克隆和鉴定一个环境 DNA 来源的基因簇，该基因簇编码抗肿瘤物质 BE-54017 的生物合成。
Cloning and characterization of an environmental DNA-derived gene cluster that encodes the biosynthesis of the antitumor substance BE-54017.
机构信息
Laboratory of Genetically Encoded Small Molecules, The Rockefeller University, Howard Hughes Medical Institute, 1230 York Avenue, New York, New York 10065, USA.
出版信息
J Am Chem Soc. 2011 Jul 6;133(26):9996-9. doi: 10.1021/ja2022653. Epub 2011 May 4.
Abstract
Soil is predicted to contain thousands of unique bacterial species per gram. Soil DNA libraries represent large reservoirs of biosynthetic diversity from which diverse secondary metabolite gene clusters can be recovered and studied. The screening of an archived soil DNA library using primers designed to target oxytryptophan dimerization genes allowed us to identify and functionally characterize the first indolotryptoline biosynthetic gene cluster. The recovery and heterologous expression of an environmental DNA-derived gene cluster encoding the biosynthesis of the antitumor substance BE-54017 is reported here. Transposon mutagenesis identified two monooxygenases, AbeX1 and AbeX2, as being responsible for the transformation of an indolocarbazole precursor into the indolotryptoline core of BE-54017.
摘要
土壤预计每克含有数千种独特的细菌物种。土壤 DNA 文库代表了生物合成多样性的巨大资源库，可以从中回收和研究各种次级代谢物基因簇。使用针对氧色氨酸二聚化基因设计的引物筛选存档的土壤 DNA 文库，使我们能够鉴定和功能表征第一个吲哚洛特里品啉生物合成基因簇。本文报道了从环境 DNA 衍生的基因簇中恢复和异源表达编码抗肿瘤物质 BE-54017 的生物合成。转座子诱变鉴定出两个单加氧酶 AbeX1 和 AbeX2，它们负责将吲哚咔唑前体转化为 BE-54017 的吲哚洛特里品啉核心。
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