蚯蚓纤溶酶的分离纯化研究

目的从赤子爱胜蚓(Eisenia Foelide)中分离纯化出一种相对分子质量(Mr)较小的纤维蛋白溶解组分———蚯蚓纤溶酶(EFE),为其注射剂的研制开发打下基础。方法采用盐析、透析、凝胶过滤色谱、离子交换色谱等方法分离纯化EFE,用SDS-PAGE对纯化的活性组分进行Mr测定,用酶谱分析方法初步探讨蚯蚓中存在的其它纤溶酶活性组分。结果分离纯化得到了EFE单一组分,经SDS-PAGE EFE呈单一条带,Mr约为25 000。经酶谱分析发现纤溶酶活性组分Mr分布在25 000~50 000之间。结论反复使用色谱技术可分离纯化EFE,并得到单一组分。     

核酸酶P1的分离纯化及部分酶学性质研究

桔青霉发酵液经离心、超滤、硫酸铵盐析、透析和DE-52型树脂柱色谱分离,纯化得到核酸酶P1.比活为28490u/mg,纯化10.5倍,收率为45%.SDS-PAGE电泳显示为单一条带.酶学性质研究结果表明,以酵母RNA为底物时的Km为3.36mmol/L,最适作用温度70℃,最适pH 5.4.     

降纤酶的聚乙二醇修饰及产物的纯化研究

研究了降纤酶的聚乙二醇修饰及纯化方法。采用正交法考察影响修饰的4个因素，确定了最佳条件。利用分子筛柱色谱得到了较高酶活力保留的修饰产物，SDS-PAGE检测为单一条带。     

马粪海胆生殖腺中多糖的纯化方法及其抗肿瘤作用研究

目的 优化马粪海胆生殖腺中多糖的纯化方法,并评价其体外对肿瘤细胞的生长抑制作用.方法 马粪海胆生殖腺经水提取、木瓜蛋白酶结合Sevage法除蛋白、透析后醇沉、干燥制得马粪海胆总多糖,总多糖应用Sephacryl S-300凝胶柱色谱分离纯化;采用苯酚-硫酸法测定多糖含量;通过噻唑蓝(MTT)法测试多糖的体外抗肿瘤作用.结果 马粪海胆生殖腺的水提液应用木瓜蛋白酶结合Sevage法除蛋白效果最佳,木瓜蛋白酶最佳用量为100∶6(海胆脱脂粉与酶质量比),制得总多糖的纯度达90%;总多糖及分离得到的2个多糖体外对人肝癌Bel7402细胞表现出生长抑制作用.结论 该纯化方法步骤简单、易操作,所得多糖理化性质好、易溶于水,适用于马粪海胆生殖腺多糖的纯化.     

尖吻蝮蛇毒中一种新类凝血酶的分离纯化

用阴离子交换色谱和凝胶过滤色谱技术,从尖吻蝮蛇毒中纯化得到一个具有凝血活性的组分.经Superdex 75 HR10/30预装柱检测,纯度达99.91%.SDS-PAGE显示为单一条带,还原、非还原条件下分子量分别为59.25k、52.58k.该组分的凝血酶比活为41.5u/mg,精氨酸酯酶比活为14.29u/mg,但不激活血浆凝血因子ⅩⅢ.EDTA不能抑制其凝血活性,而苯甲磺酰氟则产生不可逆抑制作用.结果表明该酶是一种新尖吻蝮蛇毒类凝血酶.     

