反胶团萃取蛋白质的研究

本文以溶菌酶,胰蛋白酶和胃蛋白酶为对象,研究了水相pH值,离子强度、阳离子种类和蛋白质分子量等因素对反胶团萃取蛋白质的影响。结果表明,反胶团萃取的单级萃取率高,调节PH值和离子强度等工艺条件,就可以实现不同种类蛋白质的有效分离,可望成为一种生物产品分离的新方法。     

反向微胶团萃取分离酪氨酸的研究

研究了AOT反向微胶团的性质 ,并以酪氨酸为反向微胶团的萃取对象 ,探讨了各种因素对酪氨酸分配比的影响 ,优化了萃取、反萃取的工艺条件。     

TRPO-AOT 反胶团体系萃取牛血红蛋白的研究

反胶团是表面活性剂溶解在非极性溶剂中形成的、围绕一个“水核”的纳米级聚集体。液液反胶团萃取蛋白质技术,因对目标物质选择性好、容量大和能保持其活性而得到广泛研究^[1-9].在反胶团萃取蛋白质的研究中,多数作者采用单一表面活性剂AOT^[2]或季胺盐^[3]的反胶团体系。两种体系的共同弱点是:体系受离子强度、pH值等静电因素的影响大,直接影响萃取率,为了克服它们的不足,有人在AOT体系中加亲和试剂增强反胶团对蛋白质的亲和性^[4],加磷酸类阴离子表面活性剂^[5]、天然表面活性剂磷脂^[6]等以增强体系的萃取性能e人在季胺盐的反胶团体系中加非离子表面活性剂作助剂提高蛋白质的萃取率^[7],有人则反阴、阳和非离子表面活性剂混合形成反胶团提高某种酶的萃取容量^[8],本文用中性磷氧萃取剂三烷基氧膦（TRPO）与阴离子表面活性剂琥珀二辛酯磺酸钠（AOT）混合溶解在异辛烷中形成反胶团萃取牛血红蛋白（BHb）,比较AOT、TRPO及AOT三体系对牛血红蛋白（BHb）的萃取性能。     

猪心细胞色素C的反胶团萃取研究

本文研究了AOT/异辛烷对猪心CytC的萃取,分析了不同AOT浓度、pH值条件、体积比、阳离子种类以及离子浓度对反胶团萃取猪心CytC的影响,结果表明;AOT浓度增加,对CytC的萃取率提高;在pH6～9之间对CytC有较好的萃取效果;Mg~(2 )、Na~ 、K~ 、Ca~(2 )”、Mn~(2 )、Li~ 离子中以Li~ 和Mg~(2 )的效果最好,其顺序为:Li~ >Mg~(2 )>Na~ >Ca~(2 )>K~ ,而 Mg~(2 )在0.025mol/L时萃取率最好,可达96.5％;当油:水体积比增大时,萃取率也增大。     

反胶团萃取

反胶团萃取分离技术是一种新型的 ,有发展前途的生物产品分离技术。本文着重对反胶团的表面活性剂 ,各种影响因素 (W0 、pH、T等 ) ,动力学和热力学的理论研究以及目前的开发应用状况等多方面的现状进行综述 ,并对今后的发展进行展望     

生物物质的分离新技术--反胶团萃取

反胶团萃取分离技术是一种新型的,有发展前途的生物产品分离技术。本文着重对反胶团的表面活性剂,各种影响因素(W0、pH、T等),动力学和热力学的理论研究以及目前的开发应用状况等多方面的现状进行综述,并对今后的发展进行展望。     

白地霉脂肪酶的双水相萃取和反胶团提取

对影响双水相萃取和反胶团提取脂肪酶的各种因素进行了探讨,并通过正交实验进一步优化提取条件,PEG浓度15%,(NH4)2SO4浓度22.5%,pH8.0的条件下进行双水相萃取,脂肪酶纯化倍数达到7.5倍;CTAB浓度150mmol/L,相体积比4/2,水相pH8.0,温度40℃的条件下进行反胶团提取,脂肪酶的比活力达到最大,但其比活力稍有下降,约为原来的0.9倍。     

