Overexpression of ribonucleotide reductase small subunit, RNRM, increases cordycepin biosynthesis in transformed Cordyceps militaris.
10.1016/S1875-5364(20)30046-7
- Author:
Han ZHANG
1
;
Yu-Xian WANG
1
;
Xin-Xin TONG
1
;
Wallace YOKOYAMA
2
;
Jing CAO
1
;
Fang WANG
1
;
Cheng PENG
1
;
Jin-Lin GUO
3
Author Information
1. Key Laboratory of Standardization of Chinese Medicine, Ministry of Education; Key Laboratory of Systematic Research, Development and Utilization of Chinese Medicine, Resources Breeding Base of Co-founded by Sichuan Province and MOST, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.
2. USDA, ARS, Western Regional Research Center, Albany, CA 94710, USA.
3. Key Laboratory of Standardization of Chinese Medicine, Ministry of Education; Key Laboratory of Systematic Research, Development and Utilization of Chinese Medicine, Resources Breeding Base of Co-founded by Sichuan Province and MOST, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China. Electronic address: guo596@cdutcm.edu.cn.
- Publication Type:Journal Article
- Keywords:
Agrobacterium tumefaciens-mediated transfection;
Cordycepin biosynthesis;
Ribonucleotide reductases gene
- From:
Chinese Journal of Natural Medicines (English Ed.)
2020;18(5):393-400
- CountryChina
- Language:English
-
Abstract:
Cordycepin was the first adenosine analogue used as an anticancer and antiviral agent, which is extracted from Cordyceps militaris and hasn't been biosynthesized until now. This study was first conducted to verify the role of ribonucleotide reductases (RNRs, the two RNR subunits, RNRL and RNRM) in the biosynthesis of cordycepin by over expressing RNRs genes in transformed C. militaris. Quantitative real-time PCR (qRT-PCR) and western blotting results showed that the mRNA and protein levels of RNR subunit genes were significantly upregulated in transformant C. militaris strains compared to the control strain. The results of the HPLC assay indicated that the cordycepin was significantly higher in the C. militaris transformants carrying RNRM than in the wild-type strain, whereas the RNRML was preferentially downregulated. For the C. militaris transformant carrying RNRL, the content of cordycepin wasn't remarkably changed. Furthermore, we revealed that inhibiting RNRs with Triapine (3-AP) almost abrogated the upregulation of cordycepin. Therefore, our results suggested that RNRM can probably directly participate in cordycepin biosynthesis by hydrolyzing adenosine, which is useful for improving cordycepin synthesis and helps to satisfy the commercial demand of cordycepin in the field of medicine.
