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Selective enrichment of n−3 polyunsaturated fatty acids with C18–C20 acyl chain length from sardine oil using Pseudomonas fluorescens MTCC 2421 lipase

机译:使用荧光假单胞菌mTCC 2421脂肪酶从沙丁鱼油中选择性富集具有C18-C20酰基链长的n-3多不饱和脂肪酸

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摘要

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 6 h 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 6 h 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%.
机译:从荧光假单胞菌MTCC 2421纯化的细胞外脂肪酶用于在6小时后用二十碳五烯酸(20:5 n-3)和亚麻酸(18:3 n-3)分别使沙丁鱼油甘油三酸酯富集至35.28%和8.25%。水解。相应的n-6脂肪酸(18:2 n-6和20:4 n-6)显示出减少(分别为54.93%和50%)。结构与生物活性之间的关系分析表明,较低的疏水常数(log P值)为18:3 n-3和20:5 n-3(分别为5.65和5.85),导致它们对脂肪酶的耐水解性更高,从而使其富集脂肪酶催化水解后的甘油三酸酯馏分脂肪酶催化的沙丁鱼油水解6小时,然后在4°C下用甲醇将尿素分馏,分别得到分别含有42.50%20:5 n-3和10.31%18:3 n-3的游离脂肪酸。采用正己烷/乙酸乙酯(2:1,v / v)进行的中和氧化铝柱色谱纯化,得到20:5 n-3的高纯度(83.62%),而发现18:3 n-3可用正己烷/二氯甲烷(4:1,v / v)作为洗脱溶剂,最终纯度为75.31%。

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