主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

JOURNAL OF BEIJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE ›› 2019, Vol. 42 ›› Issue (1): 69-76.doi: 10.3969/j.issn.1006-2157.2019.01.011

• Chinese Medicinal Resources • Previous Articles     Next Articles

Influence of KSL gene polymorphism of Danshen on enzyme-catalyzed reaction of its codases

Jin Xiaoyan1, Qiu Minyi1, Shao Yuanyang1, Wang Juan2, Li Geng3, Wang Xueyong1#   

  1. 1 School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China;
    2 Hangzhou TigerMed Technology, Zhejiang 310053, China;
    3 China-Japan Friendship Hospital, Beijing 100029, China
  • Received:2018-07-13 Online:2019-01-30 Published:2019-03-01
  • Contact: Wang Xueyong, male, PhD., Professor, Doctoral Supervisor. Research direction: medicinal plants and molecular pharmacognosy. E-mail: wxyph.d@163.com
  • Supported by:
    National Natural Science Foundation of China (No. 81773841), Central-level Major Increase, Decrease and Expenditures Project (No. 2060302).

Abstract: Objective To analyze the variation types kaunene synthase-like (KSL) gene in Danshen (Salvia Root, Radix Salviae Miltiorrhizae) from different producing areas in China, and investigate primarily the influence of KSL gene polymorphism on enzyme-catalyzed reaction of its codases. Methods Specific primers were designed, and full-length KSL genes of Danshen from different producing areas (Henan, Shandong, Sichuan, Hubei, Shanxi, Neimonggu, Hebei) in China were homologously cloned. The variation types were compared and mutation samples were selected to establish a prokaryotic expression system and inducible expression in vitro. The enzyme-catalyzed products of KSL gene were detected by using gas chromatography-mass spectrometer (GS-MS), and influence of KSL gene polymorphism of Danshen on protease catalytic efficiency was further analyzed. Results The full-length KSL gene was 1 788 bp, and there were totally 595 amino acid residues encoded. The sequencing results showed that there were polymorphisms in KSL gene of Danshen from different producing areas in China, which could cause synonymous and non-synonymous mutations. The catalytic efficiency of different genotypes of KSL was significantly different, which was the highest in Shanxi and the lowest in Inner Mongolia. The KSL gene containing A-type variation mutated at 1 047 bp in Danshen from 4 producing areas including Shanxi, Hubei, Shandong, and Sichuan, and the catalytic efficiency of which was significantly higher than those containing no A-type variation in Danshen from 4 producing areas including Henan, Hebei and Inner Mongolia. Conclusion KSL genes have gene polymorphisms indeed in Danshen from different producing areas. The non-synonymous mutations induced by polymorphisms can further lead to changes of amino acid sequences and finally resulting in difference in the catalytic efficiency of encoded enzymes. The polymorphisms of key enzyme genes in the synthesis pathway of tanshinones can be taken as a starting point to provide further reference for revealing the accumulation mechanism of tanshinones and breeding the excellent germplasm of Danshen.

Key words: Danshen (Salvia Root, Radix Salviae Miltiorrhizae), kaunene synthase-like gene, key enzyme, gene polymorphism, catalyzed efficiency

CLC Number: 

  • R282.71