反胶团萃取蛋白质技术的反萃过程研究进展

反胶团萃取分离技术是一种新型的生物产品分离技术,本文简要介绍了反胶团萃取蛋白质技术的反萃过程的动力学,重点综述了在提高蛋白质反萃效率方面的研究进展,并对目前存在的问题、发展方向等进行了评述。     

生物物质的分离新技术--反胶团萃取

反胶团萃取分离技术是一种新型的,有发展前途的生物产品的分离技术。本文着重对反胶团的各种影响因素(表面活性剂的种类、水与表面活性剂的摩尔比WO、pH值、温度T等),动力学和热力学的理论研究以及目前的开发应用状况等多方面的现状进行综述,并对今后的发展进行展望。     

蛋白质的反胶团萃取机制及动力学模型研究进展

反胶团萃取是近年发展起来的分离和纯化生化物质的新方法,本文介绍了反胶团萃取蛋白质技术的原理和机制、影响反胶团中蛋白质稳定性的因素,改进的蛋白质反萃取工艺,反胶团的酶动力学研究以及反胶团萃取技术的研究展望。     

反胶束萃取胰蛋白酶的研究

本文以含有反胶束的有机溶剂作为萃取剂,进行了将胰蛋白酶从水相传入有机相,再从有机相传入另一水相的研究。结果表明：影响萃取率的主要因素为水相ｐＨ值、离子强度和种类,以及反胶束溶液中表面活性剂浓度等;在适宜的条件下,酶的单级萃取和反萃取率都很高,显示了良好的工业应用前景。     

The Lipase/Lipoxygenase Bienzyme System in AOT Reversed Micelles in Octane

In this work it is shown that the bienzyme lipase/lipoxygenase system can function in reversed micelles of bis(2-ethyl)hexyl sulfosuccinate (AOT) in octane. As a lipase substrate, a fish fat preparation (fat of sea mammals) with a high content of polyunsaturated fatty acids was used. It was demonstrated that the bienzyme reaction proceeded in a stationary mode and had a rate-limiting step catalyzed by lipase. Under optimal conditions, the efficacy of functioning of the bienzyme system was by an order of magnitude higher than that in water. The lipase/lipoxygenase bienzyme system can be used as a new method of spectrophotometrical determination of lipase activity.     

微生物转谷氨酰胺酶(MTGase)的AOT反胶束纯化研究

研究微生物转谷氨酰胺酶(MTGase)反胶束纯化的工艺和条件,调节MTGase离心上清液等电点,除去部分杂蛋白,MTGase活力升高7.5倍;用截留分子量为10000的超滤膜除去小分子杂蛋白,MTGase活力升高1.33倍;用0.05mol/L的AOT/异辛烷反胶束进一步纯化MTGase,其最适萃取条件是粗MTGase蛋白质浓度20mg/mL,[Na ]0.12mol/L,水相pH4.80～5.20,相比1:1(v/v);荷载MTGase的AOT反胶束用2.0mol/LKCl进行反萃取,MTGase活力为14.2U/g,纯化8.875倍;冷冻干燥脱盐反萃取液,获得MTGase冻干粉,其活力为110.3U/g,与粗酶液相比较,纯化689.4倍。经过AOT/异辛烷反胶束萃取纯化的MTGase,其SDS-聚丙烯酰胺凝胶电泳为一条带。Ca2 与表面活性剂非极性尾上丁二酰羰基氧、极性头磺酸基硫氧基氧及MTGase分子表面具有孤电子对的基团的配位结合放大了AOT反胶束的另一种萃取作用——配位萃取,致使其对MTGase的萃取率高于K 而接近Na 。     

脂肪酶在反相胶囊中的催化行为的研究

系统研究了脂肪酶在ＡＯＴ／水／异辛烷反应相胶囊中的催化行为。在一定条件下,反相胶囊中的酶反应的仍符合Ｍｉｃｈａｅｌｉｓ－Ｍｅｎｔｅｎ动力学原理,研究了含水量,底物浓度,ｐＨ,温度,溶剂的种类和表面活性剂浓度等对酶反应的影响。结果表明,酶活力与Ｒ值（水与表面活性剂的摩尔比值）有关。获得最大酶活力的条件是Ｒ＝１１,ｐＨ７．０,温度３２．５℃,橄榄油浓度为４０％。     

