双水相体系萃取精氨酸脱亚胺酶

报道了利用聚乙二醇/硫酸铵双水相体系从自溶NJ402菌粗提酶液中分离纯化精氨酸脱亚氨酶(ADI)的研究结果,为精氨酸脱亚氨酶的分离纯化提供了一种方法。在双水相体系中采用聚乙二醇(PEG)与(NH4)2SO4为组成成分,考察了聚乙二醇(PEG)平均相对分子质量、PEG质量分数、(NH4)2SO4质量分数、pH及NaCl质量分数对精氨酸脱亚氨酶分离纯化效果的影响。最佳双水相体系萃取条件为:聚乙二醇(PEG)平均相对分子质量为1 000,w(PEG1000)=15%,w[(NH4)2SO4]=20%,pH=6.5,室温下从自溶NJ402菌粗提酶液中分离纯化精氨酸脱亚氨酶,纯化倍数达到2.35倍,萃取率达91.1%。     

精氨酸脱亚胺酶发酵条件研究

对一种新型肿瘤增殖抑制酶类一精氨酸脱亚胺酶的发酵生产工艺进行了研究。运用单因子扫描方法,考察了不同碳源、氮源等影响因子对粪肠球菌（Enterococcus faecalis）NJ402产精氨酸脱亚胺酶的影响。优选出其较佳培养基组分为（g／L）;蔗糖15,蛋白胨5,酵母膏5,牛肉膏2．5,NaCl3,KH2PO42,MgSO40．01,MnSO40．0025,L-精氨酸15;接种量以4％～5％为宜,最佳培养温度是37℃,最适起始pH为7．5。100L发酵罐实验表明,发酵到10h时,菌量与比酶活量均为最大,分别为40mg／mL和2．57U／mL。     

反胶束萃取蛋白质技术的新进展

本文综述了近年来反胶束萃取蛋白质这一技术在基础理论研究、应用和过程开发三方面所取得的新进展，指出了某些不足之处，并对今后的研究工作提出了建议。     

肽基精氨酸脱亚胺酶4抗体定性检测试剂盒的研制及初步应用

目的研制肽基精氨酸脱亚胺酶4(Peptidylarginine deiminase 4,PADI4)抗体定性检测ELISA试剂盒,并初步探讨其对诊断类风湿关节炎(Rheumatoid arthritis,RA)的临床意义及应用价值。方法采用PADI4多肽片段作为包被抗原,制备PADI4抗体定性检测试剂盒,并检测50份RA患者和283份健康人血清,评价该试剂盒的分析性能、精密性和稳定性。使用该试剂盒对181份临床确诊RA患者和353份健康人的血清样本进行初步检测。结果所制备的PADI4抗体定性检测试剂盒总体符合率为85.3%,灵敏度为46.0%,特异性为92.2%;板内变异系数分别为2.47%和4.26%,板间变异系数分别为4.24%和6.20%;在4℃保存6个月,总体符合率、灵敏度、特异性、精密性均无明显变化,稳定性良好。181份RA患者血清中阳性检出率为45.3%,353份健康人血清中阳性检出率为7.7%,差异有统计学意义(P<0.01)。结论所研制的试剂盒可用于类风湿关节炎的临床辅助诊断。     

反胶束中酶催化反应研究进展

反胶束具有独特的优良性能,近年来越来越多地受到关注,具有广阔的应用前景.本文介绍了反胶束酶催化理论的最新研究进展,对存在的问题进行了评述,并对其应用前景作了展望.     

反胶束萃取牛血清白蛋白

研究了ＣＴＡＢ－正己醇－正辛烷反胶束溶液萃取ＢＳＡ的性能。考察了水相的ｐＨ值与离子强度和种类、有机相中表面活性剂浓度和助溶剂浓度等因素对萃取行为的影响，并从反胶束的微观结构予以解释。     

离心萃取器用于反胶束溶液萃取牛血清白蛋白

离心萃取器用于反胶束溶液萃取牛血清白蛋白陆强，李宽宏，施亚钧（华东理工大学化学工程研究所，上海２００２３７）关键词离心萃取器，反胶束溶液，牛血清白蛋白１前言以表面活性剂溶于非极性有机溶剂所形成的反胶束溶液为萃取剂，提取与分离水溶液中的蛋白质，是一种有...     

反胶束技术在生物工程中的研究进展

本文综述了反胶束萃取的基础理论研究,反胶束作为酶催化反应介质的优点,酶在反胶束中的催化活性和稳定性以及反胶束技术的应用。     

AOT反胶束萃取胰蛋白酶的STM及主要影响因素

In this article, the influence factors of trypsin extracted from crude pancreatin was investigated, and scanning tunneling microscope(STM) was used to observe the image of trypsin in butane-diacid-2-ethyl-hexyl-ester-sulfonic sodium (AOT)/iso-octane reversed micelles. The STM image showed that trypsins bounded in reversed micelles was rigid, which weakened its conjugative effect and caused maximum ultraviolet absorption and fluorescence emissive absorption moving toward blue waves. AOT concentration, pH and cations were the main influence factors of extraction. Specifically, extraction percentage of trypsin decreased with the increase of AOT concentration from 0.01 to 0.1mol·L-1. When pH value is from 5.30 to 10.0, i.e. less than pI of trypsin, the extraction percentage is raised with the different increase of pI-pH, but when the pH value is less than 5.20, the extraction percentage is decreased with the acidity added. Besides, the extraction efficiency is negative, related with the concentrations of Ca2 , Na ,K which were in the range of 0.2-1.0mol· L-1, and influence of concentration of Ca2 is greater than that of Na , and K which has the minimum impact with the same concentration. Finally, optimum conditions to extract trypsin were: AOT reversed micelles 0.05mol·L-1, trypsin concentration in crude pancreatin solution 3mg·ml-1, pH 5.2- 5.3, ratio (by volume) of extraction phase to strip-extraction phase 1:1, and time of 5min. The corresponding percentage of extraction was 22.7% and specific activity was 78.9 N-benzoyl-L-arginine ethyl ester (BAEE) U·mg-1 protein, three times than that in crude pancreatin. There was no lipase and amylopsin activity was decreased to 1/5 of crude pancreatin. Partly purifying solution was treated by condition mentioned above with 0.05mol·L-1 ceryl-trimethyl-ammonium bromide (CTAB), total extraction percentage of trypsin was 74.18% and specific activity was 3148.3 BAEE U·mg-1, i.e. 48.16 times purer than that in crude pancreatin. Through sodium dodecyl sulfate-polyacryl amide gel electrophoresis (SDS-PAGE) and image analysis of extracted product, there were only three bands in the trypsin, while seven in crude pancreatin, and electrophoresis location of main bend was almost identical with the standard enzyme.     

