Overexpression of ribonucleotide reductase small subunit,RNRM,increases cordycepin biosynthesis in transformed Cordyceps militaris  

作　　者：ZHANG Han WANG Yu-Xian TONG Xin-Xin Yokoyama Wallace CAO Jing WANG Fang PENG Cheng GUO Jin-Lin 

机构地区：[1]Key Laboratory of Standardization of Chinese Medicine,Ministry of Education,Key Laboratoty 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

出　　处：《Chinese Journal of Natural Medicines》2020年第5期393-400,共8页中国天然药物（英文版）

基　　金：the Natural Sciences Foundation of China Science (Nos.81872959, 81373920,30801522);Sichuan Province Youth Innovation Team Fund (No.19CXTD0055);China Scholarship Fund (No. 201708570027)。

摘　　要：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 m RNA 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 wildtype 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.

关 键 词：Cordycepin biosynthesis Ribonucleotide reductases gene Agrobacterium tumefaciens-mediated transfection 

分 类 号：Q812[生物学—生物工程]