Selective enrichment of n-3 polyunsaturated fatty acids with C₁₈-C₂₀ acyl chain length from sardine oil using Pseudomonas fluorescens MTCC 2421 lipase
2009
Chakraborty, Kajal | Paul Raj, R.
An extracellular lipase purified from Pseudomonas fluorescens MTCC 2421 was used to enrich sardine oil triglycerides with eicosapentaenoic acid (20:5 n-3) and linolenic acid (18:3 n-3) to 35.28% and 8.25%, respectively, after 6h of hydrolysis. The corresponding n-6 fatty acids (18:2 n-6 and 20:4 n-6) exhibit a reduction (54.93% and 50%, respectively). Structure-bioactivity relationship analyses revealed that the lower hydrophobic (log P values) constants of 18:3 n-3 and 20:5 n-3 (5.65 and 5.85, respectively) result in their higher hydrolytic resistance towards lipase, leading to their enrichment in the triglyceride fraction after lipase-catalysed hydrolysis. Lipase-catalysed hydrolysis of sardine oil for 6h followed by urea fractionation at 4°C with methanol provided free fatty acids containing 42.50% 20:5 n-3 and 10.31% 18:3 n-3, respectively. Argentation neutral alumina column chromatography, using n-hexane/ethylacetate (2:1, v/v) resulted in 20:5 n-3 of high purity (83.62%), while 18:3 n-3 was found to be eluted with n-hexane/dichloromethane (4:1, v/v) as eluting solvent with a final purity of 75.31%.
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