西兰花种子中分离纯化萝卜硫素及残留溶剂的方法学验证

目的 介绍了一种从西兰花种子中分离纯化天然萝卜硫素高效、低成本方法。方法 该方法包括种子自溶后的溶剂萃取、柱色谱分离和半制备高效液相色谱纯化。经MS、¹H-NMR、¹³C-NMR谱对纯化后的化合物进行了表征。结果 与传统的液-液萃取法相比，柱色谱分离法从西兰花种子中提取萝卜硫素的收率更高为(69.02±1.1)%。以SinoChrom ODS-BP柱为半制备高效液相色谱柱，以甲醇-水(体积比1∶4)为流动相，最大装载量小于60 mg,流量应控制在9 mL·min^-1,可以获得纯度为95%的萝卜硫素。经HPLC、MS、¹H-NMR、¹³C-NMR检测，结果表明，所得样品为目标化合物，且无有机溶剂残留。结论 建立了一种分离纯化西兰花种子中萝卜硫素的方法。本方法经济、高效，有实用价值。     

不同来源的硫酸乙酰肝素的制备及理化性质研究

目的从不同来源制备硫酸乙酰肝素 (HS) ,并测定其理化性质。方法利用酶解法提取分离糖胺聚糖混合物 ,采用乙醇分级沉淀和离子交换柱色谱、凝胶过滤柱色谱等方法将HS纯化 ,醋酸纤维素薄膜电泳 (CAME)测定其纯度。结果所得各种来源的HS在CAME电泳显示一条带 ,且理化性质、生物活性和红外光谱各不相同。结论不同来源的HS具有不同的结构和性质     

牛肝β-D-葡萄糖醛酸苷酶的分离纯化和性质研究

采用自溶、35 %硫酸铵分部沉淀、CM 纤维素柱层析、羟基磷灰石柱层析由动物肝脏中分离纯化β D 葡 萄糖醛酸苷酶 ,获得比活 2 6 70 0U/mg的纯酶 ,活力回收率为 4 2 .8% ,纯化的酶经SDS PAGE呈单一条带。该酶以 β D 酚酞葡萄糖醛酸苷为底物的Km值为 2 .0× 10 -4mol/L ,以对硝基苯酚葡萄糖醛酸苷为底物的Km值为 2 .5× 10 -4mol/L。在 pH值 5～ 6的范围内 ,该酶的活力较高 ,最适 pH值为 5 .5 ,在pH值 5 .5时稳定性最好。在 30～ 5 0℃范围内酶活力均较稳定     

通过亲和色谱从地衣芽孢杆菌中分离α-淀粉酶

建立了使用新型洗脱液的亲和色增方法从地衣芽孢杆菌中分离纯化α-淀粉酶。结合到淀粉、淀粉-硅藻土、淀粉-琼脂糖柱上的α-淀粉酶可用2％（w／v）白糊精快速洗脱。淀粉柱结合α-淀粉酶的能力为380gmol／g。纯化后的α-淀粉酶在SDS-PAGE凝胶电泳中显示58k的单一条带。用免疫扩散、免疫电泳确定纯化后酶的特征，并用单向辐射状免疫扩散和western免疫杂交研究了该酶在不同时间的合成情况。     

纳豆激酶的分离纯化及酶学性质研究

目的研究纳豆激酶分离纯化工艺及酶学性质。方法纳豆激酶发酵液的粗提物经Superdex 75凝胶色谱和聚丙烯酰胺凝胶电泳(PAGE)分离纯化,采用TAME法测定酶的活性,通过SDS-PAGE对纯化结果进行了检验。结果SDS-PAGE中显示单一色带,相对分子质量28000,以TAME为底物时纳豆激酶的米氏常数(Km)为35.47mmol/L,最适宜的温度37℃,最适宜pH为8.6。结论该分离纯化方法可以得到较纯的纳豆激酶。     

Pharmacological study of papain from the papaya plant cultivated in Uzbekistan

It was experimentally established that papain from papaya cultivated in Uzbekistan possesses a pronounced proteolytic activity: 0.1, 0.5, and 1% papain solutions decreased the weight of burn crust in vitro and accelerated experimental burn healing in vivo. Under clinical conditions, papain produced therapeutic effect in patients with inflammatory disorders in genitals, intestine, liver, and eye. The pharmacological effects of papain produced from Uzbek papaya are identical to those of the commercial product from Merck.     

