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Metabolic engineering of erythritol production from glycerol by Yarrowia lipolytica

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Abstract

Erythritol as a four-carbon polyol has been widely used in food, pharmaceutical and daily chemical industries with characteristics of low caloric value and high chemical stability. Here, a system metabolic engineering strategy was used to increase the yield of erythritol from glycerol in Yarrowia lipolytica by enhancing the substrate transformation and restricting the by-product synthesis. Specifically, we determined that over-expression of a newly identified erythrose reductase YPR1 was able to improve the erythritol production as same as the well-known erythrose reductase ER27. Instead of its up-regulation, knockout of erythrose reductase ER10 was effective to improve erythritol synthesis. Moreover, both over-expression of YPR1 and deletion of ER10 significantly accelerated the glycerol utilization in response to high osmotic stress. To further decrease the by-product accumulation, a restriction and recycling strategy was implemented by knockout of mannitol dehydrogenase MDH2 and enhancement of arabitol dehydrogenase ADH1 and fructokinase HXK1. The engineered strain YL13 produced a titer of 25 g/L erythritol and less than 0.5 g/L mannitol and arabitol. By over-expression of transketolase TKL1, the final strain YL14 produced 28.5 g/L erythritol and none of mannitol and arabitol. This study provides a new idea for reducing the production of by-products and improving the glycerol conversion to erythritol.

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Acknowledgements

This study was supported by the National Natural Science Fund for Young Scholars (32001035), the National Key R&D Program of China (2017YFE0115600, 2019YFA0904302) and the 111 Project (B18022).

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China

    Ya-Ting Wang, Ling-Xuan Zhao, Liu-Jing Wei, Jun Chen, Zhijie Liu, Feng Liu & Qiang Hua

  2. Key Laboratory of Fermentation Engineering (Ministry of Education), Cooperative Innovation Center of Industrial Fermentation (Ministry of Education and Hubei Province), Hubei Key Laboratory of Industrial Microbiology, Hubei University of Technology, Wuhan, 430068, China

    Ya-Ting Wang, Ling-Xuan Zhao, Liu-Jing Wei, Jun Chen, Zhijie Liu, Feng Liu & Qiang Hua

  3. Shanghai Collaborative Innovation Center for Biomanufacturing Technology, Shanghai, 200237, China

    Ya-Ting Wang, Ling-Xuan Zhao, Liu-Jing Wei, Jun Chen, Zhijie Liu, Feng Liu & Qiang Hua

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  1. Ya-Ting Wang
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  2. Ling-Xuan Zhao
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Contributions

Ya-Ting Wang and Ling-Xuan Zhao designed and carried out the experiments to construct engineered strains. Liu-Jing Wei, Jun Chen and Zhijie Liu designed and carried out the fermentation. Feng Liu and Qiang Hua drafted the work and revised. Ya-Ting Wang, Ling-Xuan Zhao, Liu-Jing Wei, Jun Chen, Zhijie Liu and Feng Liu, Qiang Hua contributed to final approval of the version to be published.

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Correspondence to Qiang Hua.

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Wang, YT., Zhao, LX., Wei, LJ. et al. Metabolic engineering of erythritol production from glycerol by Yarrowia lipolytica. Biotechnol Bioproc E 29, 119–127 (2024). https://doi.org/10.1007/s12257-024-00005-9

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