Cloning and characterization of chalcone synthase and chalcone isomerase genes in Arisaema heterophyllum.
10.19540/j.cnki.cjcmm.20190130.001
- Author:
Sheng-Xiang ZHANG
1
;
Yuan-Yuan SHI
1
;
Chen-Kai WANG
1
;
De-Rui ZHAO
1
;
Qing-Shan YANG
2
;
Ke-Long MA
3
;
Jia-Wen WU
4
Author Information
1. Graduate School,Anhui University of Chinese Medicine Hefei 230012,China Key Laboratory of Xin'an Medicine,Ministry of Education,Anhui University of Chinese Medicine Hefei 230038,China.
2. College of Pharmacy,Anhui University of Chinese Medicine Hefei 230012,China Synergetic Innovation Center of Anhui Authentic Chinese Medicine Quality Improvement Hefei 230012,China Anhui Academy of Chinese Medicine Hefei 230012,China.
3. Clinical College of Integrated Traditional Chinese and Western Medicine,Anhui University of Chinese Medicine Hefei 230012,China.
4. Key Laboratory of Xin'an Medicine,Ministry of Education,Anhui University of Chinese Medicine Hefei 230038,China Synergetic Innovation Center of Anhui Authentic Chinese Medicine Quality Improvement Hefei 230012,China Anhui Academy of Chinese Medicine Hefei 230012,China.
- Publication Type:Journal Article
- Keywords:
Arisaema heterophyllum;
bioinformatics analysis;
chalcone isomerase;
chalcone synthase;
gene cloning
- MeSH:
Acyltransferases;
chemistry;
genetics;
Arisaema;
enzymology;
genetics;
Cloning, Molecular;
Intramolecular Lyases;
chemistry;
genetics;
Plant Proteins;
chemistry;
genetics
- From:
China Journal of Chinese Materia Medica
2019;44(9):1799-1807
- CountryChina
- Language:Chinese
-
Abstract:
Chalcone synthase( CHS) and chalcone isomerase( CHI) are key enzymes in the biosynthesis pathway of flavonoids. In this study,unigenes for CHS and CHI were screened from the transcriptome database of Arisaema heterophyllum. The open reading frame( ORFs) of chalcone synthase( Ah CHS) and chalcone isomerase( Ah CHI) were cloned from the plant by RT-PCR. The physicochemical properties,expression and structure characteristics of the encoded proteins Ah CHS and Ah CHI were analyzed. The ORFs of Ah CHS and Ah CHI were 1 176,630 bp in length and encoded 392,209 amino acids,respectively. Ah CHS functioned as a symmetric homodimer. The N-terminal helix of one monomer entwined with the corresponding helix of another monomer. Each CHS monomer consisted of two structural domains. In particular,four conserved residues define the active site. The tertiary structure of Ah CHI revealed a novel open-faced β-sandwich fold. A large β-sheet( β4-β11) and a layer of α-helices( α1-α7) comprised the core structure. The residues spanning β4,β5,α4,and α6 in the three-dimensional structure were conserved among CHIs from different species. Notably,these structural elements formed the active site on the protein surface,and the topology of the active-site cleft defined the stereochemistry of the cyclization reaction. The homology comparison showed that Ah CHS had the highest similarity to the CHS of Anthurium andraeanum,while Ah CHI had the highest similarity to the CHI of Paeonia delavayi. This study provided the basis for the functional study of Ah CHS and Ah CHI and the further study on plant flavonoid biosynthesis pathway.