Therapeutic application of Carica papaya leaf extract in the management of human diseases

IntroductionPapaya (Carica papaya Linn.) belongs to the family Caricaceae and is well known for its therapeutic and nutritional properties all over the world. The different parts of the papaya plant have been used since ancient times for its therapeutic applications. Herein, we aimed to review the anticancer, anti-inflammatory, antidiabetic and antiviral activities of papaya leaf.MethodsAll information presented in this review article regarding the therapeutic application of Carica papaya leaf extract has been acquired by approaching various electronic databases, including Scopus, Google scholar, Web of science, and PubMed. The keywords Carica papaya, anticancer, anti-inflammatory, immunomodulatory, and phytochemicals were explored until December 2019.ResultsThe papaya plant, including fruit, leaf, seed, bark, latex, and their ingredients play a major role in the management of disease progression. Carica papaya leaf contains active components such as alkaloids, glycosides, tannins, saponins, and flavonoids, which are responsible for its medicinal activity. Additionally, the leaf juice of papaya increases the platelet counts in people suffering from dengue fever.ConclusionThe major findings revealed that papaya leaf extract has strong medicinal properties such as antibacterial, antiviral, antitumor, hypoglycaemic and anti-inflammatory activity. Furthermore, clinical trials are needed to explore the medicative potential of papaya leaf. Graphical abstractOpen in a separate windowGraphical abstract showing the medicinal properties of Carica papaya leaf     

Identification and detection method for genetically modified papaya resistant to papaya ringspot virus YK strain

Unauthorized genetically modified (GM) papaya (Carica papaya LINNAEUS) was detected in a commercially processed product, which included papaya as a major ingredient, in Japan. We identified the transgenic vector construct generated based on resistance to infection with the papaya ringspot virus (PRSV) YK strain. A specific detection method to qualitatively monitor papaya products for contamination with the GM papaya was developed using the real-time polymerase chain reaction.     

Effect of dried fruits of Carica papaya Linn on hepatotoxicity

Ethanol and aqueous extracts of Carica papaya has been evaluated for its anti hepatotoxic activity. The ethanol and aqueous extracts of Carica papaya showed remarkable hepatoprotective activity against CCl(4) induced hepatotoxicity. The activity was evaluated by using biochemical parameters such as serum aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase, total bilirubin and gamma glutamate transpeptidase (GGTP). The histopathological changes of liver sample was compared with respect to control.     

Anti-inflammatory activities of ethanolic extract of <Emphasis Type="Italic">Carica papaya</Emphasis> leaves

The anti-inflammatory activity of an ethanolic extract of Carica papaya leaves was investigated in rats using carrageenan induced paw oedema, cotton pellet granuloma and formaldehyde induced arthritis models. Experimental animals received 25-200 mg/Kg (orally) of the extracts or saline (control group) and the reference group received 5 mg/ Kg of indomethacin. The ulcerogenic activity of the extract was also investigated. The results show that the extracts significantly (p <0.05) reduced paw oedema in the carrageenan test. Likewise the extract produced significant reduction in the amount of granuloma formed from 0.58 +/-0.07 to 0.22 +/-0.03 g. In the formaldehyde arthritis model, the extracts significantly reduced the persistent oedema from the 4th day to the 10th day of the investigation. The extracts also produced slight mucosal irritation at high doses. The study establishes the anti-inflammatory activity of Carica papaya leaves.     

Effect of ether- and water-soluble fractions of Carica papaya ethanol extract in experimentally induced hyperlipidemia in rats

Context: The papaya is the fruit of the plant Carica papaya L. (Caricaceae) used in India. Fruit and latex are both rich in an enzyme called papain. It is used as a folk remedy for contraception and abortion.

Objective: The present study explored the anti-hyperlipidemic effect of the ether- and water-soluble fractions of C. papaya ethanol extract in olive oil-induced hyperlipidemic rats. The study also involved chromatographic studies of extract and fractions.

Materials and methods: Flash chromatography was done for the most active fraction. The extract and fractions were administered orally at doses of 200 and 400?mg/kg body weight in rats. Olive oil (5?mL/kg oral dose) was administered 30?min after treatment. Blood was collected and centrifuged at 3000?rpm for 15–20?min and subjected to biochemical analysis.

