通过下调下游代谢通量，在 中工程顺式元件以有效积累附加值化合物角鲨烯。
Engineering of cis-Element in for Efficient Accumulation of Value-Added Compound Squalene via Downregulation of the Downstream Metabolic Flux.
机构信息
CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
出版信息
J Agric Food Chem. 2021 Oct 27;69(42):12474-12484. doi: 10.1021/acs.jafc.1c04978. Epub 2021 Oct 18.
Transcriptional downregulation is widely used for metabolic flux control. Here, , a cis-element of operator, was explored to engineer promoters of for downregulation. First, the promoter (P) and its enhanced variant P were engineered by insertion of into different sites, which resulted in decrease in both transcription and GFP fluorescence intensity to various degrees. Then, was applied to engineer the native and promoters due to their importance for accumulation of value-added intermediates squalene and lanosterol. Elevated squalene content (4.9-fold) or lanosterol content (4.8-fold) and 91 or 28% decrease in ergosterol content resulted from the engineered promoter P or P, respectively, indicating the validity of the engineered promoters in metabolic flux control. Furthermore, squalene production of 3.53 g/L from cane molasses, a cheap and bulk substrate, suggested the cost-effective and promising potential for squalene production.
转录下调被广泛用于代谢通量控制。在这里，我们探索了操纵子的顺式元件 ，以工程化用于下调的 启动子。首先，通过将 插入不同位置，工程改造了 启动子（P）及其增强变体 P，这导致 转录和 GFP 荧光强度都在不同程度上降低。然后，由于 对于积累有价值的中间产物角鲨烯和羊毛甾醇很重要，因此将 应用于工程化天然 启动子和 启动子。分别来自工程化启动子 P 或 P 的角鲨烯含量升高（4.9 倍）或羊毛甾醇含量升高（4.8 倍）以及甾醇含量降低 91%或 28%，表明这些工程化启动子在代谢通量控制中的有效性。此外，从廉价且大量的底物甘蔗蜜中生产 3.53 g/L 的角鲨烯表明，从甘蔗蜜中生产角鲨烯具有成本效益和广阔的应用前景。
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