Molecular Weight

分子量
  • 文章类型: Journal Article
    花粉,除了过敏原,包含小于3kDa的低分子量组分(LMC)。新出现的证据表明LMC在过敏性免疫应答中的相关性。然而,桦树花粉(BP)来源的LMC与上皮细胞的相互作用尚未得到广泛研究。我们调查了暴露于BPLMC诱导的上皮屏障修饰,使用人支气管上皮细胞系16HBE14-。将上皮细胞单层顶部暴露于主要的BP过敏原Betv1,水性BP提取物或BP衍生的LMC。通过定期测量跨上皮电阻并使用xCELLigence实时细胞分析系统来监测处理后的屏障完整性。使用多重免疫测定法测量治疗后24小时细胞因子的极化释放。暴露于BPLMC后,上皮屏障完整性显着增强。此外,BPLMC诱导木瓜蛋白酶介导的上皮屏障损伤修复。暴露于BPLMC后,CCL5和TNF-α的顶端释放显着降低,而基底外侧释放的IL-6显著增加。总之,我们的研究结果表明,BP衍生的LMC改变了支气管上皮细胞的物理和免疫特性,从而调节了气道上皮屏障反应。
    Pollen, in addition to allergens, comprise low molecular weight components (LMC) smaller than 3 kDa. Emerging evidence indicates the relevance of LMC in allergic immune responses. However, the interaction of birch pollen (BP)-derived LMC and epithelial cells has not been extensively studied. We investigated epithelial barrier modifications induced by exposure to BP LMC, using the human bronchial epithelial cell line 16HBE14o-. Epithelial cell monolayers were apically exposed to the major BP allergen Bet v 1, aqueous BP extract or BP-derived LMC. Barrier integrity after the treatments was monitored by measuring transepithelial electrical resistance at regular intervals and by using the xCELLigence Real-Time Cell Analysis system. The polarized release of cytokines 24 h following treatment was measured using a multiplex immunoassay. Epithelial barrier integrity was significantly enhanced upon exposure to BP LMC. Moreover, BP LMC induced the repair of papain-mediated epithelial barrier damage. The apical release of CCL5 and TNF-α was significantly reduced after exposure to BP LMC, while the basolateral release of IL-6 significantly increased. In conclusion, the results of our study demonstrate that BP-derived LMC modify the physical and immunological properties of bronchial epithelial cells and thus regulate airway epithelial barrier responses.
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  • 文章类型: Journal Article
    三角phaeodactylum是通过其快速生长的能力来识别的,繁殖,和体外培养,以及一系列高价值活性化合物的存在,包括蛋白质,具有潜在的食品应用。这项研究的目的是研究pH值偏移处理(pH值分别为3、5、7、9和11)对三角斑藻蛋白(PTP)的结构和功能特性的影响。PTP的分子量主要分布在以下范围内:低于5kDa,5-100kDa,超过100kDa。与酸性环境相比,PTP在碱性环境中表现出更高的溶解度和更大的游离巯基含量。此外,PTP在碱性环境中具有较小的粒径和较高的热稳定性。PTP表现出优异的发泡能力(135%),乳化活性指数(3.72m2/g),和乳液稳定性指数(137.71min)在碱性环境中。该研究结果为PTP在食品工业中的未来发展和应用提供了基础。
    Phaeodactylum tricornutum is identified by its capacity for rapid growth, reproduction, and in vitro cultivation, as well as the presence of a range of high-value active compounds, including proteins, with potential food applications. The objective of this study was to investigate the effects of pH shift treatments (pH of 3, 5, 7, 9, and 11) on the structural and functional properties of the Phaeodactylum tricornutum protein (PTP). The molecular weight of the PTP was predominantly distributed within the following ranges: below 5 kDa, 5-100 kDa, and above 100 kDa. Compared to the acidic environment, the PTP demonstrated higher solubility and greater free sulfhydryl group content in the alkaline environment. Additionally, PTP had a smaller particle size and higher thermal stability in alkaline environments. The PTP exhibited superior foaming ability (135%), emulsification activity index (3.72 m2/g), and emulsion stability index (137.71 min) in alkaline environments. The results of this investigation provide a foundation for the future development and application of the PTP in the food industry.
