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Preparation and comparative characterization of immobilized Aspergillus oryzae expressing Fusarium heterosporum lipase for enzymatic biodiesel production

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Abstract

In this paper, we provide the first report of utilizing recombinant fungal whole cells in enzymatic biodiesel production. Aspergillus oryzae, transformed with a heterologous lipase-encoding gene from Fusarium heterosporum, produced fully processed and active forms of recombinant F. heterosporum lipase (FHL). Cell immobilization within porous biomass support particles enabled the convenient usage of FHL-producing A. oryzae as a whole-cell biocatalyst for lipase-catalyzed methanolysis. The addition of 5% water to the reaction mixture was effective in both preventing the lipase inactivation by methanol and facilitating the acyl migration in partial glycerides, resulting in the final methyl ester content of 94% even in the tenth batch cycle. A comparative study showed that FHL-producing A. oryzae attained a higher final methyl ester content and higher lipase stability than Rhizopus oryzae, the previously developed whole-cell biocatalyst. Although both FHL and R. oryzae lipase exhibit 1,3-regiospecificity towards triglyceride, R. oryzae accumulated a much higher amount of sn−2 isomers of partial glycerides, whereas FHL-producing A. oryzae maintained a low level of the sn−2 isomers. This is probably because FHL efficiently facilitates the acyl migration from the sn−2 to the sn−1(3) position in partial glycerides. These findings indicate that the newly developed FHL-producing A. oryzae is an effective whole-cell biocatalyst for enzymatic biodiesel production.

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Acknowledgements

This work was partly supported by the Japanese Ministry of the Environment for Technical Development of Measures to Prevent Global Warming (2007). Generous gifts of the fungal expression vector pNAN8142 and Aspergillus oryzae niaD300 from Ozeki (Hyogo, Japan) are gratefully acknowledged. We also thank Dr. Yuji Shimada (Osaka Municipal Technical Research Institute, Osaka, Japan) for providing pYGF2.

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Authors and Affiliations

  1. Bio-energy Corporation, Research and Development Laboratory, 2-9-7 Minaminanamatsu, Amagasaki, 660-0053, Japan

    Shinji Hama, Sriappareddy Tamalampudi & Ayumi Yoshida

  2. Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Sriappareddy Tamalampudi, Ayumi Yoshida & Hideki Fukuda

  3. Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Yuya Suzuki & Akihiko Kondo

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  1. Shinji Hama
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  2. Sriappareddy Tamalampudi
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  3. Yuya Suzuki
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  4. Ayumi Yoshida
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  5. Hideki Fukuda
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  6. Akihiko Kondo
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Correspondence to Akihiko Kondo.

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Hama, S., Tamalampudi, S., Suzuki, Y. et al. Preparation and comparative characterization of immobilized Aspergillus oryzae expressing Fusarium heterosporum lipase for enzymatic biodiesel production. Appl Microbiol Biotechnol 81, 637–645 (2008). https://doi.org/10.1007/s00253-008-1689-6

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  • DOI: https://doi.org/10.1007/s00253-008-1689-6

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