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Enhanced production of a diastereomer type of mannosylerythritol lipid-B by the basidiomycetous yeast Pseudozyma tsukubaensis expressing lipase genes from Pseudozyma antarctica

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Abstract

Basidiomycetous yeasts in the genus Pseudozyma are known to produce extracellular glycolipids called mannosylerythritol lipids (MELs). Pseudozyma tsukubaensis produces a large amount of MEL-B using olive oil as the sole carbon source (> 70 g/L production). The MEL-B produced by P. tsukubaensis is a diastereomer type of MEL-B, which consists of 4-O-β-d-mannopyranosyl-(2R,3S)-erythritol as a sugar moiety, in contrast to the conventional type of MELs produced by P. antarctica, which contain 4-O-β-d mannopyranosyl-(2S,3R)-erythritol. In this study, we attempted to increase the production of the diastereomer type of MEL-B in P. tsukubaensis 1E5 by introducing the genes encoding two lipases, PaLIPAp (PaLIPA) and PaLIPBp (PaLIPB) from P. antarctica T-34. Strain 1E5 expressing PaLIPA exhibited higher lipase activity than the strain possessing an empty vector, which was used as a negative control. Strains of 1E5 expressing PaLIPA or PaLIPB showed 1.9- and 1.6-fold higher MEL-B production than the negative control strain, respectively, and oil consumption was also accelerated by the introduction of these lipase genes. MEL-B production was estimated using time course analysis in the recombinant strains. Strain 1E5 expressing PaLIPA produced 37.0 ± 1.2 g/L of MEL-B within 4 days of cultivation, whereas the strain expressing an empty vector produced 22.1 ± 7.5 g/L in this time. Overexpression of PaLIPA increased MEL-B production by P. tsukubaensis strain 1E5 from olive oil as carbon source by more than 1.7-fold.

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Acknowledgements

This work has been supported by Japan Science and Technology Agency (JST) under Adaptable and Seamless Technology Transfer Program through Target-driven R&D.

Funding

This work was funded by Japan Science and Technology Agency (JST) under Adaptable and Seamless Technology Transfer Program through Target-driven R&D (AS2621413N).

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Authors and Affiliations

  1. Research Institute for Sustainable Chemistry, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 5-2, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8565, Japan

    Azusa Saika & Tomotake Morita

  2. Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6-9, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan

    Hideaki Koike

  3. Toyobo Co., Ltd. Tsuruga Institute of Biotechnology, 10-24, Toyo-cho, Tsuruga, Fukui, 914-8550, Japan

    Shuhei Yamamoto & Takahide Kishimoto

Authors
  1. Azusa Saika
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  2. Hideaki Koike
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  3. Shuhei Yamamoto
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  4. Takahide Kishimoto
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  5. Tomotake Morita
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Correspondence to Tomotake Morita.

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Conflict of interest

SY and TK are employees of Toyobo Co., Ltd., but Toyobo Co., Ltd. did not have any role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. AS, HK, TF, SY, TK, and TM are inventors of submitted patent which related to this study (JP2016-112087). The authors have declared that no other competing interests exist.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Saika, A., Koike, H., Yamamoto, S. et al. Enhanced production of a diastereomer type of mannosylerythritol lipid-B by the basidiomycetous yeast Pseudozyma tsukubaensis expressing lipase genes from Pseudozyma antarctica . Appl Microbiol Biotechnol 101, 8345–8352 (2017). https://doi.org/10.1007/s00253-017-8589-6

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  • DOI: https://doi.org/10.1007/s00253-017-8589-6

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