利用大豆油脱臭馏出物酶法合成植物甾醇酯
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摘要
从大豆油脱臭馏出物中分别分离得到植物甾醇和脂肪酸,对植物甾醇的分离提取条件进行优化,确定甲醇-丙酮混合溶剂的比例为1∶2(V/V),溶剂原料比为3∶1(V/W),洗涤粗甾醇的溶剂为正己烷,分离出的脂肪酸主要为油酸、亚油酸等不饱和脂肪酸。通过物理吸附法固定CRL脂肪酶,考察不同载体固定脂肪酶的担载率和酶活力,确定大孔树脂HP20为最佳固定载体,酶活力最高且担载率最大。分离得到的植物甾醇和脂肪酸在固定化酶的催化作用下酯化合成植物甾醇酯,酯化率为45.11%,为日后各领域的应用提供了新的思路。
Phytosterol and fatty acid were isolated from soybean oil deodorizer distillate respectively.The extracting conditions of phytosterol were optimized and the optimal conditions were as follows: the ratio of methanol to acetone was 1 ∶ 2( V/V); the ratio of solvent to sample was 3∶ 1( V/W); n-hexane was selected as detergent.The isolated fatty acid mainly included oleic acid and linoleic acid.CRL lipase was immobilized by physical adsorption and immobilization degree and enzyme activity were determined as selecting indexes.HP20 resin was determined as the most suitable carrier because of its good performance. The synthesis of phytosterol fatty acid ester was carried out by the obtained phytosterol and fatty acid under the catalysis of immobilized enzyme,which provides a new pathway for the practice of all kinds of areas.
Phytosterol and fatty acid were isolated from soybean oil deodorizer distillate respectively.The extracting conditions of phytosterol were optimized and the optimal conditions were as follows: the ratio of methanol to acetone was 1 ∶ 2( V/V); the ratio of solvent to sample was 3∶ 1( V/W); n-hexane was selected as detergent.The isolated fatty acid mainly included oleic acid and linoleic acid.CRL lipase was immobilized by physical adsorption and immobilization degree and enzyme activity were determined as selecting indexes.HP20 resin was determined as the most suitable carrier because of its good performance. The synthesis of phytosterol fatty acid ester was carried out by the obtained phytosterol and fatty acid under the catalysis of immobilized enzyme,which provides a new pathway for the practice of all kinds of areas.
引文
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[19]Villeneuve P,Turon F,Caro Y,et al.Lipase-catalyzed synthesis of canola phytosterolsoleate esters as cholesterol lowering agents[J].Enzyme&Microbial Technology,2005,37(1):150-155.
[20]W He,C Jia.Lipase-catalyzed synthesis of phytostanyl esters in non-aqueous media[J].Journal of Molecular Catalysis BEnzymatic,2010,7(6):60-65.
[2]左玉.植物甾醇研究与应用[J].粮食与油脂,2012,5(7):1-4.
[3]许新德,邵斌.植物甾醇酯——一种新型的健康食品[J].食品添加剂,2004,29(8):25-28.
[4]雷炳福.油脂脱臭馏出物的组成与市场前景[J].中国油脂,2002,1(3):75-78.
[5]徐水祥,周淡宜,徐飞鹤,等.甲鱼油中多不饱和脂肪酸的提取与纯化研究[J].食品科学,2004,25(11):118-119.
[6]沈彩英.乌贼油中多不饱和脂肪酸的提取[J].中国油脂,2008,33(2):76-79.
[7]辛嘉英,李树本.脂肪酶的固定化及其在有机酶促反应中稳定性研究[J].分子催化,1999,13(2):103-108.
[8]马玲芝,钱俊青.离子交换树脂固定化脂肪酶催化合成蔗糖乙酯[J].浙江工业大学学报,2012,40(1):21-24.
[9]姜德生,龙胜亚,肖海燕,等.磁性壳聚糖微球的制备及其用作漆酶固定化载体[J].应用化学,2005,22(10):1122-1126.
[10]张琛,张宽朝,章琛,等.海藻酸钠包埋法制备固定化菠萝蛋白酶[J].生物加工过程,2009(6):31-34.
[11]许青青,金文彬,苏宝根,等.植物甾醇酯的化学合成及其分离研究进展[J].中国粮油学报,2014,3(6):120-128.
[12]许文林,沙鸥,钱俊红,等.混合植物甾醇中豆甾醇和β-谷甾醇的高效液相色谱分析[J].分析测试学报,2003,22(6):98-100.
[13]许文林,王岩.β-谷甾醇与马来酸酐酯化产物的表征及高效液相色谱分析[J].扬州大学学报,2006,3(9):32-35.
[14]Santas J,Codony R,Rafecas M.Phytosterols:Beneficial effects[J].Natural Products,2013,113(6):3437-3464.
[15]陈舒.植物甾醇酯微波加热合成的研究[D].西安:西安理工大学,2007.
[16]刘振春,孙慧娟.脂肪酶催化共轭亚油酸植物甾醇酯合成工艺的优化[J].西北农林科技大学学报,2014,42(6):173-179.
[17]陈陶声,居乃琥,陈石根,等.固定化酶理论与应用[M].北京:轻工业出版社1987:98-102.
[18]于殿宇,王腾宇,陈昊,等.生物酶法合成植物甾醇酯的研究[J].食品工业科技,2011,32(6):318-393.
[19]Villeneuve P,Turon F,Caro Y,et al.Lipase-catalyzed synthesis of canola phytosterolsoleate esters as cholesterol lowering agents[J].Enzyme&Microbial Technology,2005,37(1):150-155.
[20]W He,C Jia.Lipase-catalyzed synthesis of phytostanyl esters in non-aqueous media[J].Journal of Molecular Catalysis BEnzymatic,2010,7(6):60-65.