Untargeted Identification of Alkyne-Containing Natural Products Using Ruthenium-Catalyzed Azide Alkyne Cycloaddition Reactions Coupled to LC-MS/MS.
Untargeted Identification of Alkyne-Containing Natural Products Using Ruthenium-Catalyzed Azide Alkyne Cycloaddition Reactions Coupled to LC-MS/MS.
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DOI：
10.1021/acs.jnatprod.1c00798
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发表时间：
2022-01-28
影响因子：
5.1
通讯作者：
Philmus B
中科院分区：
文献类型：
--
作者：
Back D;Shaffer BT;Loper JE;Philmus B
含有炔的天然产物已经从植物、昆虫、藻类、真菌和细菌中鉴定出来。这类天然产物具有多种生物活性。多炔是含有共轭炔的炔属天然产物的子类，并且由于它们在纯化过程中分解的事实而在天然产物数据库中代表性不足。在这里，我们报告了一个工作流程，利用炔叠氮化物环加成（AAC）反应，然后进行LC-MS/MS分析，以确定炔属天然产物。在这份报告中，我们证明了炔叠氮化物与对溴苄基叠氮化物的环加成反应导致对溴苄基取代的三唑产物，其片段为一个共同的溴化tropyridine离子。在优化MS/MS条件后，我们能够鉴定以10 μg/ml浓度掺入鱼腥藻PCC 7120提取物中的合成炔。然后，我们成功地确定了已知的天然产物fischerellin A的提取物Fischerella muscicola PCC 9339。最后，我们通过基因组挖掘和RuAAC反应的组合鉴定了最近鉴定的来自假单胞菌Protegens Pf-5的天然产物Protegenin A和C。这是首次报道RuAAC反应用于检测炔类天然产物。我们还比较了CuAAC和RuAAC反应，发现CuAAC反应产生的副产物比RuAAC少，但仅限于含末端炔的化合物。相比之下，RuAAC能够识别末端和内部炔属天然产物，但需要通过创建排除列表从分析中排除副产物。我们认为，CuAAC和RuAAC反应耦合LC-MS/MS代表了一种非靶向鉴定炔类天然产物的方法，但每种方法都有优点和缺点。
Alkyne containing natural products have been identified from plants, insects, algae, fungi, and bacteria. This class of natural products have been characterized as having a variety of biological activities. Polyynes are a sub-class of acetylenic natural products that contain conjugated alkynes and are underrepresented in natural product databases due to the fact that they decompose during purification. Here we report a workflow that utilizes alkyne azide cycloaddition (AAC) reactions followed by LC-MS/MS analysis to identify acetylenic natural products. In this report, we demonstrate that alkyne azide cycloaddition reactions with p-bromobenzyl azide result in p-bromobenzyl substituted triazole products that fragment to a common brominated tropylium ion. We were able to identify a synthetic alkyne spiked into the extract of Anabaena sp. PCC 7120 at a concentration of 10 μg/ml after optimization of MS/MS conditions. We then successfully identified the known natural product fischerellin A in the extract of Fischerella muscicola PCC 9339. Lastly, we identified the recently identified natural products protegenins A and C from Pseudomonas protegens Pf-5 through a combination of genome mining and RuAAC reactions. This is the first report of RuAAC reactions to detect acetylenic natural products. We also compare CuAAC and RuAAC reactions and find that CuAAC reactions produce less by-products compared to RuAAC but is limited to terminal alkyne containing compounds. In contrast, RuAAC is capable of identification of both terminal and internal acetylenic natural products, but by-products need to be eliminated from analysis by creation of an exclusion list. We believe that both CuAAC and RuAAC reactions coupled to LC-MS/MS represent a method for the untargeted identification of acetylenic natural products, but each method has strengths and weaknesses.
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影响因子：
5.1
作者：
Mejia EJ;Magranet LB;De Voogd NJ;Tendyke K;Qiu D;Shen YY;Zhou Z;Crews P
通讯作者：
Crews P
影响因子：
5.1
作者：
Brumley, David;Spencer, Kara A.;Luesch, Hendrik
通讯作者：
Luesch, Hendrik
DOI：
10.1038/nbt1183-784
发表时间：
1983-01-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
SIMON, R;PRIEFER, U;PUHLER, A
通讯作者：
PUHLER, A
影响因子：
15
作者：
Boren, Brant C.;Narayan, Sridhar;Fokin, Valery V.
通讯作者：
Fokin, Valery V.
影响因子：
3.2
作者：
Beresovsky, Dina;Hadas, Ora;Carmeli, Shmuel
通讯作者：
Carmeli, Shmuel