Phosphoenolpyruvate availability and the biosynthesis of shikimic acid
Phosphoenolpyruvate availability and the biosynthesis of shikimic acid
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DOI：
10.1021/bp025769p
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发表时间：
2003-05-01
影响因子：
2.9
通讯作者：
Frost, JW
中科院分区：
文献类型：
--
作者：
Chandran, SS;Yi, J;Frost, JW
在大肠杆菌K-12结构中，研究了在莽草酸生物合成过程中提高磷烯醇式丙酮酸利用率的影响，这些结构携带分别编码反馈不敏感的3-脱氧-D-阿拉伯-庚磺酸7-磷酸合成酶和转酮醇酶的质粒定位的aroF(FBR)和tktA插入片段。提高磷酸烯醇式丙酮酸利用率的策略是基于扩增表达大肠杆菌PPSA编码的磷酸烯醇式丙酮酸合成酶或异源表达运动发酵单胞菌GLF编码的葡萄糖促进剂。在缺乏磷酸烯醇式丙酮酸：碳水化合物磷酸转移酶系统的宿主中，利用同时表达运动链霉菌GLF编码的葡萄糖促进蛋白和运动链霉菌GLK编码的葡萄糖激酶的大肠杆菌SP1.1/pSC6.090B在1个L发酵罐中以葡萄糖为原料合成莽草酸的效价和产率最高。在添加酵母膏的10 L规模上，E.coliSP1.1/pSC6.090B以36%(摩尔/摩尔)的产率合成了87g/L的莽草酸，在指数生长期合成莽草酸的最高产率为5.2g/L/h。
The impact of increased availability of phosphoenolpyruvate during shikimic acid biosynthesis has been examined in Escherichia coli K-12 constructs carrying plasmid-localized aroF(FBR) and tktA inserts encoding, respectively, feedback-insensitive 3-deoxy-D-arabino-heptulosonic acid 7-phosphate synthase and transketolase. Strategies for increasing the availability of phosphoenolpyruvate were based on amplified expression of E. coli ppsA-encoded phosphoenolpyruvate synthase or heterologous expression of the Zymomonas mobilis glf-encoded glucose facilitator. The highest titers and yields of shikimic acid biosynthesized from glucose in 1 L fermentor runs were achieved using E. coli SP1.lpts/pSC6.090B, which expressed both Z. mobilis glf-encoded glucose facilitator protein and Z. mobilis glk-encoded glucose kinase in a host deficient in the phosphoenolpyruvate: carbohydrate phosphotransferase system. At 10 L scale with yeast extract supplementation, E. coli SP1.lpts/pSC6.090B synthesized 87 g/L of shikimic acid in 36% (mol/mol) yield with a maximum productivity of 5.2 g/L/h for shikimic acid synthesized during the exponential phase of growth.