A proteomic survey of nonribosomal peptide and polyketide biosynthesis in actinobacteria.
A proteomic survey of nonribosomal peptide and polyketide biosynthesis in actinobacteria.
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DOI：
10.1021/pr2009115
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发表时间：
2012-01-01
影响因子：
4.4
通讯作者：
Kelleher, Neil L.
中科院分区：
文献类型：
--
作者：
Chen, Yunqiu;Ntai, Ioanna;Ju, Kou-San;Unger, Michelle;Zamdborg, Leonid;Robinson, Sarah J.;Doroghazi, James R.;Labeda, David P.;Metcalf, William W.;Kelleher, Neil L.
关键词：
放线菌如链霉菌以其产生生物活性天然产物的能力而闻名，所述生物活性天然产物包括非核糖体肽（NRP）和聚酮化合物（PK）。基因组测序的出现揭示了更大的次级代谢的遗传库，这些基因簇的大多数小分子产物仍然未知。在这里，我们采用了一种“蛋白质优先”的方法，称为PrISM（蛋白质组学研究的次级代谢），筛选26个未测序的放线菌，使用基于质谱的蛋白质组学表达的非核糖体肽合成酶或聚酮酶的靶向检测。对原始PrISM筛选方法的改进（Nature Biotechnology，2009，27，951 - 956），例如改进的从头肽测序，已经使得能够从六种菌株发现十个NRPS/PKS基因簇。利用生物合成酶和它们产生的次级代谢产物的协同作用，将两种天然产物与它们以前的“孤儿”基因簇相关联。这项工作已经证明了基于蛋白质组学的策略用于筛选放线菌（通常>8 Mbp，高GC基因组）与先前使用的杆菌（2-4 Mbp基因组）中的NRP/PK产生的可行性。
Actinobacteria such as streptomycetes are renowned for their ability to produce bioactive natural products including nonribosomal peptides (NRPs) and polyketides (PKs). The advent of genome sequencing has revealed an even larger genetic repertoire for secondary metabolism with most of the small molecule products of these gene clusters still unknown. Here, we employed a “protein-first” method called PrISM (Proteomic Investigation of Secondary Metabolism) to screen 26 unsequenced actinomycetes using mass spectrometry-based proteomics for the targeted detection of expressed nonribosomal peptide synthetases or polyketide synthases. Improvements to the original PrISM screening approach (Nature Biotechnology, 2009, 27, 951 – 956), e.g. improved de novo peptide sequencing, have enabled the discovery of ten NRPS/PKS gene clusters from six strains. Taking advantage of the concurrence of biosynthetic enzymes and the secondary metabolites they generate, two natural products were associated with their previously ‘orphan’ gene clusters. This work has demonstrated the feasibility of a proteomics-based strategy for use in screening for NRP/PK production in actinomycetes (often >8 Mbp, high GC genomes) versus the bacilli (2–4 Mbp genomes) used previously.
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影响因子：
7
作者：
Li, Ruiqiang;Zhu, Hongmei;Wang, Jun
通讯作者：
Wang, Jun
影响因子：
11.9
作者：
Nett M;Ikeda H;Moore BS
通讯作者：
Moore BS
DOI：
10.1099/00221287-43-2-159
发表时间：
1966-01-01
期刊：
JOURNAL OF GENERAL MICROBIOLOGY
影响因子：
--
作者：
STANIER, RY;PALLERON.NJ;DOUDOROF.M
通讯作者：
DOUDOROF.M
影响因子：
46.9
作者：
Meng, FY;Cargile, BJ;Kelleher, NL
通讯作者：
Kelleher, NL
影响因子：
7
作者：
Chaisson, Mark J.;Brinza, Dumitru;Pevzner, Pavel A.
通讯作者：
Pevzner, Pavel A.