酶催化法合成单硬脂酸甘油酯
摘要
单硬脂酸甘油酯是一种重要的多元醇型非离子表面活性剂。本研究以固定化1,3-特异性脂肪酶(Lipozyme~(?) IM)为催化剂，催化硬脂酸甲酯转酯化合成单硬脂酸甘油酯。研究表明，固定化1,3-特异性脂肪酶催化硬脂酸甲酯转酯化反应制单硬脂酸甘油酯合适的反应条件为：pH7.0、甘油含水量为8％、酶用量为硬脂酸甲酯的20％、甘油与硬脂酸甲酯的配比为1.5:1、反应温度为55℃、反应时间为34小时。在这一反应条件下，硬脂酸甲酯的转化率为92.3％，单甘酯收率为90.8％；酶通过分离回收后重复利用，酶的消耗量为硬脂酸甲酯量的1.0％。
对固定化1,3-特异性脂肪酶催化硬脂酸甲酯和甘油转酯化合成的粗产品进行了结晶分离，研究所得合适的结晶分离条件为：溶剂乙醇与粗单甘酯质量比为1.5:1；冷却结晶温度为20℃。经分离提纯后，所得最终产物中单硬脂酸甘油酯的含量为87.7％，其总收率为84.1％。
同时，对脂肪酶催化硬脂酸甲酯转酯化合成单硬脂酸甘油酯的反应机理及动力学进行了研究。研究表明，该酶催化反应符合双底物酶反应的乒乓反应机制，其动力学方程为：
其中，三个动力学参数v_m=0.154μmol／(g·s)，K_(mB)=0.593mmol／g，K_(mA)=0.367mmol／g。
研究表明，合成反应条件温和、产品纯度高、操作简便，为高纯度单硬脂酸甘油酯的合成提供了一个新的合成方法。
The Monoglyceride is a kind of important polyol non-ionic surface active agent. Highly pure monoglycerides was prepared by immobilized 1,3-specific lipase (Lipozyme IM) from methyl stearate in non-solvent system. It was proved that the proper conditions of synthesis reaction: pH was 7.0, the water was 8% based on glycerin, lipase was 20% based on methyl stearate, the ratio of glycerin to methyl stearate was 1.5, the reaction temperature was 55 D, and the reaction time was 34 hours. Under the conditions, the fractional conversion of methyl stearate was 92.3% and the monoglycerides yield was 90.8% in synthesize reaction. By reclamation, the amount of consumed immobilized lipase was 1.0% to methyl stearate .
The monoglycerides products were separated and purified by crystallization. The crystallization temperatures was 20 , the ratio of ethanol to the mixtures was 1.5. The yield of products was 84.1% and the content of monoglycerides was 87.7% in the final products.
The kinetics of lipase-catalyzed interesterification of glycerol and methyl stearate was also studied. It was proved that this reaction mechanism is Ping Pong between two reactants. The equation is as follows.
The vm , KmB and KmA are 0.154 umol/(g.s), 0.593 mmol/g and 0.367 mmol/g, respectively.
The investigation showed that the sum yield of monoglyceride under the studied conditions was 84.1%, and content of monoglyceride was 87.7% in final product. The reaction was processed under mild conditions, and produced high pure of monoglyceride product, and operation was easy. This study provided a new method of preparing high pure monoglycerides by enzymatic intransesterification of methyl stearate.
对固定化1,3-特异性脂肪酶催化硬脂酸甲酯和甘油转酯化合成的粗产品进行了结晶分离，研究所得合适的结晶分离条件为：溶剂乙醇与粗单甘酯质量比为1.5:1；冷却结晶温度为20℃。经分离提纯后，所得最终产物中单硬脂酸甘油酯的含量为87.7％，其总收率为84.1％。
同时，对脂肪酶催化硬脂酸甲酯转酯化合成单硬脂酸甘油酯的反应机理及动力学进行了研究。研究表明，该酶催化反应符合双底物酶反应的乒乓反应机制，其动力学方程为：
其中，三个动力学参数v_m=0.154μmol／(g·s)，K_(mB)=0.593mmol／g，K_(mA)=0.367mmol／g。
研究表明，合成反应条件温和、产品纯度高、操作简便，为高纯度单硬脂酸甘油酯的合成提供了一个新的合成方法。
The Monoglyceride is a kind of important polyol non-ionic surface active agent. Highly pure monoglycerides was prepared by immobilized 1,3-specific lipase (Lipozyme IM) from methyl stearate in non-solvent system. It was proved that the proper conditions of synthesis reaction: pH was 7.0, the water was 8% based on glycerin, lipase was 20% based on methyl stearate, the ratio of glycerin to methyl stearate was 1.5, the reaction temperature was 55 D, and the reaction time was 34 hours. Under the conditions, the fractional conversion of methyl stearate was 92.3% and the monoglycerides yield was 90.8% in synthesize reaction. By reclamation, the amount of consumed immobilized lipase was 1.0% to methyl stearate .
