The purification and partial characterization of carp,Cyprinus carpio,vitellogenin

A procedure is described for the isolation of intact vitellogenin (c-VTG) from the carp, Cyprinus carpio. VTG was induced in juvenile females using oestradiol-17β and purified from the plasma using a combination of gel-filtration chromatography on Sepharose 6B and ion exchange chromatography on DEAE-cellulose. Purification procedures were conducted at low temperatures (below 9°C) in the presence of the proteolytic enzyme inhibitor aprotinin to prevent degradation. Intact c-VTG had an apparent molecular mass of 390,000 Daltons, but when extracted from plasma in the absence of aprotinin it underwent proteolysis into at least 2 protein fragments (apparent molecular masses of 230,000 and 96,000 Daltons), showing an instability of the native dimer. An amino acid analysis of c-VTG showed that its composition was almost identical to goldfish VTG, a species closely allied to the true carps and also similar to other oviparous vertebrate VTGs. Collectively, these data indicate that using these purification procedures VTG from carp, and probably other teleost species, can be isolated in an intact, highly purified form.     

Purification of chinook salmon (Oncorhynchus tshawytscha) GH for receptor study

A method for the purification of chinook Salmon (Oncorhynchus tshawytscha) GH, which retains its biological activity, is described. The biological activity was investigated with an established radioreceptor assay using liver membranes from pregnant rabbits and bovine GH as standard and labelled hormone. The enrichment of the preparation was checked with electrophoresis (SDS-PAGE). Extraction and further steps were carried out using low molarity alkaline buffer (pH 8–10, M = 100 mM). Three chromatography steps were performed (Concanavalin-A sepharose, Bio-gel P60, DEAE). Ion exchange chromatography was performed under isocratic conditions (using a 50 cm column). Two isoforms (sGH1 and sGH2) were isolated. The purification yield is 0.7% compared to lyophilized pituitaries. The molecule is homogeneous in SDS-PAGE. Contamination by prolactin, gonadotrophin and corticotrophin is negligible (< 0.5%). It could be demonstrated that the biological activity of the preparation is maintained since this preparation stimulates the growth of juvenile trout (Salmo gairdneri) and binds specifically (35%) to trout liver membranes.     

Protein kinase C in the spleen of the turbot (Scophthalmus maximus L.)

PKC activity was detected in spleen extracts from the turbot, Scophthalmus maximus, a teleost flatfish that is farmed commercially in several countries, in assays with the substrate EGF- R651–658 as phosphate acceptor. The activity was purified about 700-fold by a three-step chromatographic procedure (DEAE-cellulose, phenyl-Sepharose and threonine-Sepharose). Maximal activity was obtained in the presence of the typical PKC cofactors Ca2+ (0.1 mM) PtdS (20 g ml–1) and either DAG (2 g ml–1) or PMA (2 g ml–1). Activity was dose-dependently inhibited by H7 and by the PKC-specific inhibitors PKC19–36 and N-myristoylated PKC19–31. The rate of phosphorylation was highest with the PKC-specific substrate MARCKS161–175. In immunoblotting, MC5 (a mouse monoclonal antibody raised against bovine PKC) recognized bands of 80 and 100 kDa. Immunoblotting with antibodies raised against mouse PKC isozymes (, , , , , , and ) indicated the presence of all these isozymes in turbot spleen.     

鲢肠道淀粉酶的分离纯化及部分性质研究

以鲢(Hypophthalmichthys molitrix)加工下脚料为原料,分离提纯其中的淀粉酶并对其部分性质进行了研究。结果表明经多级分离,得到了一种相对分子质量约2.15×104的淀粉酶,命名为SCIG1。SCIG1是鲢肠道淀粉酶的一种,酶活性的最适温度为30℃,最适pH5.5。     

以PLA、PHBV为碳源的生物絮团技术在海水养殖中的应用

本研究以聚乳酸(PLA)和3-羟基丁酸-co-3-羟基戊酸共聚(PHBV)作为外加碳源,探究在模拟海水生物絮团养殖中,高、中、低盐度下PLA与PHBV碳源释放规律以及对海水生物絮团养殖中水质、微生物多样性及其群落结构的影响.结果显示:PHBV碳源要优于PLA碳源,中盐度更有利于各种营养盐的转化,氨氮质量浓度最终保持在3...     

