CTAB混合反胶团萃取工业脂肪酶

研究了CTAB与Tween、含氧有机物形成的混合反胶团对工业脂肪酶进行萃取分离的效果. 实验表明,CTAB-Tween85和CTAB-含氧有机物混合反胶团的萃取率高于单一CTAB反胶团;反萃时CTAB-含氧有机物混合反胶团的反萃率与单一CTAB反胶团的反萃率相似,CTAB-Tween60和Tween40混合反胶团的反萃效果优于单一CTAB反胶团. 通过测定反萃水相的酶活,发现CTAB-TRPO混合反胶团的效果最好,酶活回收率最高,可以达到70%.     

AOT-磷脂反胶团体系萃取蛋白质的研究

研究了PC/AOT和PE/AOT混合体系在有机溶剂中形成反胶团的性质及其萃取蛋白质的性能,并解决了反胶团萃取中有磷脂存在时的蛋白质浓度的分析问题.研究结果表明,在AOT体系中加入磷脂能使胶团尺寸变大,且加入PE能提高血红蛋白和枯草杆菌a—淀粉酶的萃取率,加入PC则常使萃取性能变差.因此,在反胶团萃取蛋白质时,影响萃取的主要因素不仅是胶团的大小,还有胶团与蛋白质之间的相互作用.     

反胶团萃取蛋白质特性的研究

实验将一种阴离子表面活性剂（AOT）和一种阳离子表面活性剂（Aliquat336）分别溶于异辛烷（isooctane）中，构成了两种不同的反胶团体系．通过用两种不同的反胶团溶液萃取六种蛋白质的实验，研究了水相pH值及离子强度对反胶团体系中水含量Wo和蛋白质萃取率影响的规律．实验发现不论哪种反胶团体系，水相pH值对Wo的影响都不大，但pH值却对蛋白质萃取率有很大的影响，对AOT反胶团体系而言，随着pH值的降低，蛋白质的萃取率升高；对Aliquat336反胶团体系而言，随着水相pH值的降低，蛋白质的萃取率下降．随着离子强度的增大，AOT反胶团体系的水含量大幅度地降低；而对Aliquat336体系水含量的影响很小，但水相离子强度对蛋白质萃取率的影响是相同的，随着离子强度的增大，两种体系对蛋白质的萃取率均下降。     

正交实验优化超临界流体CO_2反胶团-络合萃取痕量重金属

利用超临界流体CO2反胶团-络合萃取食品中痕量重金属铅、汞和砷,通过正交实验考察了萃取压力、萃取温度、萃取时间和反胶团AOT浓度对痕量重金属萃取率的影响。确定最佳萃取工艺为:萃取压力20 MPa、萃取温度45℃、萃取时间90 min、反胶团AOT浓度0.1 mol.L-1,此时,重金属铅、汞、砷的萃取率分别为93.50%、95.36%、90.47%。表明超临界流体CO2络合萃取时加入反胶团能明显提高痕量重金属的脱除率,萃取效果优于超临界流体CO2反胶团萃取和超临界流体CO2络合萃取。     

基于表面活性剂的反胶团萃取技术

基于表面活性剂的反胶团萃取技术是一种新型的有发展前途的生物分离技术。介绍了反胶团萃取技术的驱动力、表面活性剂选择和影响因素,提出了反胶团萃取技术目前存在的问题,探讨了反胶团萃取技术在其酶促反应和纳米材料制备方面的应用及其发展方向。     

新型混合反胶团萃取溶菌酶的平衡行为

研究了由阴离子表面活性剂二-(2-乙基己基)琥珀酸酯磺酸钠(AOT)和非离子表面活性剂聚氧乙烯基(聚合度为4)壬基酚醚(OPE4)组成的一种新型混合反胶团体系及其萃取溶菌酶的平衡行为. 结果表明,该反胶团体系具有较大的含水量,且其含水量在较大的表面活性剂配比(0~0.8)范围内维持恒定,是由静电作用和胶束形态改变共同作用的结果. 无机盐种类和离子强度对上述混合反胶团的含水量有着显著的影响,继而影响到溶菌酶的萃取率. 它可归结为盐离子对扩散双电层和水化膜斥力的因素所致. 降低pH和提高总表面活性剂浓度均有利于溶菌酶的萃取.     

