Fujita Masaki J, Kimura Nobutada, Sakai Atsushi, Ichikawa Yoichi, Hanyu Tomohiro, Otsuka Masami
Priority Organization for Innovation and Excellence, Kumamoto University, Japan.
Biosci Biotechnol Biochem. 2011;75(12):2283-7. doi: 10.1271/bbb.110379. Epub 2011 Dec 7.
A biosynthetic gene cluster of siderophore consisting of five open reading frames (ORFs) was cloned by functional screening of a metagenomic library constructed from tidal-flat sediment. Expression of the cloned biosynthetic genes in Escherichia coli led to the production of vibrioferrin, a siderophore originally reported for the marine bacterium Vibrio parahaemolyticus. To the best of our knowledge, this is the first example of heterologous production of a siderophore by biosynthetic genes cloned from a metagenomic library. The cloned cluster was one of the largest of the clusters obtained by functional screening. In this study, we demonstrated and extended the possibility of function-based metagenomic research.
通过对从潮滩沉积物构建的宏基因组文库进行功能筛选,克隆了一个由五个开放阅读框(ORF)组成的铁载体生物合成基因簇。将克隆的生物合成基因在大肠杆菌中表达,产生了弧菌素,这是一种最初报道来自海洋细菌副溶血性弧菌的铁载体。据我们所知,这是通过从宏基因组文库中克隆的生物合成基因异源生产铁载体的首个实例。克隆的基因簇是通过功能筛选获得的最大基因簇之一。在本研究中,我们证明并拓展了基于功能的宏基因组研究的可能性。