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  • 文章类型: Journal Article
    背景:细菌基因组通常编码类似于噬菌体衣壳(封装蛋白)和噬菌体尾的结构,其可以自发诱导或使用基因毒性化合物如丝裂霉素C。这些高分子量(HMW)推定的抗细菌蛋白(ABP)在自然环境下用于对抗竞争性菌株。以前,尚不清楚这些HMW推定的ABP是否源自昆虫致病性革兰氏阳性,孢子形成细菌侧孢短芽孢杆菌(Bl)分离株(1821L,1951)是在生长过程中自发诱导的,对它们自己的生存产生不利影响。此外,以前没有进行任何工作来确定它们的生化特性。
    结果:使用聚乙二醇沉淀的软琼脂覆盖法,从Bl1951的沉淀裂解物中发现了窄谱的生物活性。丝裂霉素C诱导的滤液的电子显微照片显示类似于噬菌体衣壳和收缩尾的结构。在Bl1821L和Bl1951生长期间提取的无细胞上清液(CFS)的生物活性测定表明这些具有自杀活性的HMW推定的ABP的自发诱导。自发诱导的推定ABP的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示,在细菌生长过程中的所有时间间隔中,除了最初的几个小时外,都出现了〜30kDa和〜48kDa的强度不同的条带。统计上,自发诱导的HMW推定的Bl1951ABP在液体中生长18小时后,其生产菌株的活细胞数量显着减少。此外,注意到这一特定时间段的CFS的pH值和显着的生物活性发生了显着变化。生物化学,例如,来源于Bl1821L或Bl1951的过滤的上清液在宽范围的pH和温度下保持生物活性。
    结论:本研究报告了在生长过程中Bl1821L和Bl1951分离株的HMW推定的ABP(细菌素)的自发诱导,其具有潜在的自杀活性,这在作为潜在的生物农药的生产过程中是至关重要的。发现了Bl1951沉淀物的推定抗菌活性的窄谱。Bl1821L和Bl1951的HMW推定ABPs在宽范围的pH和温度下的稳定性可用于将这种有用细菌的潜力扩大到超出杀虫值。
    BACKGROUND: Bacterial genomes often encode structures similar to phage capsids (encapsulins) and phage tails which can be induced spontaneously or using genotoxic compounds such as mitomycin C. These high molecular-weight (HMW) putative antibacterial proteins (ABPs) are used against the competitive strains under natural environment. Previously, it was unknown whether these HMW putative ABPs originating from the insect pathogenic Gram-positive, spore-forming bacterium Brevibacillus laterosporus (Bl) isolates (1821L, 1951) are spontaneously induced during the growth and pose a detrimental effect on their own survival. Furthermore, no prior work has been undertaken to determine their biochemical characteristics.
    RESULTS: Using a soft agar overlay method with polyethylene glycol precipitation, a narrow spectrum of bioactivity was found from the precipitated lysate of Bl 1951. Electron micrographs of mitomycin C- induced filtrates showed structures similar to phage capsids and contractile tails. Bioactivity assays of cell free supernatants (CFS) extracted during the growth of Bl 1821L and Bl 1951 suggested spontaneous induction of these HMW putative ABPs with an autocidal activity. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of spontaneously induced putative ABPs showed appearance of ~ 30 kDa and ~ 48 kDa bands of varying intensity across all the time intervals during the bacterial growth except in the initial hours. Statistically, spontaneously induced HMW putative ABPs of Bl 1951 exhibited a significant decrease in the number of viable cells of its producer strain after 18 h of growth in liquid. In addition, a significant change in pH and prominent bioactivity of the CFS of this particular time period was noted. Biochemically, the filtered supernatant derived from either Bl 1821L or Bl 1951 maintained bioactivity over a wide range of pH and temperature.
