Engineering of Yarrowia lipolytica for producing pyruvate from glycerol

Engineering of Yarrowia lipolytica for producing pyruvate from glycerol
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解脂耶氏酵母从甘油生产丙酮酸的工程

DOI:
10.1007/s13205-022-03158-7
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发表时间:
2022-03
期刊:
影响因子:
2.8
通讯作者:
Wei Yuan
Wei Yuan
中科院分区:
工程技术4区
文献类型:
--
作者:
Songmao Wang;Yuanyuan Yang;Kechen Yu;Shiyi Xu;Mengzhu Liu;Jie Sun;Jianyong Zheng;Yinjun Zhang;Wei Yuan

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本研究旨在通过改造解脂耶氏酵母的甘油代谢途径来增加丙酮酸产量。结果:野生型解脂耶氏酵母(Po1d)经工程改造产生了6种不同的菌株,分别是ZS099(通过过表达PYK1)、ZS100(通过删除DGA2)、ZS101(通过过表达DAK1、DAK2和GCY1)、ZS102(通过过表达GUT1和GUT2)、ZS103(通过过表达GUT1和GUT2)。 GUT1)和ZSGP(通过过度表达POS5并删除GPD2)。使用高效液相色谱 (HPLC) 测定工程菌株和对照菌株的丙酮酸产量。随后对生产丙酮酸的发酵条件进行了优化,包括初始接种量、碳酸钙(CaCO3)、硫胺素和甘油的添加量。最后,为了扩大规模,使用了 20 L 发酵罐。据观察,与对照菌株(3.62 g/L)相比,ZSGP 菌株的丙酮酸产量增加了 136%(8.55 g/L)。此外,在放大工艺中,ZSGP 的丙酮酸产量在 96 小时内达到了 110.4 g/L。我们得出结论,Y 的 ZSGP 菌株。 lipolytica 可有效用于工业水平的丙酮酸生产。补充信息:在线版本包含可在 10.1007/s13205-022-03158-7 获取的补充材料。
The present study aims to increase pyruvate production by engineeringYarrowia lipolyticathrough modifying the glycerol metabolic pathway. Results: Wild-typeYarrowia lipolytica(Po1d) was engineered to produce six different strains, namely ZS099 (by over-expressing PYK1), ZS100 (by deleting DGA2), ZS101 (by over-expressing DAK1, DAK2, and GCY1), ZS102 (by over-expressing GUT1 and GUT2), ZS103 (by over-expressing GUT1) and ZSGP (by over-expressing POS5 and deleting GPD2). Production of pyruvate from engineered and control strains was determined using high-performance liquid chromatography (HPLC). Subsequently, the fermentation conditions for producing pyruvate were optimized, including the amount of initial inoculation, the addition of calcium carbonate (CaCO3), thiamine and glycerol. Finally, for scaled-up purposes, a 20-L fermentor was used. It was observed that pyruvate production increased by 136% (8.55 g/L) in ZSGP strain compared to control (3.62 g/L). Furthermore, pyruvate production by ZSGP reached up to 110.4 g/L in 96 h in the scaled-up process. We conclude that ZSGP strain ofY. lipolyticacan be effectively used for pyruvate production at the industrial level.Supplementary information:The online version contains supplementary material available at 10.1007/s13205-022-03158-7.
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