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Isolation and Chemical Characterization of Lipid A from Gram-negative Bacteria
Published on: September 16, 2013
A novel procedure for the isolation of glycolipids from Bifidobacterium adolescentis 94 BIM using supercritical
Galina Novik1, Andrzej Gamian, José da Cruz Francisco
1Institute of Microbiology, National Academy of Sciences of Belarus, Kuprevich 2, 220141 Minsk, Belarus, Poland.
Abstract:
This study describes a novel isolation procedure for major glycolipids from Bifidobacterium adolescentis 94 BIM. The procedure consists of the use of supercritical carbon dioxide (scCO(2)) with hydro-methanolic solution as co-solvent. The major glycolipids were isolated using the following operating conditions: pressure, 30 MPa, co-solvent concentration, 10% (9:1, methanol/water, v/v), CO(2) flow rate, 5 g/min, extraction time and temperature, 2h and 55 degrees C, respectively. The reference glycolipids sample was prepared by classical organic solvent extraction followed by chromatographic purification. All isolates were characterized by TLC and the major glycolipids additionally by enzyme linked immunosorbent (ELISA). Sixty milligrams of glycolipids with similar immunoreactivity as the reference glycolipids were isolated from 1g of freeze-dried biomass (6% of yield).