    CONCLUSIONS: This study reports the spontaneous induction of HMW putative ABPs (bacteriocins) of Bl 1821L and Bl 1951 isolates during the course of growth with potential autocidal activity which is critically important during production as a potential biopesticide. A narrow spectrum of putative antibacterial activity of Bl 1951 precipitate was found. The stability of HMW putative ABPs of Bl 1821L and Bl 1951 over a wide range of pH and temperature can be useful in expanding the potential of this useful bacterium beyond the insecticidal value.
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  • 文章类型: Journal Article
    住院的COVID-19患者显示出明显的血栓栓塞发生率和死亡风险增加。在这种情况下,尚不清楚Xa因子抑制剂是否优于依诺肝素。因此,有必要进行直接比较,以评估Xa因子抑制剂与依诺肝素在住院COVID-19患者中的预防效果和安全性.
    MEDLINE,Embase,我们在CochraneCentral数据库中搜索了随机对照试验(RCT)或回顾性研究,这些研究比较了Xa因子抑制剂和依诺肝素在COVID-19住院患者中预防血栓栓塞的有效性或安全性.栓塞发生率,出血发生率,全因死亡率是关注的结局之一.使用Mantel-Haenszel加权随机效应模型计算95%CI的相对风险(RR)。
    分析包括6项随机对照试验和2项包含4048例患者的回顾性研究。荟萃分析表明,与低分子量肝素(LMWH)相比,使用Xa因子抑制剂的患者的栓塞发生率[风险比(RR)0.64(95%,CI0.42,0.98);P=0.04,I2=12%]。根据研究设计类型的亚组分析,在RCTs(RR:0.62;95%CI:0.33-1.17;P=0.14)或观察性研究(RR:0.53;95%CI:0.23-1.26;P=0.15)中,使用Xa因子抑制剂的患者与出血发生率无显著相关性[RR0.76(95%CI0.36,1.61);P=0.47,全部原因0.48通过研究设计类型的亚组分析获得了一致的结果。
    因子Xa抑制剂比依诺肝素更有效地预防COVID-19患者的血栓栓塞。有必要进行其他严格的RCT比较因子Xa抑制剂与依诺肝素。
    UNASSIGNED: Hospitalized patients with COVID-19 have shown a significant occurrence of thromboembolism and a heightened risk of death. It remains unclear whether factor Xa inhibitors are superior to enoxaparin in this context. Hence, there is a need for a direct comparison to assess the preventive effects and safety of factor Xa inhibitors versus enoxaparin in hospitalized COVID-19 patients.
    UNASSIGNED: MEDLINE, Embase, and Cochrane Central databases were searched for randomized controlled trials (RCTs) or retrospective studies that compared the effectiveness or safety of factor Xa inhibitors and enoxaparin in preventing thromboembolism in hospitalized patients with COVID-19. Embolic incidence, incidence of bleeding, and all-cause mortality were among the outcomes of interest. Mantel-Haenszel weighted random-effects model was used to calculate relative risks (RRs) with 95 percent CIs.
    UNASSIGNED: The analysis included six RCTs and two retrospective studies containing 4048 patients. Meta-analysis showed a statistically significant reduction among patients on factor Xa inhibitors compared with low-molecular-weight heparin (LMWH) in the embolic incidence [risk ratio (RR) 0.64 (95%, CI 0.42, 0.98); P=0.04, I2=12%]. Upon subgroup analysis by type of study design, no significant reductions were noted in patients on factor Xa inhibitors in RCTs (RR: 0.62; 95% CI: 0.33-1.17; P=0.14) or observational studies (RR: 0.53; 95% CI: 0.23-1.26; P=0.15) when compared with enoxaparin Factor Xa inhibitors were not significantly associated with incidence of bleeding [RR 0.76 (95% CI 0.36, 1.61); P=0.47, I2=0%] or all-cause mortality (RR: 0.81; 95% CI: 0.48-1.36; P=0.43). Consistent results were obtained upon subgroup analysis by the type of study design.
