Functional Identification of a Multi-product Sesquiterpene Synthase from Ganoderma sinense
10.13422/j.cnki.syfjx.20191216
- VernacularTitle: 一个紫芝多产物倍半萜合酶的功能鉴定
- Author:
Li-hua CHU
1
;
Li-zhi WANG
2
;
Shi-lin CHEN
3
;
Xin-yi ZENG
1
;
Jiang XU
3
;
Ying LI
1
;
Chao SUN
1
Author Information
1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
2. School of Chinese Material Medica, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
3. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
- Publication Type:Research Article
- Keywords:
Ganoderma sinense;
sesquiterpene synthase;
solid phage microextraction;
biosynthesis
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(12):151-157
- CountryChina
- Language:Chinese
-
Abstract:
Objective: Ganoderma sinense is one of the most famous medicinal fungi in China. Because it is a model medicinal fungus of basidiomycete,the functional identification of its sesquiterpene synthase is of great significance for the biosynthesis and regulation studies of fungal sesquiterpene. Method: A sesquiterpene synthase gene was discovered by mining the genome data of G. sinense. The sesquiterpene's conservative motifs was analyzed through multiple sequence alignment with two identified sesquiterpene synthases of G. sinense and three terpenoid synthases in the Nr database,which have the highest similarity to it. Subsequently,heterologous expression was observed in Escherichia coli,and protein expression was detected by SDS-PAGE. Volatile compounds were collected by solid phage microextraction (SPME) and detected by GC-MS. Result: The enzyme containing sesquiterpene conserved motifs DDXXDE and NSE/DTE were efficiently expressed in E. coli,and 11 sesquiterpenes were synthesized with endogenous FPP as the substrate. The product α-cadinol at 18.6 min was considered to be the main product,and previous studies showed a significant anticancer activity. According to the comparison with chemical standards,three products were identified as γ-muurlene,α-muurolene and δ-cadinene,respectively. Conclusion: The functional identification of multi-product sesquiterpene synthase from G. sinense can promote the study on the mechanism underlying its product diversity,and lay a foundation for the production of rare or novel sesquiterpenes by improving the catalytic activity of enzymes with enzyme engineering technology.
