Identification of Unanticipated and Novel N-Acyl L-Homoserine Lactones (AHLs) Using a Sensitive Non-Targeted LC-MS/MS Method.
Identification of Unanticipated and Novel N-Acyl L-Homoserine Lactones (AHLs) Using a Sensitive Non-Targeted LC-MS/MS Method.
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DOI：
10.1371/journal.pone.0163469
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Amador-Noguez D
中科院分区：
文献类型：
--
作者：
Patel NM;Moore JD;Blackwell HE;Amador-Noguez D
n -酰基l -高丝氨酸内酯（ahl）构成了革兰氏阴性菌使用的群体感应信号分子的主要类别。在这里，我们报告了一种基于平行反应监测（PRM）的灵敏非靶向HPLC-MS/MS方法，用于鉴定和定量微生物样品中已知的、未预料到的和新的ahl。使用混合四极杆-高分辨率质谱分析仪，该方法集成了MS扫描和全离子碎片MS/MS扫描，可以同时检测AHL亲本离子质量，并在单次色谱运行中以高分辨率和高质量精度生成全质谱。我们应用该方法筛选了多种革兰氏阴性菌（即洋葱芽孢杆菌、延迟芽孢杆菌、胡萝卜芽孢杆菌、除草剂芽孢杆菌、stewartii芽孢杆菌、铜绿芽孢杆菌、金黄色芽孢杆菌和球形芽孢杆菌）产生AHL的情况，发现几乎所有革兰氏阴性菌产生的AHL数量都比之前报道的要多。此外，我们还在E. carotovora和P. stewartii中发现了一种罕见的AHL（即3-oxo-C7-HL），这种AHL的产生以前只在Serratia属和Yersinia属中观察到。最后，我们用我们的方法定量了洋葱芽孢杆菌、胡萝卜芽孢杆菌、除草剂芽孢杆菌、stewartii芽孢杆菌、铜绿芽孢杆菌、金黄色芽孢杆菌、非AHL产生菌大肠杆菌和革兰氏阳性杆菌枯草芽孢杆菌对AHL的降解。我们发现这些微生物对AHL的降解能力差异很大，其中枯草芽孢杆菌和胡萝卜芽孢杆菌是最好的降解者，并且观察到含有长酰基链（≥10个碳）的AHL的降解速度普遍高于含有短酰基链（≤6个碳）的AHL。
N-acyl L-homoserine lactones (AHLs) constitute a predominant class of quorum-sensing signaling molecules used by Gram-negative bacteria. Here, we report a sensitive and non-targeted HPLC-MS/MS method based on parallel reaction monitoring (PRM) to identify and quantitate known, unanticipated, and novel AHLs in microbial samples. Using a hybrid quadrupole-high resolution mass analyzer, this method integrates MS scans and all-ion fragmentation MS/MS scans to allow simultaneous detection of AHL parent-ion masses and generation of full mass spectra at high resolution and high mass accuracy in a single chromatographic run. We applied this method to screen for AHL production in a variety of Gram-negative bacteria (i.e. B. cepacia, E. tarda, E. carotovora, E. herbicola, P. stewartii, P. aeruginosa, P. aureofaciens, and R. sphaeroides) and discovered that nearly all of them produce a larger set of AHLs than previously reported. Furthermore, we identified production of an uncommon AHL (i.e. 3-oxo-C7-HL) in E. carotovora and P. stewartii, whose production has only been previously observed within the genera Serratia and Yersinia. Finally, we used our method to quantitate AHL degradation in B. cepacia, E. carotovora, E. herbicola, P. stewartii, P. aeruginosa, P. aureofaciens, the non-AHL producer E. coli, and the Gram-positive bacterium B. subtilis. We found that AHL degradation ability varies widely across these microbes, of which B. subtilis and E. carotovora are the best degraders, and observed that there is a general trend for AHLs containing long acyl chains (≥10 carbons) to be degraded at faster rates than AHLs with short acyl chains (≤6 carbons).
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影响因子：
--
作者：
Clasquin, Michelle F;Melamud, Eugene;Rabinowitz, Joshua D
通讯作者：
Rabinowitz, Joshua D
影响因子：
4.4
作者：
Burton, EO;Read, HW;Hickey, WJ
通讯作者：
Hickey, WJ
影响因子：
4.4
作者：
Huang, JJ;Han, JI;Leadbetter, JR
通讯作者：
Leadbetter, JR
DOI：
10.1073/pnas.1114125108
发表时间：
2011-10-04
影响因子：
11.1
作者：
Lindemann, Andrea;Pessi, Gabriella;Harwood, Caroline S.
通讯作者：
Harwood, Caroline S.
影响因子：
4.1
作者：
Fetzner, Susanne
通讯作者：
Fetzner, Susanne