Monitoring of monooctanoylphosphatidylcholine synthesis by enzymatic acidolysis between soybean phosphatidylcholine and caprylic acid by thin-layer chromatography with a flame ionization detector.
Thin-layer chromatography with a flame ionization detector (TLC-FID) was used for monitoring the production of structured phospholipids (ML type: L, long-chain fatty acids; M, medium-chain fatty acids) by enzyme-catalyzed acidolysis between soybean phosphatidylcholine (PC) and caprylic acid. It was found that the structured PC fractionated into two to three distinct bands on both plate thin-layer chromatography (TLC) and Chromarod TLC. These three bands represented PC of the LL type, ML type, and MM type, respectively. The TLC-FID method was applied in the present study to examine the influence of enzyme dosage, reaction temperature, solvent amount, reaction time, and substrate ratio (caprylic acid/PC, mol/mol) on formation of ML-type PC in a batch reactor with Thermomyces lanuginosa lipase as the catalyst. The formation of ML-type PC was dependent on all parameters examined except for the substrate ratio. The ML-type PC content increased with increasing enzyme dosage, reaction temperature, solvent amount, and reaction time. The substrate ratio had no significant effect on the formation of ML-type PC within the tested range (3-15 mol/mol). The formation of MM-type PC was observed in some experiments, indicating that acyl migration is taking place during reaction since the lipase is claimed to be 1,3-specific. The TLC-FID method offers a simple and cheap technique for elucidation of product and byproduct formation during enzyme-catalyzed reactions for production of phospholipids containing mixtures of long- and medium-chain fatty acids.
Vikbjerg
Anders F
AF
BioCentrum-DTU, Technical University of Denmark, DK 2800-Lyngby, Denmark. afv@biocentrum.dtu.dk
Mu
Huiling
H
Xu
Xuebing
X
D002210:Caprylates / Q000378:metabolism*; D002855:Chromatography, Thin Layer / Q000379:methods*; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism*; D010743:Phospholipids / Q000096:biosynthesis* / Q000737:chemistry; D013025:Glycine max / Q000737:chemistry*
D002210:Caprylates; D005227:Fatty Acids; D010713:Phosphatidylcholines; D010743:Phospholipids; D008049:Lipase; C031492:octanoic acid