    UNASSIGNED: Factor Xa inhibitors are more effective than enoxaparin in preventing thromboembolism among patients with COVID-19 who are not acutely ill and are hospitalized. Additional rigorous RCTs comparing factor Xa inhibitors with enoxaparin are warranted.
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  • 文章类型: Journal Article
    低分子量有机酸(LMWOAs)是土壤的重要组分,在调控重金属地球化学行为中起着关键作用。生物炭(BC)是一种常用的改良剂,可以改变土壤中的LMWOA。这里,草酸(OA)的四种LMWOA,酒石酸(TA),苹果酸(MA),评估了柠檬酸(CA)在使用(S1-BC)或不使用泡桐生物废弃物产生的BC(S1)的污染土壤中改变Cd和SB解吸行为中的作用。结果表明,OA,TA,MA,随着浓度的增加,CA降低了土壤的pH值,和生物炭部分抵消了LMWOAs的pH降低。LMWOAs在低浓度时减少了土壤中Cd的解吸,但在高浓度时增加了Cd的解吸。CA在这方面是最强大的。LMWOAs对Sb解吸有类似的影响,CA是LMWOA中最有效的物种。向土壤中添加BC通过减少Cd解吸但增加Sb从土壤中解吸并增加Cd的分配系数(Kd)值但降低Sb的Kd值来影响Cd和Sb的动力学。这项研究有助于了解在生物炭存在下LMWOAs对Cd和Sb地球化学行为的影响,以及生物炭改良剂增强Sb从污染土壤中解吸的潜在风险。
    Low molecular weight organic acids (LMWOAs) are important soil components and play a key role in regulating the geochemical behavior of heavy metal(loid)s. Biochar (BC) is a commonly used amendment that could change LMWOAs in soil. Here, four LMWOAs of oxalic acid (OA), tartaric acid (TA), malic acid (MA), and citric acid (CA) were evaluated for their roles in changing Cd and SB desorption behavior in contaminated soil with (S1-BC) or without BC (S1) produced from Paulownia biowaste. The results showed that OA, TA, MA, and CA reduced soil pH with rising concentrations, and biochar partially offset the pH reduction by LMWOAs. The LMWOAs reduced Cd desorption from the soil at low concentrations but increased Cd desorption at high concentrations, and CA was the most powerful in this regard. The LMWOAs had a similar effect on Sb desorption, and CA was the most effective species of LMWOAs. Adding BC to the soil affects Cd and Sb dynamics by reducing the Cd desorption but increasing Sb desorption from the soil and increasing the distribution coefficient (Kd) values of Cd but lowering the Kd values of Sb. This study helped understand the effects of LMWOAs on the geochemical behavior of Cd and Sb in the presence of biochar, as well as the potential risks of biochar amendment in enhancing Sb desorption from contaminated soil.
