Phosphoenolpyruvate Availability and the Biosynthesis of Shikimic Acid

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• 作者：Sunil S. Chandran ; Jian Yi ; K. M. Draths ; Ralph von Daeniken ; Wolfgang Weber ; and J. W. Frost
• 刊名：Biotechnology Progress
• 出版年：2003
• 出版时间：June 2003
• 年：2003
• 卷：19
• 期：3
• 页码：808 - 814
• 全文大小：172K
• 年卷期：v.19,no.3(June 2003)
• ISSN：1520-6033

文摘

The impact of increased availability of phosphoenolpyruvate during shikimic acidbiosynthesis 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 forincreasing the availability of phosphoenolpyruvate were based on amplified expressionof E. coli ppsA-encoded phosphoenolpyruvate synthase or heterologous expression ofthe Zymomonas mobilis glf-encoded glucose facilitator. The highest titers and yieldsof shikimic acid biosynthesized from glucose in 1 L fermentor runs were achieved usingE. coli SP1.lpts/pSC6.090B, which expressed both Z. mobilis glf-encoded glucosefacilitator protein and Z. mobilis glk-encoded glucose kinase in a host deficient in thephosphoenolpyruvate:carbohydrate phosphotransferase system. At 10 L scale withyeast extract supplementation, E. coli SP1.lpts/pSC6.090B synthesized 87 g/L ofshikimic acid in 36% (mol/mol) yield with a maximum productivity of 5.2 g/L/h forshikimic acid synthesized during the exponential phase of growth.