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Increased production of fatty acids and triglycerides in Aspergillus oryzae by enhancing expressions of fatty acid synthesis-related genes

  • Applied genetics and molecular biotechnology
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Abstract

Microbial production of fats and oils is being developed as a means of converting biomass to biofuels. Here we investigate enhancing expression of enzymes involved in the production of fatty acids and triglycerides as a means to increase production of these compounds in Aspergillus oryzae. Examination of the A. oryzae genome demonstrates that it contains two fatty acid synthases and several other genes that are predicted to be part of this biosynthetic pathway. We enhanced the expression of fatty acid synthesis-related genes by replacing their promoters with the promoter from the constitutively highly expressed gene tef1. We demonstrate that by simply increasing the expression of the fatty acid synthase genes we successfully increased the production of fatty acids and triglycerides by more than two-fold. Enhancement of expression of the fatty acid pathway genes ATP-citrate lyase and palmitoyl-ACP thioesterase increased productivity to a lesser extent. Increasing expression of acetyl-CoA carboxylase caused no detectable change in fatty acid levels. Increases in message level for each gene were monitored using quantitative real-time reverse transcription polymerase chain reaction. Our data demonstrate that a simple increase in the abundance of fatty acid synthase genes can increase the detectable amount of fatty acids.

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Acknowledgments

We appreciate the daily research-related paperwork by Ms. Debra Lee and Ms. Ana Lundeby at PNNL. We are also grateful to Dr. Osamu Yamada (National Research Institute of Brewing, Japan) for giving us information on the DNA sequences of mutations at niaD and sC loci, respectively, of A. oryzae RIB40 niaD− sC− strain that is called NS4 generally. And we thank Dr. Katsuya Gomi (Tohoku University) for providing A. oryzae RIB40 ΔligD::ptrA niaD− sC− strain, a derivative of NS4, to construct both NS4DLP and NS4DLDP. This work was performed by the grant-in-aid of the Ministry of Economy, Trade, and Industry, Japan. Also this work was supported by the grant-in-aid of the Department of Energy, USA.

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  1. Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo, Hokkaido, 062-8517, Japan

    Koichi Tamano, Myco Umemura & Masayuki Machida

  2. Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, Higashi 1-1-1, Tsukuba, Ibaraki, 305-8566, Japan

    Koichi Tamano, Myco Umemura, Hideaki Koike & Masayuki Machida

  3. Pacific Northwest National Laboratory, 902 Battelle Boulevard, Richland, WA, 99352, USA

    Kenneth S. Bruno, Sue A. Karagiosis, David E. Culley, Shuang Deng, James R. Collett & Scott E. Baker

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  1. Koichi Tamano
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  2. Kenneth S. Bruno
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  3. Sue A. Karagiosis
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  10. Masayuki Machida
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Correspondence to Masayuki Machida.

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Tamano, K., Bruno, K.S., Karagiosis, S.A. et al. Increased production of fatty acids and triglycerides in Aspergillus oryzae by enhancing expressions of fatty acid synthesis-related genes. Appl Microbiol Biotechnol 97, 269–281 (2013). https://doi.org/10.1007/s00253-012-4193-y

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