Research output: Contribution to journal › Article › peer-review
High-Throughput Fingerprinting of Rhizobial Free Fatty Acids by Chemical Thin-Film Deposition and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. / Gladchuk, Aleksey ; Shumilina, Julia ; Kusnetsova , Alena; Bureiko , Ksenia ; Billig, Susan; Tsarev , Alexander ; Alexandrova , Irina ; Leonova, Larisa ; Zhukov, Vladimir A. ; Tikhonovich , Igor A. ; Birkemeyer, Claudia ; Podolskaya, Ekaterina ; Frolov, Andrej .
In: Methods and Protocols, Vol. 3, No. 2, 36, 06.2020, p. 1-17.Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - High-Throughput Fingerprinting of Rhizobial Free Fatty Acids by Chemical Thin-Film Deposition and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry
AU - Gladchuk, Aleksey
AU - Shumilina, Julia
AU - Kusnetsova , Alena
AU - Bureiko , Ksenia
AU - Billig, Susan
AU - Tsarev , Alexander
AU - Alexandrova , Irina
AU - Leonova, Larisa
AU - Zhukov, Vladimir A.
AU - Tikhonovich , Igor A.
AU - Birkemeyer, Claudia
AU - Podolskaya, Ekaterina
AU - Frolov, Andrej
N1 - Publisher Copyright: © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2020/6
Y1 - 2020/6
N2 - Fatty acids (FAs) represent an important class of metabolites, impacting on membrane building blocks and signaling compounds in cellular regulatory networks. In nature, prokaryotes are characterized with the most impressing FA structural diversity and the highest relative content of free fatty acids (FFAs). In this context, nitrogen-fixing bacteria (order Rhizobiales), the symbionts of legumes, are particularly interesting. Indeed, the FA profiles influence the structure of rhizobial nodulation factors, required for successful infection of plant root. Although FA patterns can be assessed by gas chromatography—(GC-) and liquid chromatography—mass spectrometry (LC-MS), sample preparation for these methods is time-consuming and quantification suffers from compromised sensitivity, low stability of derivatives and artifacts. In contrast, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (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 combination of Langmuir technology and MALDI-TOF-MS featuring a high sensitivity, accuracy and precision of quantification. We describe a step-by-step procedure comprising rhizobia culturing, pre-cleaning, extraction, sample preparation, mass spectrometric analysis, data processing and post-processing. As a case study, a comparison of the FFA metabolomes of two rhizobia species—Rhizobium leguminosarum and Sinorhizobium meliloti, demonstrates the analytical potential of the protocol.
AB - Fatty acids (FAs) represent an important class of metabolites, impacting on membrane building blocks and signaling compounds in cellular regulatory networks. In nature, prokaryotes are characterized with the most impressing FA structural diversity and the highest relative content of free fatty acids (FFAs). In this context, nitrogen-fixing bacteria (order Rhizobiales), the symbionts of legumes, are particularly interesting. Indeed, the FA profiles influence the structure of rhizobial nodulation factors, required for successful infection of plant root. Although FA patterns can be assessed by gas chromatography—(GC-) and liquid chromatography—mass spectrometry (LC-MS), sample preparation for these methods is time-consuming and quantification suffers from compromised sensitivity, low stability of derivatives and artifacts. In contrast, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (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 combination of Langmuir technology and MALDI-TOF-MS featuring a high sensitivity, accuracy and precision of quantification. We describe a step-by-step procedure comprising rhizobia culturing, pre-cleaning, extraction, sample preparation, mass spectrometric analysis, data processing and post-processing. As a case study, a comparison of the FFA metabolomes of two rhizobia species—Rhizobium leguminosarum and Sinorhizobium meliloti, demonstrates the analytical potential of the protocol.
KW - Bacteria
KW - barium monocarboxylates
KW - chemical deposition technique
KW - free fatty acids (FFAs)
KW - Langmuir film technology
KW - matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS)
KW - metabolic fingerprinting
KW - rhizobia
KW - Barium monocarboxylates
KW - Rhizobia
KW - Free fatty acids (FFAs)
KW - Metabolic fingerprinting
KW - Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS)
KW - Chemical deposition technique
UR - https://www.mdpi.com/2409-9279/3/2/36
UR - http://www.scopus.com/inward/record.url?scp=85089848948&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/5ee4ab8f-f23b-35e0-b652-90156208fad0/
U2 - 10.3390/mps3020036
DO - 10.3390/mps3020036
M3 - Article
VL - 3
SP - 1
EP - 17
JO - Methods and Protocols — Open Access Journal
JF - Methods and Protocols — Open Access Journal
SN - 2409-9279
IS - 2
M1 - 36
ER -
ID: 53423664