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Characterizing methanol metabolism-related promoters for metabolic engineering of Ogataea polymorpha

  • Applied Genetics and Molecular Biotechnology
  • Published:
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Abstract

Promoters play an important role in regulating gene expression, and construction of microbial cell factories requires multiple promoters for balancing the metabolic pathways. However, there are only a limited number of characterized promoters for gene expression in the methylotrophic yeast Ogataea polymorpha, which hampers the extensive harnessing of this important yeast toward a cell factory. Here we characterized the promoters of methanol utilization pathway, precursor supply pathway, and reactive oxygen species (ROS) defense system, by using a green fluorescence protein variant (GFPUV) as a quantification signal. Finally, the characterized promoters were used for tuning a fatty alcohol biosynthetic pathway in O. polymorpha and realized fatty alcohol production from methanol. This promoter box should be helpful for gene expression and pathway optimization in the methylotrophic yeast O. polymorpha.

Key Points

• 22 promoters related to methanol metabolism were characterized in O. polymorpha.

• Promoter truncation resulted shorter and compact promoters.

• Promoters with various strengths were used for regulating a fatty alcohol biosynthesis from methanol.

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Data availability

Data that supports the finding of this study are available from the corresponding author on reasonable request.

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Acknowledgements

We appreciate the helpful discussions with Wei Yu, Peng Cai, Yunxia Li, Reibing Chen, and Chunxiao Yan.

Funding

This study was funded by National Natural Science Foundation of China (21922812, 21808216 and M-0246), LiaoNing Revitalization Talents Program (XLYC1807191), DMTO research grant (grant no. DICP DMTO201701) and DICP innovation grant (grant no. DICP I201920) from Dalian Institute of Chemicals Physics, CAS.

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Authors and Affiliations

  1. Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, No. 457 Zhongshan Road, Dalian, 116023, People’s Republic of China

    Xiaoxin Zhai, Lulu Ji, Jiaoqi Gao & Yongjin J. Zhou

  2. CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, People’s Republic of China

    Xiaoxin Zhai, Jiaoqi Gao & Yongjin J. Zhou

  3. Dalian Key Laboratory of Energy Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, People’s Republic of China

    Xiaoxin Zhai, Jiaoqi Gao & Yongjin J. Zhou

  4. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Lulu Ji

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  1. Xiaoxin Zhai
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  2. Lulu Ji
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  3. Jiaoqi Gao
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Contributions

XXZ, LLJ, and JQG designed the research; XXZ and LLJ performed experiments and data analysis; YJZ conceived and supervised the project; XXZ and YJZ wrote the paper; all authors proofed the manuscript.

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Correspondence to Yongjin J. Zhou.

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Zhai, X., Ji, L., Gao, J. et al. Characterizing methanol metabolism-related promoters for metabolic engineering of Ogataea polymorpha. Appl Microbiol Biotechnol 105, 8761–8769 (2021). https://doi.org/10.1007/s00253-021-11665-5

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  • DOI: https://doi.org/10.1007/s00253-021-11665-5

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