Overexpression of Global Regulator LaeA Induced Secondary Metabolite Production in Aspergillus versicolor 0312
Overexpression of Global Regulator LaeA Induced Secondary Metabolite Production in Aspergillus versicolor 0312
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全局调节因子 LaeA 的过表达诱导杂色曲霉 0312 产生次级代谢产物
DOI：
10.25135/rnp.183.20.03.1593
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发表时间：
2020-11-01
影响因子：
1.9
通讯作者：
Wang, Weiguang
中科院分区：
文献类型：
--
作者：
He, Ting;Wang, Yudan;Wang, Weiguang
本实验研究了全局调控因子laeA对杂色曲霉次生代谢产物的影响。从杂色曲霉0312中克隆laeA基因并引入pSilent I过表达载体，通过PEG介导的原生质体转化将重组质粒导入杂色曲霉0312。然后，在代谢水平和基因水平对过表达菌株0312 - laeA进行筛选和验证。通过过表达菌株的液体发酵获得了一种新化合物杂色曲霉素A（1）以及四种已知化合物，乙酰阿朴阿拉诺丁（2）、乙酰阿拉诺丁（3）、邻苯二甲酸二异丁酯（4）和麦角甾醇（5）。利用包括高分辨电喷雾质谱（HRESIMS）、一维和二维核磁共振（1D - 和2D - NMR）数据在内的多种光谱技术阐明了新化合物的结构。化合物1对MOLT - 4细胞系显示出细胞毒性，半数抑制浓度（IC50）值为29.6 μM，而化合物2对MOLT - 4、MCF - 7和CaCo - 2细胞系显示出显著的细胞毒性，IC50值分别为7.8、19.9和15.9 μM。结果进一步证明laeA是一种全局调控因子，它可以上调和/或激活隐性基因簇以产生新的次生代谢产物。本研究为探索laeA基因对杂色曲霉次生代谢产物的作用机制提供了重要依据。
This experiment investigated the effect of the global regulator laeA on the secondary metabolites of Aspergillus versicolor. The laeA gene was cloned from Aspergillus versicolor 0312 to introduce pSilent I overexpression vector, and the recombinant plasmid was introduced into Aspergillus versicolor 0312 by PEG guided protoplast transformation. Then, the over-expressing strain 0312-laeA was screened and verified by metabolic level and gene level. A new compound versicolor A (1) was obtained by liquid fermentation of the over-expressing strain, together with four known compounds, acetylapoaranotin (2), acetylaranotin (3), diisobutyl phthalate (4) and ergosterol (5). The structure of the new compound was elucidated by using various spectroscopic techniques inclusive of HRESIMS, 1Dand 2D-NMR data. Compound 1 showed cytotoxicity against MOLT-4 cell lines with IC50 values of 29.6 μM, while 2 showed significant cytotoxicities against MOLT-4, MCF-7, and CaCo-2 cell lines with IC50 values of 7.8, 19.9, and 15.9 μM, respectively. The results further supported that laeA is a global regulator that could up-regulate and/or activate cryptic gene clusters to produce new secondary metabolites. This study provides an important basis for exploring the mechanism of laeA gene on the secondary metabolites of Aspergillus versicolor.