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    文章类型: Journal Article
    人免疫球蛋白产品用于治疗多种疾病,例如由于完全不存在抗体或产生有缺陷的免疫球蛋白而导致的原发性或继发性免疫缺陷和自身免疫性疾病。人免疫球蛋白产品的质量控制对于确保治疗功能和安全性至关重要。这包括检测Fc功能和抗互补活性(ACA),以及使用欧洲药典(Ph。欧尔.)专著0338、0918、2788和1928年。为此,必须使用特定的生物参考制剂(BRP)。Ph的股票。欧尔.人类免疫球蛋白(分子大小)BRP处于低位,因此由欧洲药品质量与医疗保健局(EDQM)进行了一项合作研究,在生物标准化计划的主持下,校准替换批次。18个实验室,包括制造商和官方药物控制实验室,参加了这项研究。评估了三批候选BRP,并与Ph进行了比较。欧尔.人免疫球蛋白(分子大小)BRP3确保连续性。根据研究结果,候选BRP由Ph.欧尔.委员会作为Ph。欧尔.人免疫球蛋白(分子大小)BRP批次4、5和6。
    Human immunoglobulin products are used for the treatment of a number of diseases, such as primary or secondary immunodeficiencies and autoimmune conditions due to the complete absence of antibodies or the production of defective immunoglobulins. Quality control of human immunoglobulin products is essential to ensure therapeutic functionality and safety. This includes testing for Fc function and anticomplementary activity (ACA), as well as verification of appropriate molecular size distribution using size-exclusion chromatography as prescribed in the European Pharmacopoeia (Ph. Eur.) monographs 0338, 0918, 2788 and 1928. To this end, specific biological reference preparations (BRPs) must be used. Stocks of the Ph. Eur. Human immunoglobulin (molecular size) BRP were running low and therefore a collaborative study was run by the European Directorate for the Quality of Medicines & HealthCare (EDQM), under the aegis of the Biological Standardisation Programme, to calibrate replacement batches. Eighteen laboratories, including manufacturers and Official Medicines Control Laboratories, took part in the study. Three batches of candidate BRPs were assessed and compared to Ph. Eur. Human immunoglobulin (molecular size) BRP 3 to ensure continuity. Based on the study results, the candidate BRPs were adopted by the Ph. Eur. Commission as Ph. Eur. Human immunoglobulin (molecular size) BRP batch 4, 5 and 6.
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  • 文章类型: Journal Article
    在这项研究中,采用壳聚糖低分子量(LCH)和壳聚糖中分子量(MCH)封装富含绿原酸和二咖啡酰奎尼酸(DCQAs)的yarrow提取物,该提取物对结肠腺癌细胞具有抗增殖活性。通过使用两种不同的技术:离子凝胶化和喷雾干燥进行CH微米/纳米颗粒的设计以增加提取物结肠递送。获得的离子凝胶化纳米粒子比喷雾干燥微粒更小,并且具有更高的产率值,但喷雾干燥微粒在包封效率(EE)方面表现最佳(>94%),还允许包含更大量的提取物。使用LCH设计的喷雾干燥微粒的LCH:提取物比例为6:1(1.25mg/mL),平均直径为1.31±0.21µm,EE值>93%,对于所有研究的酚类化合物。该制剂中包含的酚类化合物的释放曲线,在胃肠道pH值(2和7.4),向他们中的大多数展示了一个小的初始版本,随后在1小时增加,绿原酸在3小时时呈现较高的释放值(在pH2时为56.91%;在pH7.4时为44.45%)。3小时的DCQAs释放范围在9.01-40.73%之间,1,5-和3,4-DCQA更高。胃肠消化后,67.65%的绿原和大多数DCQAs保持包封。因此,可以提出喷雾干燥微粒作为一种有前途的载体,以增加先前描述为针对结肠直肠癌的潜在药剂的yarrow酚类化合物(主要是绿原酸和DCQAs)的结肠递送。
    In this study, chitosan low molecular weight (LCH) and chitosan medium molecular weight (MCH) were employed to encapsulate a yarrow extract rich in chlorogenic acid and dicaffeoylquinic acids (DCQAs) that showed antiproliferative activity against colon adenocarcinoma cells. The design of CH micro/nanoparticles to increase the extract colon delivery was carried out by using two different techniques: ionic gelation and spray drying. Ionic gelation nanoparticles obtained were smaller and presented higher yields values than spray-drying microparticles, but spray-drying microparticles showed the best performance in terms of encapsulation efficiency (EE) (> 94%), also allowing the inclusion of a higher quantity of extract. Spray-drying microparticles designed using LCH with an LCH:extract ratio of 6:1 (1.25 mg/mL) showed a mean diameter of 1.31 ± 0.21 µm and EE values > 93%, for all phenolic compounds studied. The release profile of phenolic compounds included in this formulation, at gastrointestinal pHs (2 and 7.4), showed for most of them a small initial release, followed by an increase at 1 h, with a constant release up to 3 h. Chlorogenic acid presented the higher release values at 3 h (56.91% at pH 2; 44.45% at pH 7.4). DCQAs release at 3 h ranged between 9.01- 40.73%, being higher for 1,5- and 3,4-DCQAs. After gastrointestinal digestion, 67.65% of chlorogenic and most DCQAs remained encapsulated. Therefore, spray-drying microparticles can be proposed as a promising vehicle to increase the colon delivery of yarrow phenolics compounds (mainly chlorogenic acid and DCQAs) previously described as potential agents against colorectal cancer.
