Sun Rong, Liu Shan, Tang Zi-Zhong, Zheng Tian-Run, Wang Tao, Chen Hui, Li Cheng-Lei, Wu Qi
College of Life Science Sichuan Agricultural University Ya'an China.
College of Biological and Chemical Engineering Panzhihua University China.
FEBS Open Bio. 2017 Sep 6;7(10):1575-1585. doi: 10.1002/2211-5463.12299. eCollection 2017 Oct.
H.Lév. is a widely used medicinal herb in southwestern China. The main pharmacological components of are a class of oleanane-type pentacyclic triterpene glycosides known as conyzasaponins, which are thought to be synthesized from β-amyrin. However, no genes involved in the conyzasaponin pathway have previously been identified. Here, we identify an oxidosqualene cyclase (OSC), a β-amyrin synthase, which mediates cyclization of 2,3-oxidosqualene to yield β-amyrin. Ten OSC sequences were isolated from transcript tags. Phylogenetic analysis was used to select the tag Cb18076 as the putative β-amyrin synthase, named . The open reading frame of is 2286 bp and encodes 761 amino acids. Its mature protein contains the highly conserved motifs (QXXXGXW/DCTAE) of OSCs and (MWCYCR) of β-amyrin synthases. Transcription of was upregulated 4-24 h after treatment of the seedlings of the cultivar with methyl jasmonate. Furthermore, expression of in successfully yielded β-amyrin. The chemical structures and concentrations of β-amyrin were confirmed by GC-MS/MS. The target yeast ultimately produced 4.432 mg·L β-amyrin. Thus, CbβAS is an OSC involved in conyzasaponin biosynthesis.
H.Lév.是中国西南部广泛使用的一种药草。其主要药理成分是一类被称为豚草皂苷的齐墩果烷型五环三萜糖苷,据认为它们是由β-香树脂醇合成的。然而,此前尚未鉴定出参与豚草皂苷生物合成途径的基因。在此,我们鉴定出一种氧化鲨烯环化酶(OSC),即β-香树脂醇合酶,它介导2,3-氧化鲨烯环化生成β-香树脂醇。从转录本标签中分离出10个OSC序列。通过系统发育分析选择标签Cb18076作为推定的β-香树脂醇合酶,命名为CbβAS。CbβAS的开放阅读框为2286 bp,编码761个氨基酸。其成熟蛋白包含OSC的高度保守基序(QXXXGXW/DCTAE)和β-香树脂醇合酶的(MWCYCR)基序。用茉莉酸甲酯处理该品种幼苗后4 - 24小时,CbβAS的转录上调。此外,CbβAS在酵母中的表达成功产生了β-香树脂醇。通过气相色谱 - 串联质谱法确认了β-香树脂醇的化学结构和浓度。目标酵母最终产生了4.432 mg·L的β-香树脂醇。因此,CbβAS是一种参与豚草皂苷生物合成的OSC。