预处理固定化脂肪酶膜对酯化反应的影响
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摘要
考察经多种天然添加剂、活性剂成分及有机相反应前预处理实验室自制固定化酶膜对十二酸十二醇、维生素A/C棕榈酸酯、棕榈酸异辛酯等酯化合成的影响。结果发现:经过明胶、多羟基化合物PVP及含活性剂成分复配乳液PDMS等处理后,酶膜对十二酸十二醇酯化酶活有1.2—1.5倍不同程度提高;相比冻干酶粉完全失活含质量分数98%PEG6000的冻干酶粉催化酯化率在70%以上,结合盐及聚乙二醇高分子化合物对酶保护作用,该方法适于冻干Candida sp.99-125脂肪酶粉的预处理;而在所选常用有机溶剂中酶活性稳定,可用于反应后酶膜的再生。在棕榈酸异辛酯的酯化合成中,复合乳剂PDMS对酶稳定性没有影响,而天然羟基化合物PVP及乳糖明显有利于酶活性的保持、酶重复使用次数的增加。
Immobilized lipase membrane prepared in our lab pre-treated with natural additives and active agent compositions by presoaking in solution,or immersing in organic phase was studied. And the enzymatic character was measured by reaction of lauric acid lauryl ester,vitamin A/C palmitate and ethylhexyl palmitate,as model study of its esterification activity. After natural additives and ingredients emulsion containing surfactant compound treated,an activity increase of 1. 2-1. 5 times was found in lauric acid lauryl ester reaction. In addition,for freezedried lipase powder,the catalytic esterification rate of VA palmitate synthesis was over 70%( freeze-dried enzyme powder mass fraction 98% PEG6000) while the lipase powder without additives having no activity. Considering the protective effect resulting from salt and polyethylene glycol polymers,freeze-dried enzyme powder from Candida sp.99-125 with those additives can be recognized as one kind of pretreatment method. Organic solvents with different hydrophobic constants( almost have no negative effect on lipase activity) can be applied for enzyme regeneration after reaction. Moreover,PVP-360 and lactose are conducive to the stability and using times of the immobilized lipase membrane in esterification of ethylhexyl palmitate.
Immobilized lipase membrane prepared in our lab pre-treated with natural additives and active agent compositions by presoaking in solution,or immersing in organic phase was studied. And the enzymatic character was measured by reaction of lauric acid lauryl ester,vitamin A/C palmitate and ethylhexyl palmitate,as model study of its esterification activity. After natural additives and ingredients emulsion containing surfactant compound treated,an activity increase of 1. 2-1. 5 times was found in lauric acid lauryl ester reaction. In addition,for freezedried lipase powder,the catalytic esterification rate of VA palmitate synthesis was over 70%( freeze-dried enzyme powder mass fraction 98% PEG6000) while the lipase powder without additives having no activity. Considering the protective effect resulting from salt and polyethylene glycol polymers,freeze-dried enzyme powder from Candida sp.99-125 with those additives can be recognized as one kind of pretreatment method. Organic solvents with different hydrophobic constants( almost have no negative effect on lipase activity) can be applied for enzyme regeneration after reaction. Moreover,PVP-360 and lactose are conducive to the stability and using times of the immobilized lipase membrane in esterification of ethylhexyl palmitate.
引文
[1]IYER P V,ANANTHANARAYAN L.Enzyme stability and stabilization-aqueous and non-aqueous environment[J].Process Biochemistry,2008,43(10):1019-1032.
[2]ZHAO Xuebing,QI Feng,YUAN Chongli,et al.Lipasecatalyzed process for biodiesel production:enzyme immobilization,process simulation and optimization[J].Renewable and Sustainable Energy Reviews,2015,44:182-197.
[3]徐岩,李建波.微生物脂肪酶的固定化及其在非水相催化中的应用研究[J].工业微生物,2001,31(1):46-48.
[4]宋宝东,邢爱华,吴金川,等.表面活性剂包衣的固定化脂肪酶在有机介质中催化酯化反应[J].化学工程,2004,32(5):50-52.
[5]张军,徐家立.固定化假丝酵母1619脂肪酶催化油酸油醇酯的合成[J].生物工程学报,1995,11(4):325-331.
[6]李群,袁勤生,李永丰.海藻糖对酶热稳定性保护作用的研究[J].药物生物技术,1997,4(1):28-31.
[7]VRUSHALI Dandavate,DATTA Madamwar.Novel approach for the synthesis of ethyl isovalerate using surfactant coated candida rugosa lipase immobilized in microemulsion based organogels[J].Enzyme Microb Technol,2007,41(3):265-270.