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  • 文章类型: Journal Article
    在中医中,枸杞具有丰富的药用价值,和它的多糖是特别有趣的,由于其显著的药理作用和潜在的健康益处。本研究通过研究枸杞多糖(LBPs)与TLR4/MD-2复合物的相互作用以及胃肠道消化对这些相互作用的影响,研究了它们的免疫调节作用。我们发现LBP对TLR4/MD-2的亲和力结合及其细胞因子诱导能力受分子量的影响,具有中等大小的LBP(100-300kDa),具有更强的结合亲和力和诱导能力。相反,小于10kDa的LBP显示出降低的活性。此外,发现LBP级分中阿拉伯糖和半乳糖的含量与受体亲和力和细胞因子分泌均呈正相关。模拟的胃肠消化导致LBP降解为富含葡萄糖的较小片段。尽管这些片段对TLR4/MD-2复合物的结合亲和力降低,它们保持其促进细胞因子产生的活性。我们的发现强调了分子量和特定单糖组成在LBP免疫调节功能中的重要性,并强调了胃肠道消化对LBP作用的影响。这项研究有助于更好地了解中药多糖免疫调节作用的潜在机制及其实际应用。
    In traditional Chinese medicine, Lycium barbarum is of rich medicinal value, and its polysaccharides are particularly interesting due to their significant pharmacological effects and potential health benefits. This study investigated the immunomodulatory effects of Lycium barbarum polysaccharides (LBPs) by examining their interaction with the TLR4/MD-2 complex and the impacts of gastrointestinal digestion on these interactions. We discovered that the affinity binding of LBPs for TLR4/MD-2 and their cytokine induction capability are influenced by molecular weight, with medium-sized LBPs (100-300 kDa) exhibiting stronger binding affinity and induction capability. Conversely, LBPs smaller than 10 kDa showed reduced activity. Additionally, the content of arabinose and galactose within the LBPs fractions was found to correlate positively with both receptor affinity and cytokine secretion. Simulated gastrointestinal digestion resulted in the degradation of LBPs into smaller fragments that are rich in glucose. Although these fragments exhibited decreased binding affinity to the TLR4/MD-2 complex, they maintained their activity to promote cytokine production. Our findings highlight the significance of molecular weight and specific monosaccharide composition in the immunomodulatory function of LBPs and emphasize the influence of gastrointestinal digestion on the effects of LBPs. This research contributes to a better understanding of the mechanisms underlying the immunomodulatory effects of traditional Chinese medicine polysaccharides and their practical application.
