Cloning and bioinformatics analysis of β-amyrin synthase in Dipsacus asper.
10.19540/j.cnki.cjcmm.20220514.102
- Author:
Wei-Lin YAO
1
;
Jie PAN
1
;
Teng-Fei NIU
1
;
Xiao-Lin YANG
2
;
Shu-Juan ZHAO
2
;
Zheng-Tao WANG
2
;
Ru-Feng WANG
2
Author Information
1. Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine Shanghai 201203, China the SATCM Key Laboratory for New Resources & Quality Evaluation of Chinese Medicine, Shanghai University of Traditional Chinese Medicine Shanghai 201203, China.
2. Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine Shanghai 201203, China the SATCM Key Laboratory for New Resources & Quality Evaluation of Chinese Medicine, Shanghai University of Traditional Chinese Medicine Shanghai 201203, China Shanghai R&D Center for Standardization of Traditional Chinese Medicines Shanghai 201203, China the MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai University of Traditional Chinese Medicine Shanghai 201203, China.
- Publication Type:Journal Article
- Keywords:
Dipsacus asper;
bioinformatics;
gene cloning;
triterpenoid saponins;
β-amyrin synthase
- MeSH:
Cloning, Molecular;
Computational Biology;
Dipsacaceae/chemistry*;
Intramolecular Transferases;
Protein Sorting Signals;
Saponins/chemistry*;
Triterpenes/chemistry*
- From:
China Journal of Chinese Materia Medica
2022;47(17):4593-4599
- CountryChina
- Language:Chinese
-
Abstract:
Dipsaci Radix is one of the commonly used Chinese medicinal materials in China, with a long history. It has the medicinal activities of nourishing liver and kidney, recovering from broken sinews, and treating bone fracture. Triterpenoid saponins are the main functional ingredients of Dipsacus asper. β-Amyrin synthases(β-AS) as a superfamily of oxidosqualene cyclases(OSCs) can catalyze the construction of the skeleton structure of oleanane-type triterpenoid saponins. There are only a few studies about the β-AS in D. asper, and the catalytic mechanism of this enzyme remains to be explored. To enrich the information of β-AS, according to the transcriptome sequencing results, we cloned DaWβ-AS gene from D. asper into a specific vector for heterologous expression in Escherichia coli. In the meantime, real-time PCR was performed to analyze the relative expression of DaWβ-AS in four different tissues of D. asper. The results of RT-qPCR showed DaWβ-AS had the highest expression level in leaves. Bioinformatics results indicated that DaWβ-AS had a conserved domain of PLN03012 superfamily, belonging to the cl31551 superfamily. There was no transmembrane domain or signal peptide in DaWβ-AS. This study provides a scientific basis for revealing the biological pathways of triterpenoid saponins in D. asper, which will facilitate the biosynthesis of the associated saponins and afford reference for the cultivation and development of high-quality resources of D. asper.
