Skip to main content
SpringerLink
Log in

Preparation of immobilized whole cell biocatalyst and biodiesel production using a packed-bed bioreactor

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

Rhizopus oryzae NBRC 4697 was selected from among promising candidates as a biocatalyst for biodiesel production. This microorganism was immobilized on to polyurethane foam coated with activated carbon for reuse, and, for biodiesel production. Vacuum drying of the immobilized cells was found to be more efficient than natural or freeze-drying processes. Although the immobilized cells were severely inhibited by a molar ratio of methanol to soybean oil in excess of 2.0, stepwise methanol addition (3 aliquots at 24-h feeding intervals) significantly prevented methanol inhibition. A packed-bed bioreactor (PBB) containing the immobilized whole cell biocatalyst was then operated under circulating batch mode. Stepwise methanol feeding was used to mitigate methanol inhibition of the immobilized cells in the PBB. An increase in the feeding rate (circulating rate) of the reaction mixture barely affected biodiesel production, while an increase in the packing volume of the immobilized cells enhanced biodiesel production noticeably. Finally, repeated circulating batch operation of the PBB was carried out for five consecutive rounds without a noticeable decrease in the performance of the PBB for the three rounds.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Antoni D, Zverlow VV, Schwarz WH (2007) Biofuels from microbes. Appl Microbiol Biotechnol 77:23–35

    Article  CAS  Google Scholar 

  2. Jeon DJ, Yeom SH (2010) Two-step bioprocess employing whole cell and enzyme for economical biodiesel production. Korean J Chem Eng 27:1555–1559

    Article  CAS  Google Scholar 

  3. Atadashi IM, Aroua MK, Abdul Aziz AR, Sulaiman NMN (2013) The effects of catalysts in biodiesel production: a review. J Ind Eng Chem 19:14–26

    Article  CAS  Google Scholar 

  4. Fukuda H, Hama S, Tamalampudi S, Noda H (2008) Whole-cell biocatalysts for biodiesel fuel production. Trends Biotechnol 26:668–673

    Article  CAS  Google Scholar 

  5. Ban K, Hama S, Nishizuka K, Kaieda M, Matsumoto T, Kondo A, Noda H, Fukuda H (2002) Repeated use of whole-cell biocatalysts immobilized within biomass support particles for biodiesel fuel production. J Mol Catal B Enzym 17:157–165

    Article  CAS  Google Scholar 

  6. Ranganathan SV, Marasimhan L, Muthukumar K (2008) An overview of enzymatic production of biodiesel. Bioresour Technol 99:3975–3981

    Article  CAS  Google Scholar 

  7. Shen L, Yin H, Wang A, Lu X, Zhang C, Chen F, Wang Y, Chen H (2014) Liquid phase catalytic dehydration of glycerol to acrolein over Bronsted acidic ionic liquid catalysts. J Ind Eng Chem 20:759–766

    Article  CAS  Google Scholar 

  8. Li L, Du W, Liu D, Wang L, Li Z (2006) Lipase-catalyzed transesterification of rapeseed oils for biodiesel production with a novel organic solvent as the reaction medium. J Mol Catal B Enzym 43:58–62

    Article  CAS  Google Scholar 

  9. Jeon DJ, Yeom SH (2011) Comparison of methods for preventing methanol inhibition in enzymatic production of biodiesel. Korean J Chem Eng 28:1420–1426

    Article  CAS  Google Scholar 

  10. Fjerbaek L, Christensen KV, Norddahl B (2009) A review of the current state of biodiesel production using enzymatic transesterification. Biotechnol Bioeng 102:1298–1315

    Article  CAS  Google Scholar 

  11. Shimada Y, Watanabe Y, Suglihara A, Tominaga Y (2002) Enzymatic alcoholysis for biodiesel fuel production and application of the reaction to oil processing. J Mol Catal B Enzym 17:133–142

    Article  CAS  Google Scholar 

  12. Samukawa T, Kaieda M, Matsumoto T, Ban K, Kondo A, Shimada Y, Noda H, Fukuda H (2000) Pretreatment of Immobilized Candida antarctica lipase for Biodiesel fuel production from plant oil. J Bioresour Bioeng 90:180–183

    Article  CAS  Google Scholar 

  13. Hama S, Yamaji H, Fukumizu T, Numata T, Tamalampudi S, Kondo A, Noda H, Fukuda H (2007) Biodiesel-fuel production in a packed-bed reactor using lipase-producing Rhizopus oryzae cells immobilized within biomass support particles. Biochem Eng J 34:273–278

    Article  CAS  Google Scholar 

  14. Talukder MMR, Wu JC, van Nguyen TB, Fen NM, Melissa YLS (2009) Novozym 435 for production of biodiesel from unrefined palm oil: comparison of methanolysis methods. J Mol Catal B Enzym 60:106–112

    Article  CAS  Google Scholar 

  15. Chen HC, Ju HY, Wu TT, Liu YC, Lee CC, Chang C, Chung YL, Shieh CJ (2011) Continuous production of lipase-catalyzed biodiesel in a packed-bed reactor: optimization and enzyme reuse study. J Biomed Biotechnol. doi:10.1155/2011/950725