贝类净化中试生产工艺技术

本文介绍用紫外线法消毒的海水对文蛤、花蛤、缢蛏进行净化的中试生产工艺技术及其净化效果。通过对该三种净化贝净化前后的各种安全指标以及其净化过程中大肠菌群数和砂份含量随时间的变化进行检测,认为原料符合SC/T 3013-2002规范的文蛤、缢蛏、花蛤经24小时的净化其大肠菌群都能达到DB35/575-2004净化海水贝类标准的要求(≤300MPN/100g贝肉),同时大肠群菌数较低的贝类可以缩短净化时间;根据其含砂量的不同净化时间可控制在8~16小时;要通过净化来大量降低贝类的重金属、农药及贝毒的含量似乎是不可能的;根据贝类品种的不同,其净化过程的死亡率在1.1~3.5%,产品得率90~98%;其净化产品在8℃~10℃贮存与流通可贮藏7天。     

橡胶树肌动蛋白解聚因子原核表达及纯化

本研究根据HbADF序列设计引物扩增基因的编码区,并将其插入到原核表达载体pET28a上,成功构建重组质粒pET28a-HbADF。将重组质粒转化大肠杆菌BL21(DE3)宿主菌,经1 mmol/L IPTG诱导,获得相对分子量为20 ku的融合蛋白。表达蛋白以可溶和包涵体两种形式存在,通过亲和层析方法纯化可溶蛋白并获得HbADF融合蛋白,用抗HIS标签的单抗对纯化蛋白进行了Western blot鉴定。该结果为进一步研究HbADF蛋白特性及功能奠定基础。     

高速逆流色谱法制备分离库拉索芦荟中的2’-对香豆酰芦荟宁

采用高速逆流色谱法分离纯化库拉索芦荟中的2’-对香豆酰芦荟宁。以正己烷-乙酸乙酯-丙酮-水(1∶5∶1∶5,V/V/V/V)为溶剂体系,上相为固定相,下相为流动相,线圈转速为860 r/min,流速为2 mL/min,紫外检测波长为300 nm,一次进样85 min内从487.5 mg库拉索芦荟丙酮提取物中分离得到19.5 mg单体组分,经高效液相色谱分析,测定纯度为95.6%(峰面积百分比),通过质谱、红外、紫外和核磁技术进行结构鉴定,确定组分为2’-对香豆酰芦荟宁。该法具有操作简便、速度快、样品制备量大、节省溶剂及回收率高等优点,可为芦荟宁类化合物的分离提供高效、稳定及可靠的方法。     

一种EGCG单体纯化新工艺

以粗酯型儿茶素为原料,研究采用大孔吸附树脂和结晶纯化儿茶素单体EGCG的制备工艺。通过比较4种大孔吸附树脂对EGCG的吸附和解吸能力,筛选出最优树脂AB-8,并考察乙醇梯度洗脱等特性,再经过结晶析出EGCG晶体。结果表明:AB-8大孔吸附树脂对EGCG具有较好的吸附选择性,通过10%乙醇洗脱,EGCG产品纯度从54.41%提高到了92.65%,得率达93.38%,结晶后得纯度达98%以上的EGCG单体。     

打叶复烤风选除杂烟叶质量分析

该文从外观质量、叶片结构、常规化学和感官质量4个方面对风选除杂烟叶进行了分析。结果表明：与正常B2F烟叶相比,风选除杂烟叶色泽暗淡,轻质,油分少,身份薄;大片率、大中片率、中片率、叶中含梗率相对较低,而小片率、碎片率正好相反;总糖、还原糖、烟碱含量相对较低,而总氮、钾含量正好相反,氯含量两者接近;香气量不足,劲头、浓度小,杂气、刺激性大,余味较苦涩,燃烧性好,呈灰白色,总体使用价值较差。