoogle Scholar
  8. 8.
    Badenes SM, Lemos F, Cabral JMS (2010) Biotechnol Lett 32:399–403CrossRefGoogle Scholar
  9. 9.
    Melo EP, Aires-Barros MR, Cabral JMS (1995) Appl Biochem Biotechnol 50:45–56CrossRefGoogle Scholar
  10. 10.
    Prazeres DMF, Lemos F, Garcia FAP, Cabral JMS (1993) Biotechnol Bioeng 42:759–764CrossRefGoogle Scholar
  11. 11.
    Sebastião MJ, Cabral JMS, Aires-Barros MR (1993) Biotechnol Bioeng 42:326–332CrossRefGoogle Scholar
  12. 12.
    Schlatmann J, Aires-Barros MR, Cabral JMS (1991) Biocatalysis 5:137–144CrossRefGoogle Scholar
  13. 13.
    Cunnah PJ, AiresBarros MR, Cabral JMS (1996) Biocatal Biotransformation 14:125–146CrossRefGoogle Scholar
  14. 14.
    Carvalho CML, Serralheiro MLM, Cabral JMS, Aires-Barros MR (1997) Enzyme Microb Technol 21:117–123CrossRefGoogle Scholar
  15. 15.
    de Barros DPC, Lemos F, Fonseca LP, Cabral JMS (2010) J Mol Catal B Enzym 66:285–293CrossRefGoogle Scholar
  16. 16.
    Carvalho CML, Cabral JMS, Aires-Barros MR (1999) Enzyme Microb Technol 24:569–576CrossRefGoogle Scholar
  17. 17.
    Bru R, Sánchez-Ferrer A, García-Carmona F (1995) Biochem J 310:721–739Google Scholar
  18. 18.
    Carvalho CML, Cabral JMS (2000) Biochimie 82:1063–1085CrossRefGoogle Scholar
  19. 19.
    Mukherjee S, Miller CA, Fort JT (1983) J Coll Inter Sci 91:223–243CrossRefGoogle Scholar
  20. 20.
    Badenes SM, Lemos F, Cabral JMS (2011) J Chem Technol Biotechnol 86:34–41CrossRefGoogle Scholar
  21. 21.
    Henley JP, Sadana A (1985) Enzyme Microb Technol 7:50–60CrossRefGoogle Scholar
  22. 22.
    Melo EP, Carvalho CML, Aires-Barros MR, Costa SMB, Cabral JMS (1998) Biotechnol Bioeng 58:380–386CrossRefGoogle Scholar
  23. 23.
    Pocalyko DJ, Tallman M (1998) Enzyme Microb Technol 22:647–651CrossRefGoogle Scholar
  24. 24.
    Badenes SM, Lemos F, Cabral JMS (2010) J Chem Technol Biotechnol 85:993–998CrossRefGoogle Scholar
  25. 25.
    Melo EP, Fojan P, Cabral JMS, Petersen SB (2000) Chem Phys Lipids 106:181–189CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Sara M. Badenes
    • 1
  • Francisco Lemos
    • 1
  • Joaquim M. S. Cabral
    • 1
  1. 1.IBB, Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior TécnicoTechnical University of LisbonLisbonPortugal

Personalised recommendations