反胶团系统及蛋白质萃取过程研究进展

综述了反胶团系统和蛋白质萃取过程,将反胶团萃取系统按单一反胶团系统、混合反胶团系统和亲和反胶团系统划分,强调了研究开发生物相容性表面活性剂以及在反胶团系统中引入亲和作用的重要性。另外,指出了深入开展反胶团萃取设备和过程研究的必要性。     

复合稳定剂对碱性蛋白酶的作用研究

通过单因素实验得出了对碱性蛋白酶稳定效果较好的几种保护剂:5mmol·L-1 Ca2 ;20mg·mL-1微肢粒抑制剂丙二醇-单甲醚;0.01%(质量体积比)可逆蛋白酶活性抑制剂4-甲酰苯基硼酸;1%甘油.在此基础上,进行L18(37)正交实验,以相对酶活率为指标,考察了不同酶活性保护荆在不同条件下对液体碱性蛋白酶活力的影响,筛选到一种优质高效的碱性蛋白酶稳定荆配方为:5mmol·L-1Ca2 ,15mg·mL-1丙二醇-单甲醚,0.015%4-甲酰苯基硼酸,1%甘油.加入液体碱性蛋白酶40℃保存15d后,相对酶活力仍保持在85%以上.     

反胶团酶催化研究新进展

评述了近年来反胶团酶催化研究的新进展。在AOT/异辛烷反胶团中加入非离子型表面活性剂如Tween 85、小相对分子质量聚乙二醇等可有效降低酶与表面活性剂间的静电和疏水作用 ,显著提高酶的活性。对AOT进行化学修饰及合成结构与磷脂类似的新表面活性剂以用其构建新的反胶团体系 ,酶的活性较常用的AOT/异辛烷反胶团体系有显著提高。在反胶团酶反应动力学研究中考虑水含量或底物在反胶团表面吸附的影响等 ,提出了进一步研究的设想 ,包括开发新型表面活性剂以进一步提高酶的活性和稳定性及有利于产物分离     

阳离子反胶团体系萃取牛血清白蛋白

采用一种新的阳离子反胶团体系(CTAB/异辛烷-正戊醇),研究了反胶团萃取牛血清白蛋白(BSA).用单变量法考察了表面活性剂浓度、水相pH值、离子种类和浓度、萃取时搅拌速率和油水比对BSA萃取率的影响.实验表明,静电作用力是该萃取过程的主要动力,随水相pH值增大,BSA的萃取率逐渐升高;对于不同种类的离子,基本随其浓度增大,萃取率下降.在优化的操作条件下,BSA萃取率可达98%以上,因此,CTAB/异辛烷-正戊醇反胶团体系适合BSA的萃取.     

Activity of horseradish peroxidase in aqueous and reverse micelles and back‐extraction from reverse‐micellar phases

Studies on the activity of the enzyme horseradish peroxidase (HRP) have been carried out in micellar as well as reverse‐micellar media. The activity of the enzyme was studied in the presence of different classes of surfactants – ionic as well as non‐ionic. In aqueous media, the activity of the enzyme varied depending on whether the concentration of the surfactant used was above or below the critical micellar concentration (CMC). The enzyme was also studied in reverse‐micellar systems. HRP was introduced into the reverse micellar phase by the injection method and its activity within the reverse micelles was determined. The effect of water to surfactant ratio (W_o) on activity within reverse micelles was studied, and an almost two‐fold increase in activity was seen when the enzyme was encapsulated within reverse micelles of aqueous phase fractional hold‐up (?) of 0.0072 (v/v) consisting of sodium bis‐(2‐ethylhexyl) sulfosuccinate (AOT) in isooctane at a W_o of 20. The activity of HRP was measured over a wide range of AOT concentrations having different W_o values. Back‐extraction of HRP from these reverse micelles was carried out at varying ionic strengths of phosphate buffer. Back extraction was found to be highest at pH 7.0 in 40 mol m^?3 phosphate buffer and 100 mol m^?3 sodium chloride. © 2001 Society of Chemical Industry     

