Purification of Arachidonic Acid from Mortierella Single-Cell Oil by Selective Esterification with Burkholderia cepacia Lipase

摘要

Purification of arachidonic acid(AA)from Mortierella alpina single-cell oil was attempted.The process comprised three steps:(I)preparation of FFA by nonselective hydrolysis of the oil with Alcaligenes sp.lipase;(II)elimination of long-chain saturated FA from the resulting FFA by urea adduct fractionation;and(iii)enrichment of AA through lipase-catalyzed selective esterification with lauryl alcohol(LauOH).In the third step,screening of industrially available lipases indicated that Burkholderia cepacia lipase(Lipase-PS,Amano Enzyme Inc.,Aichi,Japan)acted on AA more weakly than on other FA and was the most effective for enrichment of AA in the FFA fraction.When the FFA obtained by urea adduct fractionation were esterified with 2 molar equivalents of LauOH at 30 deg C for 16 h in a mixture with 20% water and 20 units(U)/g-mixture of Lipase-PS,the esterification reached 39% and the content of AA in the FFA fraction was raised from 61 to 86 wt%.To further increase the content of AA,unesterified FFA were allowed to react again under the same conditions as those in the first selective esterification except for the use of 50 U/g Lipase-PS.A series of procedures raised the content of AA to 97 wt% with a 49% recovery based on the initial content in the single-cell oil.These results indicated that the three-step process for selective esterification with Lipase-PS was effective for purifying AA from the single-ce[l oil.