[8]杨立荣,吴坚平,姚善泾.高浓度盐系统中脂肪酶的固定化及其催化活力[J].有机化学,2002,22(3):189-193.
[9]JIN Chuan Wu,LEE S S,MAHMOOD M M B,et al.Enhanced activity and stability of immobilized lipases by treatment with polar solvents prior to lyophilization[J].Journal of Molecular Catalysis B:Enzymatic,2007,45(3/4):108-112.
[10]程媛媛,陈必强,谭天伟.以天然纤维织物为载体固定化脂肪酶[J].北京化工大学学报:自然科学版,2008,35(6):71-74.
[11]FERNNDEZLORENTE G,PALOMO J M,MATEO C,et al.Glutaraldehyde cross-linking of lipases adsorbed on aminated supports in the presence of detergents leads to improved performance[J].Biomacromolecules,2006,7(9):2610-2615.
[12]CHEN Jech-wei,WU Wenteng.Regeneration of immobilized candida antarctica lipase for transesterification[J].J Biosci Bioeng,2003,95(5):466-469.
[13]陈必强,叶华,谭天伟.固定化假丝酵母脂肪酶合成棕榈酸异辛酯[J].化工学报,2004,55(3):422-425.
[14]李宏亮,胡晶,谭天伟.固定化脂肪酶合成维生素A棕榈酸酯[J].生物工程学报,2008,24(5):817-820.
[15]徐凤杰,谭天伟.脂肪酶催化合成异维生素C棕榈酸酯及其动力学[J].北京化工大学学报:自然科学版,2007(S2):95-98.
[16]LI Weina,SHEN Huaqing,TAO Yifeng,et al.Amino silicones finished fabrics for lipase immobilization:Fabrics finishing and catalytic performance of immobilized lipase[J].Process Biochemistry,2014,49(9):1488-1496.
[17]KARTAL F.Enhanced esterification activity through interfacial activation and cross-linked immobilization mechanism of Rhizopus oryzae lipase in a non-aqueous medium[J].Biotechnology Progress,2016,32(4):899-904.
[18]LU Jike,DENG Li,ZHAO Rui,et al.Pretreatment of immobilized Candida sp.99-125 lipase to improve its methanol tolerance for biodiesel production[J].Journal of Molecular Catalysis B Enzymatic,2010,62(1):15-18.
[19]MATEO C,PALOMO J M,FERNANDEZ-LORENTE G,et al.Improvement of enzyme activity,stability and selectivity via immobilization techniques[J].Enzyme Microb Technol,2007,40:1451-1463.
[20]PALOMO J M,MU~NOZ G,FERNNDEZ-LORENTE G,et al.Interfacial adsorption of lipases on very hydrophobic support(octadecyl-Sepabeads):immobilization,hyperactivation and stabilization of the open form of lipases[J].J Mol Catal B:Enzym,2002,19/20:279-286.
[21]BASTIDA A,SABUQUILLO P,ARMISEN P,et al.A single step purification,immobilization,and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports[J].Biotechnol Bioeng,1998,58:486-493.
[22]SHU Zhengyu,JIANG Huan,LIN Ruifeng,et al.Technical methods to improve yield,activity and stability in the development of microbial lipases[J].Journal of Molecular Catalysis B Enzymatic,2010,62(1):1-8.
[23]DARVISHI F,DESTAIN J,NAHVI I,et al.Effect of additives on freeze-drying and storage of Yarrowia lipolytica lipase[J].Applied Biochemistry&Biotechnology,2012,168(5):1101-1107.
[24]FU Dayan,LI Cong,LU Jike,et al.Relationship between thermal inactivation and conformational change of Yarrowia lipolytica lipase and the effect of additives on enzyme stability[J].Journal of Molecular Catalysis B Enzymatic,2010,66(1/2):136-141.
[25]郭诤,张根旺.脂肪酶的结构特征和化学修饰[J].中国油脂,2003,28(7):5-10.
[2]ZHAO Xuebing,QI Feng,YUAN Chongli,et al.Lipasecatalyzed process for biodiesel production:enzyme immobilization,process simulation and optimization[J].Renewable and Sustainable Energy Reviews,2015,44:182-197.
[3]徐岩,李建波.微生物脂肪酶的固定化及其在非水相催化中的应用研究[J].工业微生物,2001,31(1):46-48.
[4]宋宝东,邢爱华,吴金川,等.表面活性剂包衣的固定化脂肪酶在有机介质中催化酯化反应[J].化学工程,2004,32(5):50-52.
