Reversed-phase HPLC/electrospray ionization mass spectromety of phosphatidylglycerod molecular species in Pseudomonas fluorescens
2005
Taoka, Y.(Hokkaido Univ., Sapporo (Japan)) | Itabashi, Y.
Regio-specific analysis of molecular species of bacterial phosphatidylglycerols (PG; a mixture of 1,2-diacyl-sn-glycero-3-phospho-1'-sn-glycerol and 1,2-diacyl-sn-glycero-3-phospho-3'-sn-glycerol) was carried out using reversed-phase high-performance liquid chromatography (HPLC) in conjunction with electrospray ionization mass spectrometry (ESI-MS). For this purpose, the Gram negative bacterium Pseudomonas fluorescens IAMl2001 strain was used. The PG fraction was isolated from total lipids of P. fuorescens in the stationary phase and resolved into molecular species on an ODS column (25 cm x 4.6 mm i.d., 5 mum particle size) using methanol/ water/aqueous ammonium hydroxide (94: 6:0.1, v/v/v) as the mobile phase. On-line ESI-MS gave weak productions [M-RCO]-, [M-H-RCOOH]- and [RCOO]-, in addition to a prominent deprotonated [M-H]- molecule. The three productions derived from a fatty acid at the sn-2 position] showed greater intensities than those from the sn-1 position, by which individual molecular species could be identified The major molecular species (sn-1/sn-2) were 16:0/cis11, 12-methylenehexadecanoic acid (cyl7:0), 18:l/cyl7:0 and 16:0/18:l, of which the sum occupied about 73% of totalPG.
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