Optimization of the Biosynthesis of B-Ring Ortho-Hydroxy Lated Flavonoids Using the 4-Hydroxyphenylacetate 3-Hydroxylase Complex (HpaBC) of Escherichia coli.
Optimization of the Biosynthesis of B-Ring Ortho-Hydroxy Lated Flavonoids Using the 4-Hydroxyphenylacetate 3-Hydroxylase Complex (HpaBC) of Escherichia coli.
复制标题
使用大肠杆菌 4-羟基苯乙酸酯 3-羟化酶复合物 (HpaBC) 优化 B 环邻羟基延迟黄酮的生物合成
DOI：
10.3390/molecules26102919
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发表时间：
2021-05-14
期刊：
影响因子：
--
通讯作者：
Wang Y
中科院分区：
文献类型：
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作者：
Wang L;Ma X;Ruan H;Chen Y;Gao L;Lei T;Li Y;Gui L;Guo L;Xia T;Wang Y
黄酮类化合物是重要的植物代谢产物，具有广泛的生理和药理作用。由于其广泛的生物活性，如抗炎、抗氧化、抗衰老和抗癌等，已被广泛应用于食品、保健品和医药行业。在这里，选择羟化酶复合物HpaBC用于有效地在体内生产邻羟基化的黄酮类化合物。构建了几种HpaBC表达载体，并通过将柚皮素喂入载体携带菌株中，成功检测到相应的产物。然而，当HpaC与C末端的S-Tag连接时，酶活性受到显著影响。确定了最佳培养条件：底物浓度80 mg·L-1，诱导温度28 °C，M9培养基，IPTG诱导后底物延迟时间6 h。最后，以柚苷为底物，对圣草酚的转化率达到57.67 ± 3.36%。用同样的方法转化儿茶素和咖啡酸，最高转化率分别为35.2 ± 3.14%和32.93 ± 2.01%。本文首次证明了HpaBC对二氢山奈酚和山奈酚的催化活性。本研究证明了一种在细菌表达系统中体内有效合成B环二羟基黄酮类化合物，如儿茶素、黄烷醇、二氢黄酮醇和黄酮醇的可行方法。
Flavonoids are important plant metabolites that exhibit a wide range of physiological and pharmaceutical functions. Because of their wide biological activities, such as anti-inflammatory, antioxidant, antiaging and anticancer, they have been widely used in foods, nutraceutical and pharmaceuticals industries. Here, the hydroxylase complex HpaBC was selected for the efficient in vivo production of ortho-hydroxylated flavonoids. Several HpaBC expression vectors were constructed, and the corresponding products were successfully detected by feeding naringenin to vector-carrying strains. However, when HpaC was linked with an S-Tag on the C terminus, the enzyme activity was significantly affected. The optimal culture conditions were determined, including a substrate concentration of 80 mg·L−1, an induction temperature of 28 °C, an M9 medium, and a substrate delay time of 6 h after IPTG induction. Finally, the efficiency of eriodictyol conversion from P2&3-carrying strains fed naringin was up to 57.67 ± 3.36%. The same strategy was used to produce catechin and caffeic acid, and the highest conversion efficiencies were 35.2 ± 3.14 and 32.93 ± 2.01%, respectively. In this paper, the catalytic activity of HpaBC on dihydrokaempferol and kaempferol was demonstrated for the first time. This study demonstrates a feasible method for efficiently synthesizing in vivo B-ring dihydroxylated flavonoids, such as catechins, flavanols, dihydroflavonols and flavonols, in a bacterial expression system.