Biosynthetic Pathway Analysis for Improving the Cordycepin and Cordycepic Acid Production in Hirsutella sinensis
Biosynthetic Pathway Analysis for Improving the Cordycepin and Cordycepic Acid Production in Hirsutella sinensis
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DOI：
10.1007/s12010-016-2020-0
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发表时间：
2016-06-01
影响因子：
3
通讯作者：
Zheng, Yu-Guo
中科院分区：
文献类型：
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作者：
Lin, Shan;Liu, Zhi-Qiang;Zheng, Yu-Guo
中华被毛菌被认为是冬虫夏草唯一正确的无性型。为了提高冬虫夏草中虫草素和虫草酸的产量，预测了虫草素和虫草酸的生物合成途径，并分别通过克隆和表达这些途径所涉及的基因来验证。然后，通过实时PCR证明参与虫草素生物合成途径的5'-核苷酸酶、己糖激酶和参与虫草酸生物合成途径的葡萄糖磷酸异构酶在相应的生物合成途径中发挥重要作用，并分别显着上调15.03倍、5.27倍和3.94倍。此外，还对中华线虫的代谢进行了调控。正如预期的那样，当 5'-核苷酸酶的附加底物为 4 mg/mL 时，虫草素产量达到 1.09 mg/g，与对照相比增加了 201.1%。同样，添加己糖激酶或葡萄糖磷酸异构酶底物，虫草酸产量分别提高了26.6%和23.4%，分别提高了77.3%和55.1%。迄今为止，该研究首次基于生物合成途径分析通过代谢调控来提高虫草素和虫草酸产量，证明代谢调控是提高冬虫夏草深层培养虫草素和虫草酸产量的简单有效的方法。
Hirsutella sinensis is considered as the only correct anamorph of Ophiocordyceps sinensis. To improve cordycepin and cordycepic acid production in H. sinensis, the biosynthetic pathways of cordycepin and cordycepic acid were predicted, and verified by cloning and expressing genes involved in these pathways, respectively. Then, 5'-nucleotidase participating in biosynthetic pathway of cordycepin, hexokinase, and glucose phosphate isomerase involved in biosynthetic pathway of cordycepic acid, were demonstrated playing important roles in the corresponding biosynthetic pathway by real-time PCR, accompanying with significantly up-regulated 15.03-, 5.27-, and 3.94-fold, respectively. Moreover, the metabolic regulation of H. sinensis was performed. As expected, cordycepin production reached 1.09 mg/g when additional substrate of 5'-nucleotidase was 4 mg/mL, resulting in an increase of 201.1 % compared with the control. In the same way, cordycepic acid production reached 26.6 and 23.4 % by adding substrate of hexokinase or glucose phosphate isomerase, leading to a rise of 77.3 and 55.1 %, respectively. To date, this is the first time to improve cordycepin and cordycepic acid production through metabolic regulation based on biosynthetic pathway analysis, and metabolic regulation is proved as a simple and effective way to enhance the output of cordycepin and cordycepic acid in submerged cultivation of H. sinensis.