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  • 文章类型: Journal Article
    葡聚糖是在由从特定物种和菌株的微生物剂获得的酶催化的反应中合成的胞外多糖。具有不同分子量的葡聚糖多糖的产物适用于多种药物和临床用途。葡聚糖溶液具有多种特性,包括粘度,溶解度,流变学,和热性能;因此,葡聚糖已经在几个领域进行了商业应用研究。某些细菌可产生不同分子量和构型的胞外多糖葡聚糖。不同分子量的葡聚糖产品已用于多个行业,包括医学,化妆品,和食物。本文旨在概述葡聚糖在输血和临床研究中的应用及其生物合成的报道。已经总结了有关从蔗糖生物合成葡聚糖的酶催化反应以及高分子量葡聚糖分子的生物转化过程以获得不同分子量和构型的制剂的信息。
    Dextran is an exopolysaccharide synthesized in reactions catalyzed by enzymes obtained from microbial agents of specific species and strains. Products of dextran polysaccharides with different molecular weights are suitable for diverse pharmaceutical and clinical uses. Dextran solutions have multiple characteristics, including viscosity, solubility, rheological, and thermal properties; hence, dextran has been studied for its commercial applications in several sectors. Certain bacteria can produce extracellular polysaccharide dextran of different molecular weights and configurations. Dextran products of diverse molecular weights have been used in several industries, including medicine, cosmetics, and food. This article aims to provide an overview of the reports on dextran applications in blood transfusion and clinical studies and its biosynthesis. Information has been summarized on enzyme-catalyzed reactions for dextran biosynthesis from sucrose and on the bio-transformation process of high molecular weight dextran molecules to obtain preparations of diverse molecular weights and configurations.
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  • 文章类型: Journal Article
    在这项研究中,纤溶蛋白酶是从围牛皮中分离纯化的,并对提取工艺进行了优化。酶的特性,如氨基酸组成,热稳定性,最佳温度,pH值,被调查了。戒毒后,通过硫酸铵沉淀浓缩从新鲜的Clamworm(PerinereisaibuhitensisGrub)收集的蛋白质。使用凝胶过滤树脂(SephadexG-100)纯化粗蛋白酶,阴离子交换树脂(DEAE-SepharoseFF),和疏水树脂(苯基琼脂糖6FF)。通过聚丙烯酰胺凝胶电泳(SDS-PAGE)测定蛋白酶的分子量。确定了蛋白酶的最适温度和最适pH。40-60%盐析段的粗蛋白酶活性最高,达到467.53U/mg。纯化粗蛋白的最佳工艺涉及DEAE-SepharoseFF和苯基Sepharose6FF的应用,这导致分离出一种名为Asp60-D1-P1的具有最高纤溶活性的蛋白酶;此外,酶活性为3367.76U/mg。通过Native-PAGE和SDS-PAGE分析表明,Asp60-D1-P1的分子量为44.5kDa,由分子量分别为6.5和37.8kDa的两个亚基组成,分别。Asp60-D1-P1的最佳温度为40°C,最佳pH值为8.0。
    In this study, fibrinolytic protease was isolated and purified from Perinereis aibuhitensis Grub, and the extraction process was optimized. The properties of the enzyme, such as the amino acid composition, thermal stability, optimal temperature, and pH, were investigated. After detoxification, proteins collected from fresh Clamworm (Perinereis aibuhitensis Grub) were concentrated via ammonium sulfate precipitation. The crude protease was purified using gel filtration resin (Sephadex G-100), anion exchange resin (DEAE-Sepharose FF), and hydrophobic resin (Phenyl Sepharose 6FF). The molecular weight of the protease was determined by polyacrylamide gel electrophoresis (SDS-PAGE). The optimum temperature and optimum pH of the protease were determined. The activity of crude protease in the 40-60% salt-out section was the highest, reaching 467.53 U/mg. The optimal process for purifying crude protein involved the application of DEAE-Sepharose FF and Phenyl Sepharose 6FF, which resulted in the isolation of a single protease known as Asp60-D1-P1 with the highest fibrinolytic activity; additionally, the enzyme activity was measured at 3367.76 U/mg. Analysis by Native-PAGE and SDS-PAGE revealed that the molecular weight of Asp60-D1-P1 was 44.5 kDa, which consisted of two subunits with molecular weights of 6.5 and 37.8 kDa, respectively. The optimum temperature for Asp60-D1-P1 was 40°C, and the optimal pH was 8.0.
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