Enzymatic analysis of positional distribution of fatty acids in solid fat by 1,3-selective transesterification with Candida antarctica lipase B
Enzymatic analysis of positional distribution of fatty acids in solid fat by 1,3-selective transesterification with Candida antarctica lipase B
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使用南极假丝酵母脂肪酶 B 进行 1,3-选择性酯交换反应，对固体脂肪中脂肪酸的位置分布进行酶法分析
DOI：
10.1007/s11746-014-2481-7
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Aki T.
中科院分区：
文献类型：
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作者：
Watanabe Y.;Sato S.;Sera S.;Sato C.;Yoshinaga K.;Nagai T.;Sato R.;Iwasaka H.;Aki T.
采用固定化南极念珠菌脂酶B （CALB）催化sn‐1(3)选择性醇解法，建立了固体三酰基甘油（TAG）中脂肪酸（FA）位置分布的分析方法。1份重量的固体脂肪和10份重量的乙醇（99.5%）加热使脂肪液化。加入重量为0.44份的CALB后，在50℃下振荡10 min，然后在30℃下振荡2.8 h。经过3 h的酯交换反应，2 - MAG的回收率为1。最大理论产率（33摩尔%）的85%，其中15%的损失是由于α - 2 - tosn - 1的酰基迁移(3)。回收率类似于在30°C下进行3小时的无溶剂醇解结构脂，1,3‐双棕榈酰，2‐油酰甘油和1,3‐二棕榈酰，2‐棕榈酰甘油。相比之下，几乎没有观察到酰基从msn‐1(3)tosn‐2迁移。由于检测到的酰基迁移只在sn‐2 tosn‐1(3)的方向上，而不是相反，因此建议通过气相色谱分析从酶反应混合物中回收的2‐MAG馏分来确定TAG的sn‐2位置的FA组成，并通过使用TAG和sn‐2位置的FA组成作为输入的质量平衡来确定sn‐1(3)位置的FA组成。该方法成功地应用于棕榈油和乳木果油，以及富含二十二碳六烯酸（DHA）的单细胞油。第一次是KH105。
A protocol for the analysis of the positional distribution of fatty acids (FA) in solid triacylglycerols (TAG) was developed usingsn‐1(3) selective alcoholysis catalyzed by immobilizedCandida antarcticalipase B (CALB). One part by weight of solid fat and ten parts by weight of ethanol (99.5 %) were warmed to liquefy the fat. After adding 0.44 parts by weight of CALB, the mixture was shaken at 50 °C for 10 min then at 30 °C for 2.8 h. The recovery of 2‐MAG after the 3‐h transesterification reaction wasca. 85 % of the maximum theoretical yield (33 mol%), with the loss of 15 % attributable to the acyl migration fromsn‐2 tosn‐1(3). The recovery was similar to that of the solvent‐free alcoholysis of structured lipids, 1,3‐dipalmitoyl, 2‐oleoyl glycerol and 1,3‐dioleoyl, 2‐palmitoyl glycerol, conducted at 30 °C for 3 h. In contrast, the acyl migration fromsn‐1(3) tosn‐2 was hardly observed. Because the detected acyl migration was only in the direction ofsn‐2 tosn‐1(3), and not vice versa, it is proposed to determine the FA composition of thesn‐2 position of TAG by the gas chromatographic analysis of 2‐MAG fraction recovered from the enzymatic reaction mixture, and the FA composition ofsn‐1(3) position by a mass balance using the FA composition of TAG and of thesn‐2 position as inputs. The procedure was successfully applied to palm oil and shea butter, and docosahexaenoic acid (DHA)‐rich single cell oil fromAurantiochytriumsp. KH105 for the first time.