Liu Tingxia, Zhang Wanran, Wang Sijia, Tian Ya, Wang Yifan, Gao Ranran, Chen Shilin, Sun Wei, Ma Wei, Xu Zhichao
College of Pharmaceutical Sciences, Heilongjiang University of Chinese Medicine, Harbin 150040, China.
Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Chin Herb Med. 2024 Nov 9;17(1):178-188. doi: 10.1016/j.chmed.2024.11.003. eCollection 2025 Jan.
Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in .
The transcriptome and metabolome data from 18 different tissues of were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in , a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity.
The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into ()-norcoclaurine.
Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.
苄基异喹啉生物碱（BIAs）具有药理功能和临床用途。BIAs主要分布在毛茛目以及无患子目某些属的植物物种中。BIAs的生物合成已在毛茛目物种中得到深入研究。然而，BIAs在（此处原文缺失相关植物名称）中的积累机制在很大程度上尚不清楚。本研究的目的是揭示（此处原文缺失相关植物名称）中BIAs的生物合成途径。
对（此处原文缺失相关植物名称）18个不同组织的转录组和代谢组数据进行了细致测序，随后进行了全面分析。加权基因共表达网络分析（WGCNA），一种有助于构建和后续分析共表达网络的强大系统生物学方法，被用于鉴定参与BIAs生物合成的候选基因。在此之后，含有候选基因的重组质粒在大肠杆菌（E. coli）中表达，大肠杆菌是一种广泛使用的原核表达系统。这项基因工程工作的目的是在细菌中表达候选基因，从而能够评估产生的酶活性。
旁系同源基因每个同义位点的同义替换表明至少发生了一次全基因组复制事件。提出了（此处原文缺失相关植物名称）潜在的BIAs生物合成途径，选择了两个PR10/Bet v1成员、14个细胞色素P450（CYP450s）和33个甲基转移酶与BIAs生物合成相关。一个PR10/Bet v1被鉴定为去甲乌药碱合酶，它可以催化多巴胺和4-羟基苯乙醛生成（）-去甲乌药碱。
我们的研究为非毛茛目物种中BIAs的生物合成和进化提供了重要见解。
