Establishing a new methodology for genome mining and biosynthesis of natural products through fungal molecular genomics
Establishing a new methodology for genome mining and biosynthesis of natural products through fungal molecular genomics
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DOI：
10.1055/s-0032-1320342
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发表时间：
2012-07
期刊：
影响因子：
2.7
通讯作者：
H. Noguchi
中科院分区：
文献类型：
--
作者：
H. Noguchi
聚酮化合物（PKs）和非核糖体肽（NRP）已从链霉菌和许多其他来源的生物体中分离出来。近年来，通过真菌基因组测序发现了编码PK代谢酶（PKSs）和NRP合成酶（NRPSs）的基因簇。虽然在单个真菌基因组中鉴定出平均50个基因簇，但可以从在典型生长条件下生长的真菌培养物中分离出较少的真菌PK和NRP产物。为了克服这些障碍，我们研究了四种真菌，烟曲霉，A. Flavus、黄毛菊A.和球毛壳菌，使用上述真菌分子遗传学。到目前为止，我们已经成功地分离了七个新的PK和NRP化合物。随后，我们使用我们的重组克隆为基础的酵母表达系统，快速，有效地重建这些生物合成基因簇。我们的初步结果清楚地表明成功表达了7个C。globosum PKS基因簇在酿酒酵母，其中三个导致新的天然产物的身份已被表征光谱的生产。我们的方法将有助于分离新的天然产物和合理地工程化的生物合成途径，以生产具有可比的，如果不是更有效的生物活性的类似物。
Polyketides (PKs) and nonribosomal peptides (NRPs) have been isolated from Streptomyces and many other source organisms. In recent years, gene clusters encoding PK synthases (PKSs) and NRP synthetases (NRPSs) have been discovered through fungal genome sequencing. While on average 50 gene clusters are identified in a single fungal genome, fewer fungal PK and NRP products can be isolated from a fungal culture grown under a typical growth condition. Thus, simple artificial reactivation of the cryptic gene cluster may be insufficient for an efficient natural product biosynthesis.To circumvent these obstacles, we examined upregulation of 60 gene clusters encoded in chromosomal DNA of four fungal species, Aspergillus fumigatus, A. flavus, A. oryzae and Chaetomium globosum, using the aforementioned fungal molecular genetics. Thus far, we have isolated seven new PK and NRP compounds successfully. Subsequently, we used our recombination cloning-based yeast expression system to reconstitute these biosynthetic gene clusters quickly and efficiently. Our preliminary results clearly demonstrate successful expression of seven C. globosum PKS gene clusters in Saccharomyces cerevisiae, three of which led to the production of new natural products whose identities have been characterized spectroscopically. Our methodology will facilitate the efforts in isolating novel natural products and rationally engineering in the biosynthetic pathways for production of analogs possessing comparable if not more potent bioactivity.