制备了交联血管紧张素转化酶聚集体(ACE-CLEAs),比较了ACE-CLEAs及游离ACE的酶学性质,包括最适酶促反应温度、最适pH值、Km、Vmax、温度稳定性及pH稳定性等.以酶活力回收率为参考,确定了制备ACE-CLEAs的最佳条件为:饱和度为80％的(NH4)2SO4溶液作为沉淀剂,沉淀时间0.5h,质量分数为0.02％的戊二醛作为交联剂,交联时间lh.通过比较酶学性质发现,ACE-CLEAs比游离ACE具有更好的温度稳定性及pH稳定性,且与游离ACE接近的Km值表明,ACE-CLEAs对底物的亲和力与游离酶几乎相当.
参考文献
| [1] | Yamamoto N,Akino A,Takano T.Antihypertensive Effects of Different Kinds of Fermented Milk in Spontaneously Hypertendive Rats[J].Biosci Biotech Biochem,1994,58(4):776-778. |
| [2] | HONG Yunjia,LI Tanyao,CHEN Bo.Immobilization of Angiotensin Converting-enzyme on Chitosan Microspheres[J].Chem J Chinese Univ,2009,30(2):328-331 (in Chinese).洪韫嘉,李谭瑶,陈波.壳聚糖固定化血管紧张素转化酶及其性质[J].高等学校化学学报,2009,30(2):328-331. |
| [3] | LIU Hong,CHEN Lanying.Immobilization and Properties of Angiotensin Converting Enzyme[J].Acta Acad Med Sin,2000,22(6):558-561(in Chinese).刘宏,陈兰英.血管紧张素转化酶的固相化与性质[J].中国医学科学院学报,2000,22(6):558-561. |
| [4] | Schoevaart R,Wolbers M W,Golubovic M,et al.Preparation,Optimization,and Stuctures of Cross-linked Enzyme Aggregates(CLEAs)[J].Biotechnol Bioeng,2004,87(6):754-762. |
| [5] | López-Serrano P,Cao L Q,Rantwijk F V,et al.Cross-linked Enzyme Aggregates with Enhanced Activity:Application to Lipases[J].Biotechnol Lett,2002,24:1379-1383. |
| [6] | Matijosyté I,Arends I W C E,Vries S D,et al.Preparation and Use of Cross-linked Enzyme Aggregates(CLEAs) of Laccases[J].J Mol Catal B:Enzym,2010,62:142-148. |
| [7] | Aytar B S,Bakir U.Preparation of Cross-linked Tyrosinase Aggregates[J].Process Biochem,2008,43:125-131. |
| [8] | DONG Xiaoyi,XIA Shiwen.The Preparation and Application of Cross-linked Urease Aggregates[J].Chinese J Biotechnol,2003,19(3):332-336(in Chinese).董晓毅,夏仕文.交联脲酶聚集体的制备和初步应用[J].生物工程学报,2003,19(3):332-336. |
| [9] | Gallego F L,Betancor L,Hidalgo A,et al.Co-aggregation of Enzymes and Polyethyleneimine:A Simple Method to Prepare Stable and Immobilized Derivatives of Glutaryl Acylase[J].Biomacromolecules,2005,6:1839-1842. |
| [10] | Sheldon R A.Cross-linked Enzyme Aggregates(CLEAs):Stable and Recyclable Biocatalysts[J].Biochem Soc Trans,2007,35 (6):1583-1587. |
| [11] | Toral A R,DelosRíos A P,Hernández F J,et al.Cross-linked Candida Antarctica Lipase B is Active in Denaturing Ionic Liquids[J].Enzyme Microb Teclnol,2007:40:1095-1099. |
| [12] | Wang M F,Qi W,Yu Q X,et al.Cross-linking Enzyme Aggregates in the Macropores of Silica Gel:A Practical and Efficient Method for Enzyme Stabilization[J].Biochem Eng,2010,52:168-174. |
| [13] | WU Xianshan,HE Liqian,YE Lei.Cross-linked Enzyme Aggregates-A Method of Immobilized Enzymes Without Carriers[J].Biotechnol,2005,15(2):90-92(in Chinese).武仙山,何立千,叶磊.交联酶聚集体——种无载体酶固定化方法[J].生物技术,2005,15(2):90-92. |
| [14] | WEI Jiaqian,ZHOU Jianping,ZHANG Wenqi.Progress on the Study of Immobilized Enzymes Without Carriers[J].ansu Sci,2006,18(1):66-70(in Chinese).魏甲乾,周剑平,张文齐.无载体固定化酶的研究进展[J].甘肃科学学报,2006,18(1):66-70. |
| [15] | Ehlers M R W,Maeder D L,Kirsch R E.Rapid Affinity Chromatographic Purification of Human Lung and Kidney Angiotensin Converting Enzyme with the Novel N-Carboxyalkyl Dipeptide Inhibitor N-[l (S)-Carboxy-5-aminopentyi]glycylglycine[J].Biochem Biophys,1986,883:361-372. |
| [16] | Dong T,Zhao L,Huang Y,et al.Preparation of Cross-linked Aggregates of Aminoacylnse from Aspergillus Melleus by Using Bovine Serum Albumin as an Inert Additive[J].Bioresour Technol,2010,101:6569-6571. |
| [17] | Migneault I,Dartiguenave C,Bertrand M J,et al.Glutaraldehyde:Behavior in Aqueous Solution,Reaction with Proteins,and Application to Enzyme Crosslinking[J].Biotechniques,2004,37(5):790-802. |
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