    Google Scholar 

  16. Go YW, Yeom SH (2014) Application of pseudo-two phase partitioning bioreactor (P-TPPB) to the production of biodiesel. Biopro Biosys Eng 37:269–275

    Article  CAS  Google Scholar 

  17. Jin G, Bierma TJ, Hamaker CG, Mucha R, Schola V, Stewart J, Wade C (2009) Use of a whole-cell biocatalyst to produce biodiesel in a water-containing system. J Environ Sci Health Part A 44:21–28

    Article  CAS  Google Scholar 

  18. Jin G, Bierma TJ, Hamaker CG, Rhykerd R, Loftus LA (2008) Producing biodiesel using whole-cell biocatalysts in separate hydrolysis and methanolysis reactions. J Environ Sci Health Part A 43:589–595

    Article  CAS  Google Scholar 

  19. Arai S, Nakashima K, Tanino T, Ogino C, Kondo A, Fukuda H (2010) Production of biodiesel fuel from soybean oil catalyzed by fungus whole-cell biocatalysts in ionic liquids. Enzym Microb Technol 46:51–55

    Article  CAS  Google Scholar 

  20. Hama S, Tamalampudi S, Fukumizu T, Miura K, Yamaji H, Kondo A, Fukuda H (2006) Lipase localization in Rhizopus oryzae cells immobilized within biomass support particles for use as whole-cell biocatalysts in biodiesel-fuel production. J Biosci Bioeng 101:328–333

    Article  CAS  Google Scholar 

  21. Tamalampudi S, Talukder MR, Hama S, Numata T, Kondo A, Fukuda H (2008) Enzymatic production of biodiesel from Jatropha oil: a comparative study of immobilized-whole cell and commercial lipases as a biocatalyst. Biochem Eng J 39:185–189

    Article  CAS  Google Scholar 

  22. Li W, Du W, Liu D (2007) Rhizopus oryzae IFO 4697 whole cell catalyzed methanolysis of crude and acidified rapeseed oils for biodiesel production in tert-butanol system. Process Biochem 42:1481–1485

    Article  CAS  Google Scholar 

  23. Sun T, Du W, Liu D (2011) Comparative study on stability of whole cells during biodiesel production in solvent-free system. Process Biochem 46:661–664

    Article  CAS  Google Scholar 

  24. Matsumoto T, Takahashi S, Kaieda M, Ueda M, Tanaka A, Fukuda H, Kondo A (2001) Yeast whole-cell biocatalyst constructed by intracellular overproduction of Rhizopus oryzae lipase is applicable to biodiesel fuel production. Appl Microb Biotechnol 57:515–529

    Article  CAS  Google Scholar 

  25. Du W, Li W, Sun T, Chen X, Liu D (2008) Perspectives for biotechnological production of biodiesel and impacts. Appl Microb Biotechnol 79:331–337

    Article  CAS  Google Scholar 

  26. Yoshida A, Hama S, Tamadani N, Fukuda H, Kondo A (2012) Improved performance of a packed-bed reactor for biodiesel production through whole-cell biocatalysis employing a high-lipase-expression system. Biochem Eng J 63:76–80

    Article  CAS  Google Scholar 

  27. Yoshida A, Hama S, Tamadani N, Noda H, Fukuda H, Kondo A (2012) Continuous production of biodiesel using whole-cell biocatalyst: sequential conversion of an aqueous oil emulsion into anhydrous product. Biochem Eng J 68:7–11

    Article  CAS  Google Scholar 

  28. Hama S, Tamalampudi S, Suzuki Y, Yoshida A, Fukuda H, Kondo A (2008) Preparation and comparative characterization of immobilized Aspergillus oryzae expressing Fusarium heterosporum lipase for enzymatic biodiesel production. Appl Microb Biotechnol 81:637–645

    Article  CAS  Google Scholar 

  29. Xu Y, Du W, Zeng J, Liu D (2004) Conversion of soybean oil to biodiesel fuel using lipozyme TL IM in a solvent-free medium. Biocatal Biotrans 22:45–48

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by Gangwon-do, Korea and the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010-0025572). We greatly appreciate the support. The authors also would like to thank Pansuwan Supaporn and Young Wook Go for their technical assistance.

Author information

Authors and Affiliations

  1. Department of Biochemical Engineering, Gangneung-Wonju National University, Gangneung, Gangwon-do, 210-702, Korea

    Jin Seon Kyeong & Sung Ho Yeom

Authors
  1. Jin Seon Kyeong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sung Ho Yeom
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sung Ho Yeom.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kyeong, J.S., Yeom, S.H. Preparation of immobilized whole cell biocatalyst and biodiesel production using a packed-bed bioreactor. Bioprocess Biosyst Eng 37, 2189–2198 (2014). https://doi.org/10.1007/s00449-014-1196-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-014-1196-3

Keywords

Search

Navigation