was successfully engineered to synthesize erythritol from crude glycerol, a cheap by-product of biodiesel production, but the yield remained low. Here, a biosensor-guided adaptive evolution screening platform was constructed to obtain mutant strains which could efficiently utilize crude glycerol to produce erythritol. Erythrose reductase D46A (M1) was identified as a key mutant through whole-genome sequencing of the strain G12, which exhibited higher catalytic activity (1.6-fold of the wild-type). M1 was further modified to obtain a combinatorial mutant with 4.1-fold enhancement of catalytic activity. Finally, the metabolic network was reconfigured to redirect carbon fluxes toward erythritol synthesis. The erythritol titer of the engineered strain G31 reached 220.5 g/L with a productivity of 1.8 g/L/h in a 5-L bioreactor. The study provides valuable guidance for biosensor-based ultra-high-throughput screening strategies in , as well as presenting a new paradigm for the sustainable valorization of crude glycerol.

中文翻译：

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解脂耶氏酵母高产赤藓糖醇的多维组合筛选


已成功地从粗甘油（生物柴油生产的廉价副产品）中合成赤藓糖醇，但产量仍然很低。本研究构建了生物传感器引导的适应性进化筛选平台，以获得能够高效利用粗甘油生产赤藓糖醇的突变菌株。通过菌株G12的全基因组测序，鉴定出赤藓糖还原酶D46A（M1）为关键突变体，其催化活性较高（是野生型的1.6倍）。对M1进行进一步修饰，得到催化活性增强4.1倍的组合突变体。最后，代谢网络被重新配置，将碳通量重新导向赤藓糖醇合成。工程菌株G31的赤藓糖醇滴度达到220.5 g/L，在5 L生物反应器中的生产率为1.8 g/L/h。该研究为基于生物传感器的超高通量筛选策略提供了宝贵的指导，并为粗甘油的可持续增值提供了新的范例。