Pathway optimization by re-design of untranslated regions for L-tyrosine production in Escherichia coli.
Pathway optimization by re-design of untranslated regions for L-tyrosine production in Escherichia coli.
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DOI：
10.1038/srep13853
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发表时间：
2015-09-08
影响因子：
4.6
通讯作者：
Jung GY
中科院分区：
文献类型：
--
作者：
Kim SC;Min BE;Hwang HG;Seo SW;Jung GY
L-酪氨酸是食品、制药、化工和化妆品行业中一种重要的商业化合物。虽然已经进行了几次尝试来提高L-酪氨酸的生产，但仍然需要实现磷酸烯醇丙酮酸（PEP）节点周围的抑制水平表达控制和碳通量再平衡以优化该途径。在这里，我们展示了通过改变大肠杆菌中L-酪氨酸生产的基因表达水平来优化途径。为了优化L-酪氨酸生物合成途径，为每个感兴趣的基因引入合成的组成型启动子和合成的5′-非翻译区（5′-UTR），以允许在转录和翻译水平上进行控制。通过使用UTR Designer控制PEP合成酶的表达水平来实现碳通量再平衡。工程菌的L-酪氨酸生产能力。大肠杆菌菌株通过途径优化，L-酪氨酸效价提高到3.0 g/L，产率提高到0.0354 g L-酪氨酸/h/g DCW，葡萄糖产量提高到0.102 g L-酪氨酸/g DCW。因此，这项工作表明，通过5′-UTR重新设计的途径优化是开发高效L-酪氨酸生产细菌的有效策略。
L-tyrosine is a commercially important compound in the food, pharmaceutical, chemical, and cosmetic industries. Although several attempts have been made to improve L-tyrosine production, translation-level expression control and carbon flux rebalancing around phosphoenolpyruvate (PEP) node still remain to be achieved for optimizing the pathway. Here, we demonstrate pathway optimization by altering gene expression levels for L-tyrosine production in Escherichia coli. To optimize the L-tyrosine biosynthetic pathway, a synthetic constitutive promoter and a synthetic 5′-untranslated region (5′-UTR) were introduced for each gene of interest to allow for control at both transcription and translation levels. Carbon flux rebalancing was achieved by controlling the expression level of PEP synthetase using UTR Designer. The L-tyrosine productivity of the engineered E. coli strain was increased through pathway optimization resulting in 3.0 g/L of L-tyrosine titer, 0.0354 g L-tyrosine/h/g DCW of productivity, and 0.102 g L-tyrosine/g glucose yield. Thus, this work demonstrates that pathway optimization by 5′-UTR redesign is an effective strategy for the development of efficient L-tyrosine-producing bacteria.