Isolation of lactoperoxidase, lactoferrin, α-lactalbumin, β-lactoglobulin B and β-lactoglobulin A from bovine rennet whey using ion exchange chromatography

A mild and rapid method is described for isolating various milk proteins from bovine rennet whey. β-Lactoglobulin from bovine rennet whey was easily adsorbed on and desorbed from a weak anion exchanger, diethylaminoethyl-Toyopearl. However, α-lactalbumin could not be adsorbed onto the resin. α-Lactalbumin and β-lactoglobulin from rennet whey could also be adsorbed and separated using a strong anion exchanger, quaternary aminoethyl-Toyopearl. The rennet whey was passed through a strong cation exchanger, sulphopropyl-Toyopearl, to separate lactoperoxidase and lactoferrin. α-Lactalbumin and β-lactoglobulin were adsorbed onto quaternary aminoethyl-Toyopearl. α-Lactalbumin was eluted using a linear (0–0.15 M) concentration gradient of NaCl in 0.05 M Tris–HCl buffer (pH 8.5). Subsequently, β-lactoglobulin B and β-lactoglobulin A were eluted from the column with 0.05 M Tris–HCl (pH 6.8), using a linear (0.1–0.25 M) concentration gradient of NaCl. The yields were 1260 mg α-lactalbumin, 1290 mg β-lactoglobulin B and 2280 mg β-lactoglobulin A from 1 l rennet whey.     

Expression of Aspergillus niger N5-5 in E. coli and purification and identification of products

Due to the feature of high hydrolysis, tannase is widely used in food, beverage, brewing and other fields. However, high cost in producing natural tannase makes it difficult to apply tannase to industry in a large-scale. Microbial expression systems can be used for preparing numerous amount of enzyme at low cost, so in this paper Aspergillus niger N5-5 was expressed using E. coli system. Specific primers were designed based on the Aspergillus niger N5-5 sequence N3 (GenBank, No.: KP677552), and tannase gene tan was promoted to carry 6 His tag and enzyme cutting site which contains NdeI/HindIII using PCR amplification. Then, tannase gene tan was connected to expression vector by NdeI/HindIII enzyme cutting. In this way, recombinant expression vector tan-pET43.1a was formed. Then, the expression vector pET43.1a by NdeI/HindIII enzyme cutting was transformed into E. coli BL21 (DE3) to induce expression of Aspergillus niger N5-5. When the induced fungi were disrupted by the ultrasonic wave, the crude enzyme was extracted and purified by using the IMAC, and then the activity of the crude enzyme and pure enzyme was determined. According to the results of determination of the tannase activity, the tannase activity of the crude enzyme was greatly improved after the crude enzyme was purified, and the specific activity of the pure enzyme was about 8 times of that of the crude enzyme. The results of SDS-PAGE of the pure enzyme showed that the molecular mass of the pure enzyme was about 65 kDa/64–65 kDa, which was consistent with the expected result (64.2 kDa), It can be concluded that the crude enzyme solution was purified successfully. The results of pure enzyme’s protein identification by Western Blotting showed that clear protein bands pro-3 were observed. Molecular mass of clear protein bands pro-3 was about 65 kDa, which was in line with the expected results (64.2 kDa). It can be seen that the aforementioned expression protein could be specifically combined with His tag. It proved expression protein to be a recombinant fusion protein with 6 His tag.     

Morphological variation in diadromous and landlocked populations of the spotted galaxias,Galaxias truttaceus,in Tasmania,south-eastern Australia

Synopsis A comparison of a suite of morphometric measurements and meristic counts of individuals of two landlocked lacustrine and two diadromous riverine populations of Galaxias truttaceus was carried out utilising both univariate and canonical variate analyses. Lacustrine fish had fewer dorsal and anal fin rays than did riverine fish. Differences were not as clear for gill rakers and vertebrae. Comparisons of serial counts were made with two derived lacustrine species, G. auratus and G. tanycephalus, also from Tasmania. Lacustrine G. truttaceus varied in the same direction as the derived species, relative to riverine G. truttaceus. From an analysis of 12 body measurements, the first canonical variate clearly separated lacustrine fish from riverine fish largely based on measurements associated with fins (pre-anal fin length, length of anal base, pre-dorsal fin length, maximum length of dorsal fin and inter-orbital width). An overall value for the correct classification of fish into groups based on locality was 84%. The percentage of fish classified into the wrong habitat (lake or stream) was much less than the percentage classified between localities within habitats. Overall morphological variation was greater between than within habitats. It is suggested that the differences in water movement and food type may in part account for the differences shown and that selective pressures peculiar to the lacustrine environment may be causing the lake populations to diverge from the riverine populations.     