反胶束体系中酶法合成头孢力新的动力学

利用来自Xanthomonas citri的酰化酶由D-α-苯甘氨酸甲酯和7-ADCA合成头孢力新是典型的动力学控制合成过程,其中既有产物的合成又有产物和底物（D-α-苯甘氨酸甲酯）的水解。动力学分析表明,反应置于反胶束体系将不仅提高酶的催化速度而且提高转移酶活力对水解酶活力的选择性。利用AOT/异辛烷反胶束体系酶法合成头孢力新使水解副反应被抑制,合成途径被强化。水成为反胶束酶反应体系中重要的调节手段,它对反应速度呈“铃形”关系。     

Enzymatic esterification of DL‐menthol with propionic acid by lipase from Candida cylindracea

This work deals with the resolution of DL ‐menthol with propionic acid by Candida cylindracea lipase (Ccl) in organic solvent reaction systems and a reverse micelles system of sodium 1,4‐bis (2‐ethylhexyl) sulfosuccinate (AOT). The activity and stability as well as enantioselectivity of the lipase in two systems were studied. The results indicate that the lipase showed higher stability in reverse micelles than in organic solvent, which proved that the reverse micelles system has potential application for maintaining the activity of the enzyme for a long time. This is because lipase molecules can be entrapped in water‐containing micro‐drops of reverse micelles, avoiding direct‐contract with unfavorable organic medium. The enantioselectivity (E > 30, ee_p = 92.5) in the two systems is relatively high, although the conversion is moderate. The influence of the characteristic parameters of the two systems, such as pH, temperature, w₀ (molar ratio of water to AOT in reverse micelles systems) and water content (organic solvent) on the conversion of DL ‐menthol was also investigated. Copyright © 2005 Society of Chemical Industry     

Kinetic behavior of yeast alcohol dehydrogenase in AOT/isooctane reverse micelles

The kinetic behavior of yeast alcohol dehydrogenase (YADH) in sodium di-2-ethylhexylsulfosuccinate (AOT) isooctane reverse micelles has been studied using methyl ethyl ketone (MEK), NADH and Tris as the substrate, coenzyme and buffer, respectively. The solubility diagrams of aqueous buffer in the isooctane solution of AOT were established as a function of temperature and molar ratio of water of surfactant (ω₀) at various Tris and AOT concentrations. The dependence of enzyme activity on enzyme concentration, pH, ω₀ and Tris concentration was determined. The optimal ω₀ was at 10–15, increasing slightly with an increase in Tris concentration. The YADH entrapped in reverse micelles exhibited minimum activity at a Tris concentration of 0·1 mol dm^?3, while in aqueous buffer enzyme activity was not significantly affected by Tris concentration. Comparing the rate equation of the reduction of MEK by YADH in reverse micelles with that in aqueous buffer, the association of YADH and NADH could apparently have proceeded with an irreversible reaction before the substrate was bound, when performed in a reverse micellar system. Although the YADH entrapped in reverse micelles was less stable than when dissolved in aqueous buffer, the enzyme retained at least 20% activity after 21 h at 25°C and ω₀ = 20. This result was an improvement over previously reported data.     

