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Novel strategy for yeast construction using δ-integration and cell fusion to efficiently produce ethanol from raw starch

  • Applied Genetics and Molecular Biotechnology
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Abstract

We developed a novel strategy for constructing yeast to improve levels of amylase gene expression and the practical potential of yeast by combining δ-integration and polyploidization through cell fusion. Streptococcus bovis α-amylase and Rhizopus oryzae glucoamylase/α-agglutinin fusion protein genes were integrated into haploid yeast strains. Diploid strains were constructed from these haploid strains by mating, and then a tetraploid strain was constructed by cell fusion. The α-amylase and glucoamylase activities of the tetraploid strain were increased up to 1.5- and tenfold, respectively, compared with the parental strain. The diploid and tetraploid strains proliferated faster, yielded more cells, and fermented glucose more effectively than the haploid strain. Ethanol productivity from raw starch was improved with increased ploidy; the tetraploid strain consumed 150 g/l of raw starch and produced 70 g/l of ethanol after 72 h of fermentation. Our strategy for constructing yeasts resulted in the simultaneous overexpression of genes integrated into the genome and improvements in the practical potential of yeasts.

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Acknowledgments

This work was supported by a Grant-in-aid from the Ministry of Environment, Japan, and was supported in part by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan.

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Authors and Affiliations

  1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, Hyogo, 657-8501, Japan

    Ryosuke Yamada, Chiaki Ogino & Akihiko Kondo

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

    Tsutomu Tanaka & Hideki Fukuda

Authors
  1. Ryosuke Yamada
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  2. Tsutomu Tanaka
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  3. Chiaki Ogino
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  4. Hideki Fukuda
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  5. Akihiko Kondo
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Correspondence to Akihiko Kondo.

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Yamada, R., Tanaka, T., Ogino, C. et al. Novel strategy for yeast construction using δ-integration and cell fusion to efficiently produce ethanol from raw starch. Appl Microbiol Biotechnol 85, 1491–1498 (2010). https://doi.org/10.1007/s00253-009-2198-y

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  • DOI: https://doi.org/10.1007/s00253-009-2198-y

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