Biotechnology and Bioprocess Engineering 2024; 29(1): 119-127  
Metabolic engineering of erythritol production from glycerol by Yarrowia lipolytica
Ya‑Ting Wang1,2,3 · Ling‑Xuan Zhao1,2,3 · Liu‑Jing Wei1,2,3 · Jun Chen1,2,3 · Zhijie Liu1,2,3 · Feng Liu1,2,3 · Qiang Hua1,2,3
1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
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
3 Shanghai Collaborative Innovation Center for Biomanufacturing Technology, Shanghai 200237, China
Correspondence to: Qiang Hua
qhua@ecust.edu.cn
Received: July 4, 2023; Revised: September 18, 2023; Accepted: October 22, 2023; Published online: February 16, 2024.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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.
Keywords: Erythritol · By-products · Yarrowia lipolytica · Metabolic engineering

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