Metabolic engineering of erythritol production from glycerol by Yarrowia lipolytica

IF 2.5 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology and Bioprocess Engineering Pub Date : 2024-02-16 DOI:10.1007/s12257-024-00005-9

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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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脂肪分解亚罗酵母利用甘油生产赤藓糖醇的代谢工程

摘要赤藓糖醇作为一种四碳多元醇，具有热值低、化学稳定性高的特点，被广泛应用于食品、医药和日化行业。本文采用系统代谢工程策略，通过加强底物转化和限制副产物合成，提高了脂肪分解亚罗威氏菌（Yarrowia lipolytica）从甘油中提取赤藓糖醇的产量。具体来说，我们发现过度表达新发现的赤藓酮糖还原酶 YPR1 与众所周知的赤藓酮糖还原酶 ER27 一样，都能提高赤藓糖醇的产量。敲除赤藓酮糖还原酶 ER10 并不能上调赤藓酮糖还原酶的活性，反而能有效改善赤藓糖醇的合成。此外，在高渗透压胁迫下，过表达 YPR1 和缺失 ER10 都能显著加速甘油的利用。为了进一步减少副产物的积累，通过敲除甘露醇脱氢酶 MDH2 和增强阿拉伯糖醇脱氢酶 ADH1 和果糖激酶 HXK1，实施了限制和循环策略。改造菌株 YL13 产生的赤藓糖醇滴度为 25 克/升，甘露醇和阿拉伯糖醇滴度低于 0.5 克/升。通过过度表达转酮醇酶 TKL1，最终的菌株 YL14 产生了 28.5 克/升赤藓糖醇，没有产生甘露醇和阿拉伯糖醇。这项研究为减少副产品的产生和提高甘油转化为赤藓糖醇提供了一个新思路。

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来源期刊

Biotechnology and Bioprocess Engineering 工程技术-生物工程与应用微生物

CiteScore

5.00

自引率

12.50%

发文量

审稿时长

3 months

期刊介绍： Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.