Cloning and Expression Analysis of CaCYP716E41 Gene from Centella asiatica
Men Yuchen
Chang Wanqi
Cai Qihang
Li Yulun
Sun Zhenghai
Li Liping
Abstract：Oleanolic acid 6-beta-oxidase(CYP716E41)is a backbone-modifying enzyme of triterpe-noids,and regulates the composition of triterpenoids with different structures.Oleanolic acid(OA)is one of the important triterpenoids with a variety of biological activities.In this study,the CaCYP716E41 gene was cloned from Centella asiatica leaf tissue by RT-PCR and was analyzed by bioinformatics analysis.The results showed that the CaCYP716E41 gene had a CDS sequence of 1 434 bp encoding 477 amino acids,and the Ca-CYP716E41 protein had a molecular weight of 53.88 kDa and an isoelectric point of 8.74,classified as a hy-drophilic stabilized protein.Protein structure analysis indicated that CaCYP716E41 protein had multiple heme binding sites,categorized as a member of the cytochrome P450 superfamily,and contained a transmembrane structure and had 44 phosphorylation sites.The results of the phylogenetic tree based on the CYP716E41 pro-tein sequence showed that CaCYP716E41 protein was the most closely related to the CYP716E41 protein of Ip-omoea batatas and Buddleja alternifolia.Tissue expression analysis demonstrated that CaCYP716E41 was ex-pressed in roots,stems and leaves,with varying relative expression levels among different tissues,exhibiting the highest expression in leaves.Meanwhile,when nitrogen,phosphorus and potassium fertilizers were applied to Centella asiatica respectively to study the regulation of different fertilizers on CaCYP716E41 expression,the results showed that the relative expression of CaCYP716E41 gene in roots,stems and leaves were elevated to different degrees by fertilization,and the expression of this gene was the highest in the stems of Centella asiati-ca after potassium fertilizer treatment,which was about 9.3 times of that of CK.It was hypothesized that nitro-gen,phosphorus and potassium fertilizers could promote the synthesis of oleanolic acid 6-beta-oxidase.The re-sults of this study could provide a reference for in-depth investigation of the function of CaCYP716E41 gene and lay the foundation for further research on biosynthetic pathway of triterpenoids in Centella asiatica.
Keywords：Centella asiaticaTriterpenoidOleanolic acid 6-beta-oxidaseGene cloningBioinformatics analysisExpression analysis
Publication Date：2026-02-28
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：8( 26-33 )