中药马蓝叶及其混乱品的比较鉴别

马蓝叶是加工传统中药青黛的主要原料。为了确保药材质量,作者对中药马蓝叶及其混乱品种球花马蓝、疏花马蓝、广西马蓝的原植物、药材性状、叶组织显微特征和薄层层析鉴别等进行了研究。指出其混乱品种虽与马蓝叶是同属植物,但不含马蓝叶的有效成分靛玉红,应仔细鉴别,不宜混用。     

Structure of equine corticotropin releasing factor

A 41 amino acid peptide, probably identical in structure to human corticotropin releasing factor, was isolated from 70 equine hypothalami by methanol extraction, immunoaffinity chromatography and single step of reverse phase HPLC. The amino acid sequence was determined by gas phase sequence analysis. Probable carboxyl terminal amidation was demonstrated by similar retention times for equine and human corticotropin releasing factor on reverse phase HPLC at pH 8. The likely structure of equine corticotropin releasing factor is: Ser-Glu-Glu-Pro-Pro- Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-Glu-Met-Ala-Arg-Ala-Glu-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn- Arg-Lys-Leu-Met-Glu-Ile-Ile-NH₂. The purified peptide is equipotent with human corticotropin releasing factor in an in vitro bioassay and in a human plasma binding protein assay.     

猪垂体总mRNA的提取、鉴定及其cDNA的合成

 用改进的LiCl沉淀法和寡聚(dT)-纤维素亲和层析法由猪垂体制得总mRNA。在兔网织红细胞无细胞翻译体系中进行体外翻译的结果表明,制得的总mRNA具有一定的翻译活力。翻译产物与兔抗猪生长激素抗血清发生免疫沉淀,沉淀物占总翻译产物的10％左右。SDS聚丙烯酰胺凝胶电泳的结果表明翻译产物有一条很深的带,分子量约为24,000道尔顿,与猪前生长激素的分子量相近。以制备的mRNA为模板反转录合成了双链cDNA。第一链的合成产率为10—35％,第二链的合成产率为84—115％。cDNA的平均分子长度为825bp。     

酵母鉴定程序在酵母分类鉴定中的应用

本文介绍了Barnett等1985年编制并由剑桥大学发行的计算机软件《酵母鉴定程序》,以及如何使用该程序在IBM PC DOS操作系统上进行酵母菌的分类鉴定。我们使用该程序对从云南鸡足山和紫金山两地森林土壤中分离到的82株酵母菌进行了分类鉴定,其中:鉴定到种的有47株,占总株数的57.3％;鉴定到属但未能直接定到种的有21株,占总株数的25.6％;暂未定名的有14株,占总株数的17.1％。     

从我国成人手足口病患者疱液中首次分离出肠道病毒71型

本文报道了从我国手足口病(HFMD)患者疱液中肠道病毒71(E71)型H株的分离和鉴定。该株病毒可在原代人胚肺(HEL)细胞、MA104细胞、BSC细胞中生长繁殖,导致典型的肠道病毒CPE出现。将H株接种乳鼠后出现肢体麻痹、第6天开始死亡。电镜下可见感染H株的BSC细胞胞浆中出现大量的结晶状排列的成熟病毒颗粒,直径约25nm。患者双份血清中有对H株4倍增高的中和抗体存在。采用100抗体单位的抗Cox A5、7、9、16、E70、E71抗体和50抗体单位的LBM组合血清A-H以及抗E71BrCr株MeAb P27对H株进行中和试验时,H株可被抗E71血清和MeAb P27所中和。抗E71抗体对H株的最低有效中和作用为1.6抗体单位,MeAb P27对H株的有效中和作用是64抗体单位。其它血清则无此中和作用。然而,在鉴定过程中发现,高滴度的抗Cox A16抗体(200抗体单位以上)也显出有中和H株的作用,提示我们所分离的H株含有与Cox A16的型间共同抗原。