Guidelines for metabolomics-guided transposon mutagenesis for microbial natural product discovery.
Guidelines for metabolomics-guided transposon mutagenesis for microbial natural product discovery.
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DOI：
10.1016/bs.mie.2021.11.020
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发表时间：
2022
影响因子：
--
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--
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培养微生物的自然产物产量与其生物信息预测的生物合成潜力之间存在着巨大的差异，因此微生物储藏库中包含的大部分分子多样性尚未被发现。其中一个主要原因是天然产物生物合成基因簇(BGC)在标准实验室条件下表达不足。已经开发了几种方法来增加这种“神秘的”BGC的产量。其中，我们最近实现了质谱学引导的转座子突变，这是一种正向遗传筛选，通过质谱学读出的显示隐蔽代谢物的刺激生物合成的突变体从转座子突变体库中选择。在这里，我们以伯克霍尔德氏菌为例，为建立转座子突变体文库，通过质谱学测量代谢组清单，执行比较代谢组学以优先从突变体库中筛选神秘的天然产物，以及分离和鉴定通过诱变产生的新的天然产物提供指南。这种方法的应用将有助于从神秘的BGC中获得新的天然产物，并识别参与其全球调控的基因。
There is a great discrepancy between the natural product output of cultured microorganisms and their bioinformatically predicted biosynthetic potential, such that most of the molecular diversity contained within microbial reservoirs has yet to be discovered. One of the primary reasons is insufficient expression of natural product biosynthetic gene clusters (BGCs) under standard laboratory conditions. Several methods have been developed to increase production from such “cryptic” BGCs. Among these, we recently implemented mass spectrometry-guided transposon mutagenesis, a forward genetic screen in which mutants that exhibit stimulated biosynthesis of cryptic metabolites, as read out by mass spectrometry, are selected from a transposon mutant library. Herein, we use Burkholderia gladioli as an example and provide guidelines for generating transposon mutant libraries, measuring metabolomic inventories through mass spectrometry, performing comparative metabolomics to prioritize cryptic natural products from the mutant library, and isolating and characterizing novel natural products elicited through mutagenesis. Application of this approach will be useful in both accessing novel natural products from cryptic BGCs and identifying genes involved in their global regulation.
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影响因子：
3.4
作者：
Bachmann, Brian O.;Van Lanen, Steven G.;Baltz, Richard H.
通讯作者：
Baltz, Richard H.
DOI：
10.1073/pnas.1714381115
发表时间：
2017-12-26
影响因子：
11.1
作者：
Amos, Gregory C. A.;Awakawa, Takayoshi;Jensen, Paul R.
通讯作者：
Jensen, Paul R.
影响因子：
3.2
作者：
Mao, Dainan;Yoshimura, Aya;Seyedsayamdost, Mohammad R.
通讯作者：
Seyedsayamdost, Mohammad R.
影响因子：
16.6
作者：
通讯作者：
--
影响因子：
8.4
作者：
Guo, Fang;Xiang, Sihai;Yang, Kegian
通讯作者：
Yang, Kegian