[5]张军,徐家立.固定化假丝酵母1619脂肪酶催化油酸油醇酯的合成[J].生物工程学报,1995,11(4):325-331.
[6]李群,袁勤生,李永丰.海藻糖对酶热稳定性保护作用的研究[J].药物生物技术,1997,4(1):28-31.
[7]VRUSHALI Dandavate,DATTA Madamwar.Novel approach for the synthesis of ethyl isovalerate using surfactant coated candida rugosa lipase immobilized in microemulsion based organogels[J].Enzyme Microb Technol,2007,41(3):265-270.
[8]杨立荣,吴坚平,姚善泾.高浓度盐系统中脂肪酶的固定化及其催化活力[J].有机化学,2002,22(3):189-193.
[9]JIN Chuan Wu,LEE S S,MAHMOOD M M B,et al.Enhanced activity and stability of immobilized lipases by treatment with polar solvents prior to lyophilization[J].Journal of Molecular Catalysis B:Enzymatic,2007,45(3/4):108-112.
[10]程媛媛,陈必强,谭天伟.以天然纤维织物为载体固定化脂肪酶[J].北京化工大学学报:自然科学版,2008,35(6):71-74.
[11]FERNNDEZLORENTE G,PALOMO J M,MATEO C,et al.Glutaraldehyde cross-linking of lipases adsorbed on aminated supports in the presence of detergents leads to improved performance[J].Biomacromolecules,2006,7(9):2610-2615.
[12]CHEN Jech-wei,WU Wenteng.Regeneration of immobilized candida antarctica lipase for transesterification[J].J Biosci Bioeng,2003,95(5):466-469.
[13]陈必强,叶华,谭天伟.固定化假丝酵母脂肪酶合成棕榈酸异辛酯[J].化工学报,2004,55(3):422-425.
[14]李宏亮,胡晶,谭天伟.固定化脂肪酶合成维生素A棕榈酸酯[J].生物工程学报,2008,24(5):817-820.
[15]徐凤杰,谭天伟.脂肪酶催化合成异维生素C棕榈酸酯及其动力学[J].北京化工大学学报:自然科学版,2007(S2):95-98.
[16]LI Weina,SHEN Huaqing,TAO Yifeng,et al.Amino silicones finished fabrics for lipase immobilization:Fabrics finishing and catalytic performance of immobilized lipase[J].Process Biochemistry,2014,49(9):1488-1496.
[17]KARTAL F.Enhanced esterification activity through interfacial activation and cross-linked immobilization mechanism of Rhizopus oryzae lipase in a non-aqueous medium[J].Biotechnology Progress,2016,32(4):899-904.
[18]LU Jike,DENG Li,ZHAO Rui,et al.Pretreatment of immobilized Candida sp.99-125 lipase to improve its methanol tolerance for biodiesel production[J].Journal of Molecular Catalysis B Enzymatic,2010,62(1):15-18.
[19]MATEO C,PALOMO J M,FERNANDEZ-LORENTE G,et al.Improvement of enzyme activity,stability and selectivity via immobilization techniques[J].Enzyme Microb Technol,2007,40:1451-1463.
[20]PALOMO J M,MU~NOZ G,FERNNDEZ-LORENTE G,et al.Interfacial adsorption of lipases on very hydrophobic support(octadecyl-Sepabeads):immobilization,hyperactivation and stabilization of the open form of lipases[J].J Mol Catal B:Enzym,2002,19/20:279-286.
[21]BASTIDA A,SABUQUILLO P,ARMISEN P,et al.A single step purification,immobilization,and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports[J].Biotechnol Bioeng,1998,58:486-493.
[22]SHU Zhengyu,JIANG Huan,LIN Ruifeng,et al.Technical methods to improve yield,activity and stability in the development of microbial lipases[J].Journal of Molecular Catalysis B Enzymatic,2010,62(1):1-8.
[23]DARVISHI F,DESTAIN J,NAHVI I,et al.Effect of additives on freeze-drying and storage of Yarrowia lipolytica lipase[J].Applied Biochemistry&Biotechnology,2012,168(5):1101-1107.
[24]FU Dayan,LI Cong,LU Jike,et al.Relationship between thermal inactivation and conformational change of Yarrowia lipolytica lipase and the effect of additives on enzyme stability[J].Journal of Molecular Catalysis B Enzymatic,2010,66(1/2):136-141.
[25]郭诤,张根旺.脂肪酶的结构特征和化学修饰[J].中国油脂,2003,28(7):5-10.