气溶胶OT反胶束对L-胱氨酸的萃取行为研究

实验考察了气溶胶OT (AOT即 2 乙基己基琥珀酸酯磺酸钠 )浓度、盐酸浓度、L 胱氨酸浓度、无机盐浓度及温度对AOT反胶束萃取L 胱氨酸的影响。结果表明 :AOT浓度增大萃取分配系数增大 ;水相中盐酸浓度增大、无机盐浓度增大、胱氨酸初始浓度增大均使萃取分配系数降低 ;萃取温度升高使萃取的分配系数减小 ,说明AOT反胶束萃取胱氨酸是一个放热过程 ,低温有利于胱氨酸的萃取     

聚合物胶团萃取与聚合物反胶团萃取

介绍了从两亲聚合物及聚合物胶团的形成、聚合物胶团的加溶作用、聚合物胶团加溶理论等方面对聚合物胶团萃取的研究进展做了综述。同时,介绍了作者提出并研究的聚合物反胶团萃取的特性及影响因素。     

反胶团萃取蛋白质设备

综述了反胶团萃取蛋白质设备的研究进展.按设备形式和操作方式的不同将反胶团萃取设备分为膜萃取器、离心萃取器、混合澄清槽和微分萃取器,指出了各自的优缺点,强调了萃取设备的基础研究和开发新型反胶团萃取设备的必要性.     

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。     

Forward and Backward Extraction of Methylene Blue by using AOT/Isooctane Reversed Micellar Solution

Organic dyes, which are contained in industrial effluents, should be removed to avoid health hazards and destruction of the ecosystem. In this study, the extraction of methylene blue from aqueous solution into AOT/isooctane reversed micellar solution was investigated. It was found that methylene blue was solubilized into the waterpool within reversed micelles by electrostatic interaction with AOT. The extraction ratio of methylene blue increased with an increase in AOT concentration and a decrease in salt concentration. The methylene blue extracted reversed micelles could be recovered into fresh salt solution with high concentration. It is considered that the main driving force of forward and backward extraction of methylene blue is electrostatic interaction between cationic dye, methylene blue, and anionic surfactant, AOT. The deterioration of the forward and backward extraction behavior by using AOT/isooctane reversed micellar solution reused was not observed.     

Reversed Micellar Extraction of Vancomycin: Effect of pH,Salt Concentration,and Affinity Ligands

Abstract

Vancomycin was extracted from an aqueous feed solution into a reverse micellar solution of bis(2-ethylhexyl)sulfosuccinate sodium salt in isooctane. A low pH and salt concentration of aqueous feed solutions favors forward extraction. The backward extraction efficiency of vancomycin from reverse micelles into an aqueous phase, on the other hand, increases with pH. Affinity cosurfactants prepared by attaching a dipeptide d-alanyl-d-alanine or a racemic dipeptide dl-alanyl-dl-alanine to cholesteryl-chloroformate was employed for affinity reverse micellar extraction of vancomycin. The forward extraction efficiency increases significantly in the presence of an affinity cosurfactant. The recovery of vancomycin from fermentation broth with high selectivity was also achieved by employing this affinity cosurfactant.     

L-瓜氨酸酶法制备中精氨酸脱亚氨酸酶的活性与稳定性研究

A novel Enterococcus faecalis strain designated N J402 was found with high activity of arginine deiminase （ADI）. The optimum condition for catalytic activity was determined in terms of temperature （about 40℃）, thermostability （available 37℃） and pH （6-7）. The effects of substrate and product concentration were studied. The effects of various metal ions added in reaction mixtures on the biocatalyst were investigated and ADI of N J402 was found to exhibit Co^2＋ dependence, different from previous reports. Surfactant, cetyl trimethyl ammonium bromide, was one of the most important keys for producing L-citrulline. The enzyme in resting cells possessed the quality of high stability for reuse.     

表面活性剂在生物工程中的应用之一—反胶团萃取

反胶团萃取分离蛋白质是一种新型的、有发展前途的生物产品的分离技术。它是利用表面活性剂在有机溶剂中形成反向胶团,从而实现了对蛋白质的萃取,是表面活性剂在生物工程中的一种成功应用。     

固定化细胞酶法拆分DL-精氨酸

以DL-精氨酸为原料,用固定化细胞酶法拆分DL-精氨酸,并对拆分条件进行了研究。结果表明:最适反应温度55℃,最适pH=6.0,c(DL-精氨酸)=0.3 mol/L,ρ(菌体)=30 g/L,拆分反应10 h,拆分率达98%以上。连续反应10批次,固定化细胞仍保留最高酶活力的75%。产物经分离纯化,D-精氨酸收率达84.0%,[α]2D5=-27.4°〔ρ(D-精氨酸)=20 g/L,5 mol/L盐酸〕。