以葡萄糖为唯一碳源生产木糖的大肠杆菌代谢工程,ACS Synthetic Biology

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以葡萄糖为唯一碳源生产木糖的大肠杆菌代谢工程
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2021-08-19 , DOI: 10.1021/acssynbio.1c00184
Wencheng Yin ₁ , Yujin Cao ₁ , Miaomiao Jin ₁ , Mo Xian ₁ , Wei Liu ₁

Affiliation

木糖是合成许多重要平台化合物的原料。目前，木糖通过化学萃取进行商业化生产。然而，木糖的提取仍存在一些瓶颈，包括操作工艺复杂、化学物质引入等，导致木糖和木糖下游化合物合成成本高昂。目前木糖的市场价格是葡萄糖的8倍，因此以低成本的葡萄糖为底物生产木糖下游化合物理论上可以降低70%的成本。在这里，我们设计了一条在大肠杆菌中从葡萄糖生物合成木糖的途径。该生物合成途径是通过过表达五个基因实现的，即zwf、pgl、gnd、rpe和xylA，同时将天然木酮糖激酶基因xylB替换为来自枯草芽孢杆菌的araL，其显示出对d-木酮糖5-磷酸的磷酸酶活性。通过优化代谢途径，木糖产量提高到3.3 g/L。此外，通过引入xyl1成功合成了木糖醇。基因，这表明葡萄糖生物合成木糖的途径普遍适用于木糖下游化合物的合成。这是首次以葡萄糖为底物合成木糖及其下游化合物的研究，不仅降低了原料成本，而且缓解了碳分解代谢物抑制（CCR），为合成木糖下游化合物提供了新思路。木糖。

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Metabolic Engineering of E. coli for Xylose Production from Glucose as the Sole Carbon Source

Xylose is the raw material for the synthesis of many important platform compounds. At present, xylose is commercially produced by chemical extraction. However, there are still some bottlenecks in the extraction of xylose, including complicated operation processes and the chemical substances introduced, leading to the high cost of xylose and of synthesizing the downstream compounds of xylose. The current market price of xylose is 8× that of glucose, so using low-cost glucose as the substrate to produce the downstream compounds of xylose can theoretically reduce the cost by 70%. Here, we designed a pathway for the biosynthesis of xylose from glucose in Escherichia coli. This biosynthetic pathway was achieved by overexpressing five genes, namely, zwf, pgl, gnd, rpe, and xylA, while replacing the native xylulose kinase gene xylB with araL from B. subtilis, which displays phosphatase activity toward d-xylulose 5-phosphate. The yield of xylose was increased to 3.3 g/L by optimizing the metabolic pathway. Furthermore, xylitol was successfully synthesized by introducing the xyl1 gene, which suggested that the biosynthetic pathway of xylose from glucose is universally applicable for the synthesis of xylose downstream compounds. This is the first study to synthesize xylose and its downstream compounds by using glucose as a substrate, which not only reduces the cost of raw materials, but also alleviates carbon catabolite repression (CCR), providing a new idea for the synthesis of downstream compounds of xylose.

更新日期：2021-09-17

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