The monoglycerides products were separated and purified by crystallization. The crystallization temperatures was 20 , the ratio of ethanol to the mixtures was 1.5. The yield of products was 84.1% and the content of monoglycerides was 87.7% in the final products.
The kinetics of lipase-catalyzed interesterification of glycerol and methyl stearate was also studied. It was proved that this reaction mechanism is Ping Pong between two reactants. The equation is as follows.
The vm , KmB and KmA are 0.154 umol/(g.s), 0.593 mmol/g and 0.367 mmol/g, respectively.
The investigation showed that the sum yield of monoglyceride under the studied conditions was 84.1%, and content of monoglyceride was 87.7% in final product. The reaction was processed under mild conditions, and produced high pure of monoglyceride product, and operation was easy. This study provided a new method of preparing high pure monoglycerides by enzymatic intransesterification of methyl stearate.
引文
1．张万福．食品乳化剂．北京：中国轻工业出版社．1993．110-111．
2．陆用海，胡征宇．工业脂肪酸及其应用．北京：中国轻工业出版社．1992．406-408．
3. Sonntag N O V. Glycerolysis of fats and methyl esters—status, review and critique. J Am Oil Chem Soc. 1982 (59) A. 795-802.
4．时宏，郭洪．面向21世纪的分子蒸馏单硬脂酸甘油酯工业．中国油脂．2000．25(4)．38-43．
5．汪多仁．现代日用化工产品．北京：冶金工业出版社．2000．285．
6．张万福．食品乳化剂．北京：中国轻工业出版社．1993．117-136．
7．张万福．食品乳化剂．北京：中国轻工业出版社．1993．290-603．
8．汪多仁．现代日用化工产品．北京：冶金工业出版社．2000．288-290．
9．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．中国轻工业出版社．1991．308-309．
10．李健．分子蒸馏单硬脂酸甘油酯的应用．表面活性剂工业．1998(3)．40-41．
11．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．中国轻工业出版社．1991．303-305．
12．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．中国轻工业出版社．1991．305-306．
13．任春华．脂肪酸单硬脂酸甘油酯的合成与应用．泸天化科技．1993(3)．21-27．
14．张逸伟，卢世．由硬脂酸钠和环氧氯丙烷合成单硬脂酸甘油酯．化学试剂．1997．19(1)．48-50．
15．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．北京：中国轻工业出版社．1991．306．
16．石祖芸，吕德水等．高纯度单硬脂酸甘油酯合成研究．化学反应工程与工艺．1995．11(2)．208-211．
17．张树政．酶制剂工业(上)．北京：科学出版社．1984．5．
18．童海宝．酶催化——催化反应的新领域．精细与专用化学品．1993(13)．6．
19．沈寅初．生物技术在化学工业中的应用．上海化工．1998(12)．35-38．
20．雷武．令人瞩目的生物反应技术．化工之友．1999(4)．26-27．
21．周昕，杨丽等．生物酶在表面活性剂合成及改性中的应用．日用化学工业．1999(3)．60—61．
22．唐平，雕鸿荪．用于油脂改性的1，3-定向脂肪酶．中国油脂．1996．2l(4)．4-9．
23．谷利伟，赵金兰．酶技术在油脂工业中的应用．中国油脂．1998．23(5)．43-45．
24．吴可克，程菊花，高培焕．酶促法合成甘油单酸酯．表面活性剂工业．1996(4)．28-29．
25．黄显慈．选择性合成单双甘油酯．四川化工．1995(4)．52-54．
26．王明．油脂酶催化甘油解制备甘油单酯产率的进一步提高．日用化学工业译丛．1992(1)．19-22．
27．王万绪．油脂的高产率酶催化甘油解．日用化学工业译丛．1992(1)．15-18．
28．彭立风，谭天伟．脂肪酶催化棕榈油甘油解反应合成单硬脂酸甘油酯．北京化工大学学报．1998．25(2)．6-10．
29．彭立风，谭天伟．有机相脂肪酶催化合成单硬脂酸甘油酯的研究进展．化学世界．1998(3)．119-126．
30．罗贵民．有机溶剂中的酶催化反应．精细与专用化学品．1999(5)．12-13．
31．姚传义，俞耀庭等．反胶团酶反应研究进展．化工进展．1999(5)．22-25．
32．刘俊康，刘瑛等．微乳凝胶固定化酶催化长链脂肪酸酯的合成．无锡轻工大学学报．2000．19(4)．361-364．
33．吴姣莲．牛油的固相酶催化甘油解生产高产率甘油单酯．日用化学工业译丛．1991(6)．18-22．
34．阎建民，马润宇．乳膜技术及其在酶化工的应用．化工进展．1999(5)．35-38．
2．陆用海，胡征宇．工业脂肪酸及其应用．北京：中国轻工业出版社．1992．406-408．
3. Sonntag N O V. Glycerolysis of fats and methyl esters—status, review and critique. J Am Oil Chem Soc. 1982 (59) A. 795-802.
