PreprintArticleVersion 1Preserved in Portico This version is not peer-reviewed
A High-throughput Method for Fingerprinting of Rhizobial Free Fatty Acids by the Combination of a Thin Film Chemical Deposition Technique and Matrix-assisted Laser Desorption/ionization Time-of Flight Mass Spectrometry
Version 1
: Received: 13 January 2020 / Approved: 13 January 2020 / Online: 13 January 2020 (13:25:49 CET)
How to cite:
Gladchuk, A.; Schumilina, J.; Kusnetsova, A.; Bureiko, K.; Tsarev, A.; Alexandrova, I.; Leonova, L.; Zhukov, V.A.; Tikhonovich, I.A.; Birkemeyer, C.; Podolskaya, E.; Frolov, A. A High-throughput Method for Fingerprinting of Rhizobial Free Fatty Acids by the Combination of a Thin Film Chemical Deposition Technique and Matrix-assisted Laser Desorption/ionization Time-of Flight Mass Spectrometry. Preprints2020, 2020010139. https://doi.org/10.20944/preprints202001.0139.v1
Gladchuk, A.; Schumilina, J.; Kusnetsova, A.; Bureiko, K.; Tsarev, A.; Alexandrova, I.; Leonova, L.; Zhukov, V.A.; Tikhonovich, I.A.; Birkemeyer, C.; Podolskaya, E.; Frolov, A. A High-throughput Method for Fingerprinting of Rhizobial Free Fatty Acids by the Combination of a Thin Film Chemical Deposition Technique and Matrix-assisted Laser Desorption/ionization Time-of Flight Mass Spectrometry. Preprints 2020, 2020010139. https://doi.org/10.20944/preprints202001.0139.v1
Gladchuk, A.; Schumilina, J.; Kusnetsova, A.; Bureiko, K.; Tsarev, A.; Alexandrova, I.; Leonova, L.; Zhukov, V.A.; Tikhonovich, I.A.; Birkemeyer, C.; Podolskaya, E.; Frolov, A. A High-throughput Method for Fingerprinting of Rhizobial Free Fatty Acids by the Combination of a Thin Film Chemical Deposition Technique and Matrix-assisted Laser Desorption/ionization Time-of Flight Mass Spectrometry. Preprints2020, 2020010139. https://doi.org/10.20944/preprints202001.0139.v1
APA Style
Gladchuk, A., Schumilina, J., Kusnetsova, A., Bureiko, K., Tsarev, A., Alexandrova, I., Leonova, L., Zhukov, V.A., Tikhonovich, I.A., Birkemeyer, C., Podolskaya, E., & Frolov, A. (2020). A High-throughput Method for Fingerprinting of Rhizobial Free Fatty Acids by the Combination of a Thin Film Chemical Deposition Technique and Matrix-assisted Laser Desorption/ionization Time-of Flight Mass Spectrometry. Preprints. https://doi.org/10.20944/preprints202001.0139.v1
Chicago/Turabian Style
Gladchuk, A., Ekaterina Podolskaya and Andrej Frolov. 2020 "A High-throughput Method for Fingerprinting of Rhizobial Free Fatty Acids by the Combination of a Thin Film Chemical Deposition Technique and Matrix-assisted Laser Desorption/ionization Time-of Flight Mass Spectrometry" Preprints. https://doi.org/10.20944/preprints202001.0139.v1
Abstract
Fatty acids (FAs) represent an important class of metabolites, impacting on membrane building blocks and cellular regulatory networks. In nature, prokaryotes are characterized with the most impressing FA structural diversity and the highest relative contents of free fatty acids (FFAs). Thereby, nitrogen-fixing bacteria (order Rhizobiales), often found in symbiosis with legumes, attract a special interest. Indeed, FAs impact on the structure of rhizobial nodulation factors, required for successful infection of plant root. Although the FA patterns can be addressed by GC- and LC-MS, these methods are time-consuming and suffer from compromised sensitivity, low stability of derivatives and artifacts. In contrast, MALDI-TOF-MS represents an excellent platform for high-efficient metabolite fingerprinting, also applicable to FFAs. Therefore, here we propose a simple and straightforward protocol for high-throughput relative quantification of FFAs in rhizobia by the combination of Langmuir technology and MALDI-TOF-MS, which is featured with high sensitivity, accuracy and precision of quantification. Here we propose a step by step procedure comprising rhizobia culturing, pre-cleaning, extraction, sample preparation, mass spectrometric analysis, data processing and post-processing. To demonstrate the analytical potential of the protocol we illustrate it by a case study – comparison the FFA metabolomes of two rhizobia species - Rhizobium leguminosarum and Sinorhizobium meliloti.
bacteria; barium monocarboxylates; chemical deposition techniques; free fatty acids (FFAs); Langmuir film technology; matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS); metabolic fingerprinting; rhizobia
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
,
,
,
,
*
