Development of conditional cell lysis mutants of Saccharomyces cerevisiae as production hosts by modulating OCH1 and CHS3 expression

  • Van-Trinh Luu
  • Hye Yun Moon
  • Su Jin Yoo
  • Jin Ho Choo
  • Eun Jung Thak
  • Hyun Ah KangEmail author
Applied genetics and molecular biotechnology


The traditional yeast Saccharomyces cerevisiae has been widely used as a host for the production of recombinant proteins and metabolites with industrial potential. However, its thick and rigid cell wall presents problems for the effective recovery of products. In this study, we modulated the expression of ScOCH1, encoding the α-1,6-mannosyltransferase responsible for outer chain biosynthesis of N-glycans, and ScCHS3, encoding the chitin synthase III required for synthesis of the majority of cell wall chitin, by exploiting the repressible ScMET3 promoter. The conditional single mutants PMET3-OCH1 and PMET3-CHS3 and the double mutant PMET3-OCH1/PMET3-CHS3 showed comparable growth to the wild-type strain under normal conditions but exhibited increased sensitivity to temperature and cell wall-disturbing agents in the presence of methionine. Such conditional growth defects were fully recovered by supplementation with 1 M sorbitol. The osmotic lysis of the conditional mutants cultivated with methionine was sufficient to release the intracellularly expressed recombinant protein, nodavirus capsid protein, with up to 60% efficiency, compared to lysis by glass bead breakage. These mutant strains also showed approximately three-fold-enhanced secretion of a recombinant extracellular glycoprotein, Saccharomycopsis fibuligera β-glucosidase, with markedly reduced hypermannosylation, particularly in the PMET3-OCH1 mutants. Furthermore, a substantial increase of extracellular glutathione production, up to four-fold, was achieved with the conditional mutant yeast cells. Together, our data support that the conditional cell wall lysis mutants constructed based on the modulation of ScOCH1 and ScCHS3 expression would likely be useful hosts for the improved recovery of proteins and metabolites with industrial application.


Saccharomyces cerevisiae Conditional mutant α-1,6-Mannosyltransferase Chitin synthase III MET3 promoter 



This work was supported by grants from the National Research Foundation of Korea (NRF), NRF-2013M3A6A8073554 (Global Frontier Program for the Intelligent Synthetic Biology), NRF-2017M3C1B5019295 (STEAM Research Project), and NRF-2018R1025077 (Advanced Research Center Program). Van-Trinh Luu is a recipient of CASSY fellowship from Chung-Ang University.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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

Supplementary material

253_2019_9614_MOESM1_ESM.pdf (971 kb)
ESM 1 The online version of this article (10.1007/....) contains supplementary material, which is available to all users. (PDF 970 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Van-Trinh Luu
    • 1
  • Hye Yun Moon
    • 1
  • Su Jin Yoo
    • 1
  • Jin Ho Choo
    • 1
  • Eun Jung Thak
    • 1
  • Hyun Ah Kang
    • 1
    Email author
  1. 1.Department of Life Science, College of Natural ScienceChung-Ang UniversitySeoulSouth Korea

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