Xu Yayun, Zhang Sixuan, Meng Fanqi, Liang Wenjing, Peng Yunliang, Zhu Butuo, Niu Lili, Wang Chunling, Li Caili, Lu Shanfa
State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-Di Herbs, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.
Key Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.
Int J Mol Sci. 2025 May 1;26(9):4314. doi: 10.3390/ijms26094314.
Bunge has been widely used as traditional Chinese medicine materials. However, its utility faces a great challenge due to the presence of aristolochic acids (AAs), a class of benzylisoquinoline alkaloid (BIA) derivatives. The first step in BIA skeleton formation is catalysis by norcoclaurine synthase (NCS). To gain knowledge of BIA and AA biosynthesis in , genome-wide characterizations of genes were carried out. This resulted in the identification of 15 , namely, . The AcNCS1-AcNCS8 proteins contained one catalytic domain, whereas the AcNCS9-AcNCS15 proteins had two. Phylogenetic analysis shows that AcNCS proteins can be classified into two clades. Gene expression analysis shows that five , including , , , , and , exhibited relatively high expression in roots and flowers, where norcoclaurine accumulated. An enzyme catalytic activity assay shows that all five of the AcNCSs can catalyze norcoclaurine formation with AcNCS14 and AcNCS15, exhibiting higher catalytic efficiency. Precolumn derivatization analysis shows that the formed norcoclaurine included ()- and ()-norcoclaurine, with more ()-configuration. The results provide useful information for further understanding BIA and AA biosynthesis in and for AA elimination and bioactive compound improvement in AA-containing medicinal materials.
防己已被广泛用作中药材。然而，由于马兜铃酸（AAs）的存在，其应用面临巨大挑战，马兜铃酸是一类苄基异喹啉生物碱（BIA）衍生物。BIA骨架形成的第一步是由去甲乌药碱合酶（NCS）催化。为了了解防己中BIA和AA的生物合成，对相关基因进行了全基因组表征。这导致鉴定出15个NCS基因，即AcNCS1 - AcNCS15。AcNCS1 - AcNCS8蛋白含有一个催化结构域，而AcNCS9 - AcNCS15蛋白有两个。系统发育分析表明，AcNCS蛋白可分为两个进化枝。基因表达分析表明，五个NCS基因，包括AcNCS1、AcNCS2、AcNCS3、AcNCS14和AcNCS15，在去甲乌药碱积累的根和花中表现出相对较高的表达。酶催化活性测定表明，所有五个AcNCS都能催化去甲乌药碱的形成，其中AcNCS14和AcNCS15表现出更高的催化效率。柱前衍生化分析表明，形成的去甲乌药碱包括（+）-和（-）-去甲乌药碱，其中（+）-构型更多。这些结果为进一步了解防己中BIA和AA的生物合成以及含AA药材中AA的去除和生物活性化合物的改良提供了有用信息。
