Xu Tong Wang, Ting Ting Sun, Jian Sun, Shixin Wang and Li Zou*
Background: Sanghuangporus baumii is a traditional Chinese medicine with anti-cancer, anti-tumor, and anti-inflammatory effects. Triterpenoids are one of the main medicinal ingredients found in S. baumii. However, the dynamic changes of triterpenoids content and its molecular regulation mechanism are still unclear.
Objective: Some studies have shown that Lanosterol synthase (LS) is a key enzyme involved in the mevalonate pathway (MVA pathway) to produce lanosterol, which is a precursor for synthesizing S. baumii triterpenoids. Therefore, the study of LS gene and expression characteristics can provide clues for the further study of triterpenoids synthesis.
Methods: The PCR, RACE PCR, RT-PCR, seamless cloning and prokaryotic expression technology were used to research the gene characteristic and dynamic changes of LS transcription level.
Results: The S. baumii LS sequence included a 5’-untranslated region (129 bp), a 3’-untranslated region (87 bp), and an open reading frame (2,229 bp) encoding 734 amino acids. The S. baumii LS protein was expressed in E. coli BL21 (DE3). The transcription start site of the S. baumii LS promoter sequence ranged from 1 740 bp to 1790 bp. The LS promoter contained 12 CAAT-boxes, 5 ABREs, 6 G-Boxes, 6 CGTCA-motifs, and so on. The LS transcription levels were the highest on day 11 in mycelia (1.6-fold), and the triterpenoids content also gradually increased. The transcription levels began to decrease on day 13, but the triterpenoids content still increased.
Conclusion: The S. baumii LS was cloned and characterized to help to understand the mechanism of triterpenoids synthesis. In addition, we studied the relationship between LS transcription level and triterpenoid dynamic accumulation, and we found that they had a certain correlation.
Lanosterol synthase, Sanghuangporus baumii, Triterpenoids, Dynamic accumulation, Promoter, Protein expression
College of Forestry, Northeast Forestry University, Harbin 150040, Department of Food Engineering, Harbin University, Harbin 150086, College of Forestry, Northeast Forestry University, Harbin 150040, College of Forestry, Northeast Forestry University, Harbin 150040, College of Forestry, Northeast Forestry University, Harbin 150040