Production of plant-derived anticancer precursor glucoraphanin in chromosomally engineered Escherichia coli.
Production of plant-derived anticancer precursor glucoraphanin in chromosomally engineered Escherichia coli.
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DOI：
10.1016/j.micres.2020.126484
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发表时间：
2020-05
影响因子：
6.7
通讯作者：
Han Yang;J. Qin;Xiuwen Wang;Bo Yu
中科院分区：
文献类型：
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作者：
Han Yang;J. Qin;Xiuwen Wang;Bo Yu
硫代葡萄糖苷是一种蛋氨酸衍生的硫代葡萄糖苷，具有广泛的生物活性和多种健康益处。萝卜硫苷在植物组织中含量低，提取成本高，限制了其生产。在异源微生物中进行代谢工程是实现有价值的天然产物的高效生产的有吸引力的方法。在这项研究中，一个微生物发酵生产萝卜硫苷的过程。该工程菌在大肠杆菌中稳定表达10种同种酶。colichromosome，包括9个异源基因，分别来自拟南芥和拟南芥，1个来自真菌粗糙脉孢菌（Neurospora crassa），该菌株能利用葡萄糖发酵生产硫代葡萄糖苷类化合物硫代葡萄糖苷，效价为0.675 μg/L。还研究了辅助因子补充剂和单个基因过表达对萝卜硫苷生产的影响。这项工作突出了通过微生物发酵过程而不是化学提取来提供专门植物硫代葡萄糖苷的可能性。此外，本研究中鉴定的限制性步骤酶UDP-葡萄糖-硫羟肟酸葡萄糖基转移酶也为进一步优化萝卜硫苷生产细胞工厂奠定了基础。
Glucoraphanin is a methionine-derived glucosinolate that imparts numerous health-benefits with broad bioactivity. Low amounts in plant tissues and high cost of extraction have limited the production of glucoraphanin. Metabolic engineering in heterologous microorganisms is an attractive approach to achieve efficient production of valuable natural products. In this study, a microbial fermentation process for glucoraphanin production was demonstrated. The engineered bacterial strain stably expressed 10 allogeneic enzymes inE. colichromosome, including nine heterologous genes fromArabidopsisandBrassicaand one from fungusNeurospora crassa, which could produce the specialized glucosinolate compound glucoraphanin with a titer of 0.675 μg/L by fermentation from glucose. The cofactor supplements and individual gene overexpression for glucoraphanin production were also investigated. This work highlights the possibility of supplying specialized plant glucosinolates by microbial fermentation process, instead of chemical extraction. Additionally, the limiting step enzyme, UDP-glucose-thiohydroximate glucosyltransferase, identified in this study also laid a foundation for further optimizing the glucoraphanin-producing cell factory.