Molecular cloning and functional characterization of 2,3-oxidosqualene cyclases from Artemisia argyi

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS Protein expression and purification Pub Date : 2024-06-13 DOI:10.1016/j.pep.2024.106533

Yaman Chen , Ruoshi Huang , Jiabo Chen , Chumin Lin , Yuhong Wu , Jitong Chen , Qi Shen , Feng Wang , Lixin Duan , Honghua Cui

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Abstract

Artemisia argyi is a traditional medicinal and edible plant, generating various triterpenoids with pharmacological activities, such as anti-virus, anti-cancer, and anti-oxidant. The 2,3-oxidosqualene cyclase family of A. argyi offers novel insights into the triterpenoid pathway, which might contribute to the medicinal value of its tissue extracts. Nevertheless, the biosynthesis of active triterpenoids in Artemisia argyi is still uncertain. In this study, four putative OSC (2,3-oxidosqualene cyclase) genes (AaOSC1-4) were first isolated and identified from A. argyi. Through the yeast heterologous expression system, three AaOSCs were characterized for the biosynthesis of diverse triterpenoids including cycloartenol, β-amyrin, (3S,13R)-malabarica-14(27),17,21-trien-3β-ol, and dammara-20,24-dien-3β-ol. AaOSC1 was a multifunctional dammara-20,24-dien-3β-ol synthase, which yielded 8 different triterpenoids, including tricyclic, and tetracyclic products. AaOSC2 and AaOSC3 were cycloartenol, and β-amyrin synthases, respectively. As a result, these findings provide a deeper understanding of the biosynthesis pathway of triterpenes in A. argyi.

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蒿属植物中 2,3-氧化喹啉环化酶的分子克隆和功能鉴定。

阿尔基蒿是一种传统的药用和食用植物，可产生多种三萜类化合物，具有抗病毒、抗癌和抗氧化等药理活性。箭毒蒿的 2,3-氧化喹啉环化酶家族为三萜类化合物的途径提供了新的见解，这可能有助于提高其组织提取物的药用价值。然而，阿尔基蒿中活性三萜类化合物的生物合成仍然不确定。本研究首次从青蒿中分离并鉴定了四个推定的 OSC（2,3-oxidosqualene cyclase）基因（AaOSC1-4）。通过酵母异源表达系统，鉴定了三个 AaOSCs 在环木菠萝烯醇、β-amyrin、(3S,13R)-malabarica-14(27),17,21-trien-3β-ol 和 dammara-20,24-dien-3β-ol 等多种三萜类化合物的生物合成中的特性。AaOSC1 是一种多功能达玛拉-20,24-二烯-3β-醇合成酶，可产生 8 种不同的三萜类化合物，包括三环和四环产物。AaOSC2 和 AaOSC3 分别是环木菠萝烯醇合成酶和 β-amyrin 合成酶。因此，这些发现加深了人们对 A. argyi 三萜类化合物生物合成途径的了解。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.