Increased activity and stability of Candida rugosa lipase in reverse micelles formed by chemically modified AOT in isooctane

Sodium bis (2‐ethylhexyl polyoxyethylene) sulfosuccinates, which can be structurally viewed as chemically modified AOT (MAOT), were prepared and successfully used to form reverse micelles in isooctane. The activity and stability of Candida rugosa lipase (glycerol‐ester hydrolase, EC 3.1.1.3) appreciably increased in the MAOT/isooctane reverse micellar system compared with the AOT/isooctane system taking the hydrolysis of olive oil as a model reaction. The best polymerization number of oxyethylene group was found to be 2.0, which corresponded to a maximum enzyme activity of twice that in the AOT/isooctane system. The half‐life of lipase in the MAOT micellar system was twice that in the AOT micellar system. The optimal operational conditions remained unchanged. The Michaelis kinetics showed that the maximum reaction rate obviously increased in the MAOT micellar system compared with that in the AOT micellar system, while the change in the Michaelis constant was insignificant. © 2001 Society of Chemical Industry     

XRD,SEM, and XPS Analysis of Soybean Protein Powders Obtained Through Extraction Involving Reverse Micelles

Soybean protein powders obtained by aqueous buffer and reverse micelle extractions were examined and characterized using X‐ray photoelectron spectroscopy (XPS), X‐ray diffraction (XRD), and scanning electron microscopy (SEM). These analysis methods provided detailed information about elemental distributions, surface structure, and secondary and microstructures of the protein, respectively. XPS data revealed that the O and N atomic percentages of soybean protein surfaces obtained with bis(2‐ethylhexyl) sodium sulfosuccinate (AOT)/hexane reverse micelles were higher than those obtained with aqueous buffer, whereas the percentage of atomic C was lower. The ratios of nitrogen to carbon (N/C) on the surface of soybean protein obtained through the two extraction methods were similar. The O/C ratio for soybean protein obtained from AOT reverse micelles was large. The obtained results indicated that the reverse micelles could affect the C, O, and N components on the surface of soybean proteins. Moreover, XRD and SEM results also showed the influence of AOT reverse micelles, which lead to more β‐sheet and pore structures.     

反胶束中中性蛋白酶AS1.398反应机理

枯草杆菌中性蛋白酶AS1.398可用于反胶束萃取全脂豆粉分离大豆蛋白和油脂.揭示酶的催化作用机理可用于优化萃取条件.以大豆分离蛋白为底物,研究了AS1.398在AOT/异辛烷反胶束体系中的反应机理. 由于底物分子在反胶束中呈泊松分布,通过计算底物和酶分子数比例,推算出酶催化为单底物反应的概率占绝对优势.借鉴水相酶反应模型,考虑反胶束中分子交换作用,提出了反胶束中酶的单底物分子反应的作用模型,模型推导出水相的米氏常数比反胶束相的大1个数量级.实验测定酶在水相和反胶束相的米氏常数分别为3.2×10^-3g•ml^-1和4.2×10^-4g•ml^-1.两者比值与模型推导出的关系较吻合,表明此模型可用来表示反胶束中AS1.398的催化反应.     

Increased activity of Chromobacterium viscosum lipase in AOT reverse micelles in the presence of short chain methoxypolyethylene glycol

The activity of Chromobacterium viscosum lipase (glycerol‐ester hydrolase, EC 3.1.1.3) entrapped in AOT/isooctane and AOT/Tween 85/isooctane reverse micelles was significantly increased by the addition of short chain methoxypolyethylene glycols (MPEGs), taking the hydrolysis of olive oil as a model reaction. The molecular weight of MPEG had a strong effect on the lipase activity, and MPEG of nominal molecular weight 550 was found to be the most effective. To optimize the factors affecting enzymatic hydrolysis of olive oil in reverse micellar systems containing MPEG 550, the effect of various parameters, such as W_o (molar ratio of water to surfactant), pH, ionic strength, surfactant concentration and temperature were investigated. A kinetic model considering the substrate adsorption equilibrium between the bulk phase of organic solvent and the micellar phase was also successfully used to understand the enzyme activity in the presence of MPEG 550. Both the Michaelis constant and the substrate adsorption equilibrium constant were obviously reduced as compared with those obtained in the simple AOT reverse micellar system. © 2001 Society of Chemical Industry