| 论文题目: | Improving key enzyme activity in phenylpropanoid pathway with a designed biosensor |
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| 作者: | Xiong Dandan , Lu Shikun , Wu Jieyuan , Liang Chaoning , Wang Wei , Wang Wenzhao , Jin Jian-Ming*, and Tang Shuang-Yan*. |
| 联系作者: | |
| 刊物名称: | Metabolic Engineering |
| 期: | |
| 卷: | 40 |
| 页: | 115-123 |
| 年份: | 2017 |
| 影响因子: | 8.322 |
| 论文下载: | 下载地址 |
| 摘要: | Overexpressing key enzymes of biosynthetic pathways for overproduction of value-added products usually imposes metabolic burdens on cells, which can be circumvented by improving the key enzyme activities. p-Coumarate: CoA ligase (4CL) is a critical enzyme in the phenylpropanoid pathway that synthesizes various natural products. To screen for 4CL with improved activity, a biosensor of resveratrol whose biosynthetic pathway involves 4CL was designed by engineering the TtgR regulatory protein. The biosensor exhibited good specificity and robustness, allowing rapid and sensitive selection of resveratrol hyper-producers. A 4CL variant with improved activity was selected from a 4CL mutagenesis library constructed in the resveratrol biosynthetic pathway in Escherichia coli. This mutant led to increased production of not only resveratrol but also the flavonoid naringenin, when introduced in their corresponding biosynthetic pathways. These findings demonstrate the feasibility of improving key enzyme activities in important biosynthetic pathways with the aid of designed biosensors of pathway products. |
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