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Abolishment of N-glycan mannosylphosphorylation in glyco-engineered Saccharomyces cerevisiae by double disruption of MNN4 and MNN14 genes

  • Applied genetics and molecular biotechnology
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Abstract

Mannosylphosphorylated glycans are found only in fungi, including yeast, and the elimination of mannosylphosphates from glycans is a prerequisite for yeast glyco-engineering to produce human-compatible glycoproteins. In Saccharomyces cerevisiae, MNN4 and MNN6 genes are known to play roles in mannosylphosphorylation, but disruption of these genes does not completely remove the mannosylphosphates in N-glycans. This study was performed to find unknown key gene(s) involved in N-glycan mannosylphosphorylation in S. cerevisiae. For this purpose, each of one MNN4 and five MNN6 homologous genes were deleted from the och1Δmnn1Δmnn4Δmnn6Δ strain, which lacks yeast-specific hyper-mannosylation and the immunogenic α(1,3)-mannose structure. N-glycan profile analysis of cell wall mannoproteins and a secretory recombinant protein produced in mutants showed that the MNN14 gene, an MNN4 paralog with unknown function, is essential for N-glycan mannosylphosphorylation. Double disruption of MNN4 and MNN14 genes was enough to eliminate N-glycan mannosylphosphorylation. Our results suggest that the S. cerevisiae och1Δmnn1Δmnn4Δmnn14Δ strain, in which all yeast-specific N-glycan structures including mannosylphosphorylation are abolished, may have promise as a useful platform for glyco-engineering to produce therapeutic glycoproteins with human-compatible N-glycans.

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Authors and Affiliations

  1. Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, South Korea

    Yeong Hun Kim, Ji-Yeon Kang, Jin Young Gil, Sang-Yoon Kim, Keun Koo Shin, Ohsuk Kwon & Doo-Byoung Oh

  2. Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon, South Korea

    Yeong Hun Kim & Jeong-Yoon Kim

  3. Department of Life Science, Chung-Ang University, Seoul, South Korea

    Hyun Ah Kang

  4. Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon, South Korea

    Ohsuk Kwon & Doo-Byoung Oh

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  1. Yeong Hun Kim
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  2. Ji-Yeon Kang
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  3. Jin Young Gil
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  4. Sang-Yoon Kim
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  9. Doo-Byoung Oh
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Correspondence to Ohsuk Kwon or Doo-Byoung Oh.

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Funding

This study was funded by the National Research Foundation of Korea [NRF-2013M3A9B6075888], the Next-Generation BioGreen 21 Program [PJ011078] of the Rural Development Administration, and Korea Research Institute of Bioscience and Biotechnology (KRIBB) in Republic of Korea.

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The authors declare that they have no conflict of interest.

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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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Yeong Hun Kim and Ji-Yeon Kang contributed equally to this work.

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Kim, Y.H., Kang, JY., Gil, J.Y. et al. Abolishment of N-glycan mannosylphosphorylation in glyco-engineered Saccharomyces cerevisiae by double disruption of MNN4 and MNN14 genes. Appl Microbiol Biotechnol 101, 2979–2989 (2017). https://doi.org/10.1007/s00253-017-8101-3

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  • DOI: https://doi.org/10.1007/s00253-017-8101-3

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