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Efficient and simple electro-transformation of intact cells for the basidiomycetous fungus Pseudozyma hubeiensis

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Abstract

Objective

An electroporation procedure for the species was investigated to develop an efficient transformation method for the basidiomycetous fungus Pseudozyma hubeiensis SY62, a strong biosurfactant-producing host.

Results

A plasmid, pUXV1emgfp including green fluorescence protein as a reporter gene, was constructed to determine the transformation and expression of foreign genes. Optimal electroporation conditions achieved 44.8 transformants μg−1 plasmid competency (intact cells) without protoplast treatment. Lithium acetate treatments increased the efficiency to approx. Twice that of control experiments. Almost all transformants demonstrated green fluorescence expressed in the transformant cells.

Conclusion

The optimal method, successfully applied to several related species, yields sufficient transformant colonies to engineer the host strain.

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Acknowledgments

This work was financially supported by JSPS KAKENHI Grant Numbers 24681013. We thank to JAMSTEC for kindly providing Pseudozyma hubeiensis SY62.

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Authors and Affiliations

  1. Development of Biotechnology and Environmental Chemistry, Kitami Institute of Technology, 165 Koen-cho, Kitami, Hokkaido, 090-8507, Japan

    Masaaki Konishi, Yuta Yoshida, Mizuki Ikarashi & Jun-ichi Horiuchi

  2. Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15, Natsushima-cho, Yokosuka, 237-0061, Japan

    Masaaki Konishi

Authors
  1. Masaaki Konishi
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  2. Yuta Yoshida
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  3. Mizuki Ikarashi
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  4. Jun-ichi Horiuchi
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Correspondence to Masaaki Konishi.

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Konishi, M., Yoshida, Y., Ikarashi, M. et al. Efficient and simple electro-transformation of intact cells for the basidiomycetous fungus Pseudozyma hubeiensis . Biotechnol Lett 37, 1679–1685 (2015). https://doi.org/10.1007/s10529-015-1837-x

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  • DOI: https://doi.org/10.1007/s10529-015-1837-x

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