利用月桂酸和山茶油的酸解反应分析脂肪酶的位置专一性.docx

利用月桂酸和山茶油的酸解反应分析脂肪酶的位置专一性曹茜1韦伟1 张希2 冯凤琴1（浙江大学食品科学与营养系浙江省农产品加工技术研究重点实验室1，杭州 310058）（云南中医学院2，昆明650500）摘要脂肪酶的位置专一性在结构酯的合成和油脂改性中具有重要意义。由于水体系和非水体系的差异，传统的水解判定法得出的专一性与该酶在合成反应中的表现可能并不一致。本研究利用月桂酸和山茶油的酸解反应来达到直接评估酶在无溶剂体系中位置专一性的目的。反应产物的脂肪酸组成和位置分布由气相色谱测定。通过此方法，Lipozyme RM IM、L02、L03和L04被鉴定为sn-1(3)位专一性，L01为弱专一性，Novozym 435近似无专一性。通过替换酶的底物，模型反应的可预测性得到验证。根据酸解法和水解法结果的对比分析，两种条件下酶的位置专一性通常是相同的，除了易受到溶剂体系影响的Novozym 435。因此，新方法能够避免水解判定结果在合成反应中应用的局限性。此外，它还降低了对底物纯度的要求，这意味着其能在低成本下用于脂肪酶的大规模筛选。关键词脂肪酶位置专一性山茶油酸解合成中图分类号：TQ925+.6 文献标识码：A 文章编号：1003-0174（）- - Analysis of regioselectivity of lipases using acidolysis of lauric acid and camellia oilCao Xi1Wei Wei1 Zhang Xi2Feng Fengqin1(Department of Food Science and Nutrition, Zhejiang UniversityZhejiang Key Laboratory for Agro-Food Processing1, Hangzhou 310058)(Yunnan College of Traditional Chinese Medicine2, Kunming 650500)AbstractRegioselectivity of lipases plays a crucial role in structured lipids synthesis and oil modification. Because of differences between aqueous and non-aqueous conditions, regioselectivity analyzed by the conventional hydrolysis method might not always match that in synthesis reactions. In this paper, acidolysis of lauric acid and camellia oil was developed to directly evaluate regioselectivity in solvent-free media. Fatty acid composition and positional distribution of reaction products were detected by gas chromatography. Through this protocol, Lipozyme RM IM, L02, L03 and L04 were identified as sn-1(3) regioselective, L01 was partiallyselective, and Novozym 435 was nearly non-selective. Predictability of the model reaction has been verified by replacing substrates of lipases. According to the comparative analysis between the acidolysis method and traditional hydrolysis one, regioselectivity in two situationsis generally the same, except selectivity of Novozym 435 which is susceptible to solvent systems. Therefore the present method avoids limits of hydrolysis evaluation systems in synthesis reactions. And moreover i