Improving key enzyme activity in phenylpropanoid pathway with a designed biosensor
Improving key enzyme activity in phenylpropanoid pathway with a designed biosensor
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利用设计的生物传感器提高苯丙素途径中的关键酶活性
DOI：
10.1016/j.ymben.2017.01.006
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发表时间：
2017
影响因子：
8.4
通讯作者：
Tang Shuang Yan
中科院分区：
文献类型：
--
作者：
Xiong D;an;Lu Shikun;Wu Jieyuan;Liang Chaoning;Wang Wei;Wang Wenzhao;Jin Jian Ming;Tang Shuang Yan
对香豆酸：辅酶A连接酶（p-Coumarate：CoA ligase，4CL）是苯丙素类化合物合成途径中的一种关键酶，是合成多种天然产物的关键酶。为了筛选具有提高活性的4CL，通过工程化TtgR调节蛋白设计了生物合成途径涉及4CL的白藜芦醇生物传感器。该生物传感器具有良好的特异性和鲁棒性，可以快速，灵敏地选择白藜芦醇超生产者。从构建的白藜芦醇生物合成途径4CL突变体库中筛选出一个活性提高的4CL突变体。当引入相应的生物合成途径时，该突变体不仅导致白藜芦醇的产量增加，而且还导致类黄酮柚皮素的产量增加。这些研究结果表明，在重要的生物合成途径中的关键酶的活性与援助的途径产品的设计生物传感器的可行性。
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 inEscherichia 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.