Purification and properties of β-galactosidase from Tilapia intestine: Digestive enzyme of Tilapia-X

ABSTRACT:   β-galactosidase of the intestine of Tilapia nilotica was purified by ammonium sulfate precipitation, followed by PAPTG-Sepharose 4B affinity chromatography, ethylenediamineetetraacetic acid ion-exchange chromatography, polyexchanger PBE 94 chromatofocusing, and Sephadex G-100 gel filtration. β-galactosidase was found to be a single band when examined by poly-acrylamide gel electrophoresis. The purifications of β-galactosidase were 27-fold from the crude extract. β-galactosidase showed optimum activity at pH 5.0 at 40°C, and was specifically found to be able to hydrolyze p -nitrophenyl β-galactopyranoside. It degrades galactan and agarose, and produces galactose. β-galactosidase was strongly inhibited by Hg²⁺ and PCMB. β-galactosidase is considered to be secreted by the upper and middle parts of the intestine and most of the activity was detected in the intestinal juice.     

凡纳滨对虾精氨酸激酶的分离纯化及性质研究

经过CM-纤维素批量层析、Separdex G-100柱层析、DEAE_纤维素柱层析等步骤,从凡纳滨对虾肌肉组织分离得到精氨酸激酶,经SDS-PAGE检测达到电泳纯,分子量约为40kDa.对该酶的性质进行分析结果表明,精氨酸激酶的最适作用温度为55℃,当温度高于65℃时,酶活力显著下降;pH 8时酶活力较高,低浓度的精氨酸对酶活力有促进作用,高浓度时表现抑制作用,而底物类似物精胺和氨基胍则对酶促反应表现出完全的抑制.NaCl,KCl对酶的活力具有促进作用,低浓度(10 mmol·L-1)MgCl2对酶活力表现出激活作用,而CuCl2与MnCl2则表现出完全抑制酶活力,CaCl2与ZnCl2在低浓度时对酶活力无明显影响,但是随着浓度升高,对酶具有抑制作用.     

柔嫩艾美球虫（Eimeria tenella）体外细胞培养

试验比较了玻璃珠层析法，ＤＥＡＥ纤维素层析法，Ｇ３漏斗过滤法３种提纯柔嫩艾美球虫（Ｅｉｍｅｒｉａｔｅｎｅｌｌａ）子孢子的方法，柱高８ｃｍ，直径２００μｍ的玻璃株柱用ｐＨ８．０的Ｒｉｎｇｅｒ’ｓ缓冲液作洗脱提纯子孢子，回收率为６４．１％，子孢子洗脱高峰期集中，且易于无菌操作，柱高５ｃｍ的ＤＥＡＥ纤维素柱用ｐＨ８．０的甘氨酸缓冲液作洗脱液，提纯子孢子回收率为６３．２％，但严格无菌操作困难，Ｇ３漏斗法易     

水力旋流器在多泥沙河流中应用的研究

利用水力旋流器，对水力机械的润滑、轴封用水进行净化，降低泥沙含量，使之达到技术供水标准。本文研究和分析了水力旋流器的结构、各部构件尺寸与净化泥沙性能的关系；指明了水经过旋流器的压力降的确定，使之既满足泥沙沉降的要求，又要保证供水系统的所需的压力值。     

Interaction of bovine immunoglobulin gamma 2 (IgG2) with staphylococcal protein A: evidence for IgG2a and IgG2b sub-subclasses in the bovine

The reactivity of bovine IgG with protein A is confusing with respect to which of the bovine IgG class and subclasses are reactive. We have, therefore, re-examined the interaction of bovine immunoglobulins with protein A. The results presented in this paper indicated that at pH 8.0 protein A binds only immunoglobulin of the IgG2 subclass. The bound IgG2 can be readily recovered from an immobilized protein A column at pH 5.0. Furthermore, the antigenic IgG2 eluted demonstrated two charged species which could readily be separated by ion-exchange chromatography. These results indicate that IgG2 in the bovine exists in two sub-subclasses, IgG2a and IgG2b. The two sub-subclasses of IgG2 could be rapidly isolated with a good yield in two-steps namely protein A affinity chromatography followed by ion exchange chromatography.     

A Method of Purification of Bovine Colostrum slgA

sIgA in bovine colostrum was purified by ultrafiltration and enzymolysis methods in this experiment, and the prepared substance was detected by Western Blot. The purity and yield were up to 73.6% and 65.2%, respectively. This convenient technique offers helpful exploration for production of bovine colostrum sIgA     

Isolation and Purification of Bovine Colostrum sIgA and IgG

To further utilize bioactive substance such as bovine colostrum sIgA and IgG,sIgA and IgG were isolated and purified simultaneously by salting out,ultrafiltration and gel chromatography,etc.The analysis of results were showed quantitatively by non-hydrogenized SDS-PAGE,and quanlities of sIgA and IgG were respectively detected by Western Blot.The results showed that the purity and yield of bovine colostrum sIgA were 85.3%and 42.8%,respectively,while the purity and yield of bovine colostrum IgG were respectively 97.2%and 64.4%.This preparative method provides theoretical and experimental foundation for sIgA industrial production.     

绵羊瘤胃微生物胞外蛋白酶的动态及纯化

本实验用装有永久性瘤胃瘘管的成年新疆细毛羯羊3只,对饲喂前后瘤胃蛋白酶活性的变化规律及瘤胃液离心各组分的酶活比例进行测定,并对离心(12000转/min,30min,4℃)后的瘤胃上清液中的微生物胞外蛋白酶进行纯化。结果表明:①瘤胃蛋白酶活性在饲喂后3h左右最高,以后逐渐下降,至饲喂后8h左右恢复到饲喂前的水平。②瘤胃液中以游离形式存在的蛋白酶占40％,其余附着在微生物和饲料颗粒上。③用硫酸铵对上清液中的微生物胞外蛋白酶进行浓缩,依次经SephadexG—150、DEAE—Sephadex A—50和FPLC(Mono Q HR5/5柱)进行纯化后,得到电泳纯度的胞外蛋白酶品,其活力从1.77个单位/mg蛋白提高到295个单位/mg蛋白,提高了166.7倍,回收率为4.7％。SDS—PAGE测得纯化蛋白酶的分子量为38KD。     

提取纯化过程对金针菇FV-19菌株发酵液多糖相对分子质量的影响

金针菇 FV- 19菌株深层发酵醪液 .经离心、沉淀、透析处理得到发酵液多糖样品 ,并用凝胶色谱法测定其相对分子质量 .探讨了提取纯化过程对发酵液多糖相对分子质量的影响以及贮存期对其相对分子质量的影响 .     

一种虫草代谢产物中保肝活性物质的分离和纯化

通过对交织顶孢霉的发酵菌丝粉的生物活性测定,发现其代谢产物可有效降低由肝损伤引起的丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及脂质过氧化物(LPO)的升高,具有良好的保肝活性.用吸附色谱法、凝胶过滤色谱法等方法对该活性成分进行分离和纯化,并用高效液相色谱法、薄层色谱法和化学方法对其纯度进行检验,得到2个纯物质HG-1和HG-2.