Result: The study dose-dependently inhibited the total cholesterol (TC), triglycerides (TG), low-density lipoproteins (LDL) level, and significantly increased high-density lipoprotein (HDL) level. Phytochemical screening revealed the presence of fats in the ether fraction, whereas the water fraction revealed the presence of tannins, alkaloids, glycosides. UV λ_max was found to be 217?nm with a melting point of 41°C for the isolated component.

Discussion and conclusion: The anti-hyperlipidemic effect was evaluated in olive oil-loaded rats. Acute treatment caused stimulatory effect on HDL level and inhibition in TC and TG elevation induced by olive oil. The extract and water fraction showed protective action by increasing the HDL cholesterol level.     

云南峨山番木瓜中木瓜蛋白酶的初步纯化及活性测定

应用FPLC系统对采自云南峨山县的番木瓜乳汁中的木瓜蛋白酶进行了初步纯化,同时,用基梅尔-史密斯法测定了番木瓜乳汁中木瓜蛋白酶的活性。结果表明:不同瓜形之间的木瓜蛋白酶活性大小没有明显差异,但与瓜的成熟度有关。     

The invitro assessment of antibacterial effect of papaya seed extract against bacterial pathogens isolated from urine, wound and stool

BACKGROUND: Carica papaya family Caricaceacae is one of the herbal remedies, which has recently become a subject of research focus. It is used in traditional medicine for variety of purposes in treating infectious and noninfectious diseases. OBJECTIVE: The objective of this study was to assess the antibacterial effect of papaya seed extract against bacterial pathogens isolated from wound, urine and stool. METHODS: This analytical experimental study was conducted in Jimma University, School of Medical Laboratory Technology, Microbiology laboratory between February to March 2005. The antibacterial activity of methanol extract of papaya seed was investigated against specific pathogenic bacteria isolated from wound, urine and stool by an agar dilution technique and the crude preparation was assessed by an agar diffusion technique. The growth or inhibition of control strains of Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa as well as the clinical isolates of these bacteria were determined in growth media. RESULTS: Results obtained in this study indicate that the minimum inhibitory concentration of papaya seed extract for 50% of the test bacteria was 18.38mg/ml and for S. typhi the MIC was at 11.8 mg/ml of extract. However, the growth inhibitory effect of papaya seed extract was not observed for P. aeruginosa up to 26.25 mg /ml of extract. Even though, the minimum bactericidal concentration is higher than the minimum inhibitory concentration of papaya seed extract (13.13 mg/ml, 11.8 mg/ml respectively) against S. typhi control and clinical isolates, the minimum bactericidal concentration for 50% of the tested bacteria was found to be similar with the minimum inhibitory concentration of the test bacteria, CONCLUSION: Papaya seed could be used as an effective antibacterial agent for the tested organisms. Nevertheless, preclinical studies including invivo animal models and clinical trial on the effect of the seed are essential before advocating large-scale therapy.     

Effect of papaya (Carica papaya linn) on pregnancy and estrous cycle in albino rats of Wistar strain

The antifertility effects of Carica papaya were investigated by feeding adult cycling and pregnant rats with different components of its fruits. No attempt was made to force-feed the animals. The results indicate that unripe fruits of papaya interrupt estrous cycle and induce abortions. The abortifacient property seems to decrease as the fruit becomes stale or ripe. Exogenous progesterone counteracts partially the adverse effects on pregnancy and the surviving foeti are without any distinct malformations.     

木瓜凝乳酶基因克隆及毕赤酵母分泌型重组表达载体构建

目的扩增木瓜凝乳酶基因 ,构建重组表达载体。方法提取木瓜组织RNA ,采用RT PCR方法扩增木瓜凝乳酶基因 ,测序并利用BLAST软件进行序列分析。利用基因重组方法构建重组表达载体。结果扩增得到木瓜凝乳酶基因。结论构建了可在毕赤酵母表达系统中表达天然木瓜凝乳酶蛋白的重组表达载体。