4．时宏，郭洪．面向21世纪的分子蒸馏单硬脂酸甘油酯工业．中国油脂．2000．25(4)．38-43．
5．汪多仁．现代日用化工产品．北京：冶金工业出版社．2000．285．
6．张万福．食品乳化剂．北京：中国轻工业出版社．1993．117-136．
7．张万福．食品乳化剂．北京：中国轻工业出版社．1993．290-603．
8．汪多仁．现代日用化工产品．北京：冶金工业出版社．2000．288-290．
9．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．中国轻工业出版社．1991．308-309．
10．李健．分子蒸馏单硬脂酸甘油酯的应用．表面活性剂工业．1998(3)．40-41．
11．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．中国轻工业出版社．1991．303-305．
12．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．中国轻工业出版社．1991．305-306．
13．任春华．脂肪酸单硬脂酸甘油酯的合成与应用．泸天化科技．1993(3)．21-27．
14．张逸伟，卢世．由硬脂酸钠和环氧氯丙烷合成单硬脂酸甘油酯．化学试剂．1997．19(1)．48-50．
15．王福海，陈海等．硬脂酸及脂肪酸衍生物生产工艺．北京：中国轻工业出版社．1991．306．
16．石祖芸，吕德水等．高纯度单硬脂酸甘油酯合成研究．化学反应工程与工艺．1995．11(2)．208-211．
17．张树政．酶制剂工业(上)．北京：科学出版社．1984．5．
18．童海宝．酶催化——催化反应的新领域．精细与专用化学品．1993(13)．6．
19．沈寅初．生物技术在化学工业中的应用．上海化工．1998(12)．35-38．
20．雷武．令人瞩目的生物反应技术．化工之友．1999(4)．26-27．
21．周昕，杨丽等．生物酶在表面活性剂合成及改性中的应用．日用化学工业．1999(3)．60—61．
22．唐平，雕鸿荪．用于油脂改性的1，3-定向脂肪酶．中国油脂．1996．2l(4)．4-9．
23．谷利伟，赵金兰．酶技术在油脂工业中的应用．中国油脂．1998．23(5)．43-45．
24．吴可克，程菊花，高培焕．酶促法合成甘油单酸酯．表面活性剂工业．1996(4)．28-29．
25．黄显慈．选择性合成单双甘油酯．四川化工．1995(4)．52-54．
26．王明．油脂酶催化甘油解制备甘油单酯产率的进一步提高．日用化学工业译丛．1992(1)．19-22．
27．王万绪．油脂的高产率酶催化甘油解．日用化学工业译丛．1992(1)．15-18．
28．彭立风，谭天伟．脂肪酶催化棕榈油甘油解反应合成单硬脂酸甘油酯．北京化工大学学报．1998．25(2)．6-10．
29．彭立风，谭天伟．有机相脂肪酶催化合成单硬脂酸甘油酯的研究进展．化学世界．1998(3)．119-126．
30．罗贵民．有机溶剂中的酶催化反应．精细与专用化学品．1999(5)．12-13．
31．姚传义，俞耀庭等．反胶团酶反应研究进展．化工进展．1999(5)．22-25．
32．刘俊康，刘瑛等．微乳凝胶固定化酶催化长链脂肪酸酯的合成．无锡轻工大学学报．2000．19(4)．361-364．
33．吴姣莲．牛油的固相酶催化甘油解生产高产率甘油单酯．日用化学工业译丛．1991(6)．18-22．
34．阎建民，马润宇．乳膜技术及其在酶化工的应用．化工进展．1999(5)．35-38．