苦参碱的反胶束萃取研究

本文探索AOT-异辛烷反胶束萃取苦参碱的最佳工艺和条件,以AOT-异辛烷反胶束对粗苦参碱中的苦参碱进行萃取,利用毛细管电泳对苦参碱进行定量分析,通过正交试验优化萃取工艺和条件,确定影响萃取率的主要因素为萃取水相的酸度,次要因素为水相的温度,反胶束的W0和离子强度对萃取率的影响较小,得出最优萃取条件为:pH=12,W0=30,T=35℃。     

Chloroperoxidase-Catalysed Halogenation of Apolar Compounds Using Reversed Micelles

The chloroperoxidase from the mold Caldariomyces fumago was entrapped into reversed micelles composed of aqueous buffer, cetyltrimethylammonium chloride or bromide, pentanol and octane. The surfactant serves a dual function: i) it stabilizes the reversed micelle, and ii) the halide counter ion is used as a substrate for the enzyme. 2-Monochlorodimedon and 1,3-dihydroxybenzene were halogenated with this system, giving their 2-halo and 4-halo derivatives, respectively. The reaction rates were about twice as high as in aqueous media. Enzyme activity was maximal at high water content of the micelles and at relatively low pentanol concentration. The enzyme was inactivated by high concentrations of hydrogen peroxide.     

Chloroperoxidase-Catalysed Halogenation of Apolar Compounds Using Reversed Micelles

The chloroperoxidase from the mold Caldariomyces fumago was entrapped into reversed micelles composed of aqueous buffer, cetyltrimethylammonium chloride or bromide, pentanol and octane. The surfactant serves a dual function: i) it stabilizes the reversed micelle, and ii) the halide counter ion is used as a substrate for the enzyme. 2-Monochlorodimedon and 1,3-dihydroxybenzene were halogenated with this system, giving their 2-halo and 4-halo derivatives, respectively. The reaction rates were about twice as high as in aqueous media. Enzyme activity was maximal at high water content of the micelles and at relatively low pentanol concentration. The enzyme was inactivated by high concentrations of hydrogen peroxide.     

Characteristics of Lipase-Catalyzed Glycerolysis of Triglyceride in AOT-Isooctane Reversed Micelles

Chromobacterium viscosum lipase, solubilized in microemulsion droplets of glycerol containing small amounts of water and stabilized by a surfactant, could catalyze the glycerolysis of triolein. Kinetic analysis of the lipase-catalyzed reaction was possible in the reversed micellar system. Among surfactants and organic solvents tested, bis(2-ethylhexyl)sodiumsulfosuccinate (AOT) and isooctane were respectively most effective, for the glycerolysis of triolein in reversed micelles. Temperature effects, pH profile, K_m,app, and V_max,app were determined. Among various chemical compounds, Fe³⁺, Cu²⁺, and Hg²⁺ inhibited the lipase-catalyzed glycerolysis severely. However, the glycerolysis activity was partially restorable by adding histidine or glycine to the system containing these metal ions. The glycerolysis activity was dependent on water content and maximum activity was obtained at an R value of 1.21. Higher stability of the lipase was obtained in the reversed micellar system.     

Characteristics of Lipase-Catalyzed Glycerolysis of Triglyceride in AOT-Isooctane Reversed Micelles

Chromobacterium viscosum lipase, solubilized in microemulsion droplets of glycerol containing small amounts of water and stabilized by a surfactant, could catalyze the glycerolysis of triolein. Kinetic analysis of the lipase-catalyzed reaction was possible in the reversed micellar system. Among surfactants and organic solvents tested, bis(2-ethylhexyl)sodiumsulfosuccinate (AOT) and isooctane were respectively most effective, for the glycerolysis of triolein in reversed micelles. Temperature effects, pH profile, K_m,app, and V_max,app were determined. Among various chemical compounds, Fe³⁺, Cu²⁺, and Hg²⁺ inhibited the lipase-catalyzed glycerolysis severely. However, the glycerolysis activity was partially restorable by adding histidine or glycine to the system containing these metal ions. The glycerolysis activity was dependent on water content and maximum activity was obtained at an R value of 1.21. Higher stability of the lipase was obtained in the reversed micellar system.