Purification

净化
  • 文章类型: Journal Article
    海洋生物代表了有前途的生物活性肽资源,具有多种生物活性,如抗氧化剂,抗菌,抗高血压药,抗疲劳,和免疫调节活动。尽管有许多关于海洋生物活性肽的研究,关于海洋生物活性肽的生产技术和生物学应用的新兴趋势,缺乏全面的评论文章。在这次审查中,我们总结了与海洋生物活性肽有关的主要研究和发现,涵盖了他们生产的各个方面,净化,生物活动,基于纳米技术的战略,以及它们的潜在应用。目前,酶水解是生产海洋生物活性肽的最常用方法;下游纯化过程通常包括多种纯化技术的组合。由于它们不同的生物学特性,海洋肽作为食品中的活性成分在工业应用中引起了相当大的兴趣,Pharmaceutical,和化妆品行业。此外,纳米乳液等封装策略的掺入,纳米脂质体,和微乳液有望显着提高海洋肽的生物利用度和生物活性。未来的研究还应优先考虑通过体外和体内动物模型对海洋肽的潜在健康益处进行系统鉴定和验证。随着人体临床试验的进行。
    Marine organisms represent promising bioactive peptide resources with diverse biological activities such as antioxidant, antimicrobial, antihypertensive, anti-fatigue, and immunoregulatory activities. Despite many studies on marine bioactive peptides, there is a dearth of comprehensive review articles on the emerging trends that encompass the production techniques and the biological applications of marine bioactive peptides. In this review, we summarize the major research and findings related to marine bioactive peptides, encompassing aspects of their production, purification, biological activities, nanotechnology-based strategies, and their potential applications. Enzymatic hydrolysis currently stands out as the most commonly used method for producing marine bioactive peptides; the downstream purification process often includes a combination of multiple purification techniques. Due to their diverse biological properties, marine peptides have garnered considerable interest for industrial applications as active ingredients in the food, pharmaceutical, and cosmetics industries. Additionally, the incorporation of encapsulation strategies such as nano emulsion, nanoliposome, and microemulsions holds promise for significantly enhancing the bioavailability and bioactivity of marine peptides. Future research should also prioritize the systematic identification and validation of the potential health benefits of marine peptides by both in vitro and in vivo animal models, along with the conduct of human clinical trials.
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  • 文章类型: Journal Article
    C反应蛋白(CRP)在人类非特异性急性期反应的诊断和监测中起着至关重要的作用。相比之下,大鼠CRP(rCRP)是一种非典型的急性期蛋白,具有独特的功能,例如可能无法触发补体系统和显著升高的基线血浆浓度。为了促进对这些独特特征的体外研究,获得高质量的纯rCRP是必不可少的。在这里,我们探索了rCRP纯化的各种策略,包括从大鼠血浆中直接分离和在原核和真核系统中重组表达。我们的研究优化了重组表达系统,以提高rCRP的分泌和纯化效率。与传统的净化方法相比,我们提出了一种在毕赤酵母系统中表达和纯化rCRP的简化和有效方法。这种改进的方法在rCRP纯化的效率和有效性方面提供了显着改进,从而促进rCRP的进一步结构和功能研究。
    C-reactive protein (CRP) plays a crucial role in the diagnosis and monitoring of the non-specific acute phase response in humans. In contrast, rat CRP (rCRP) is an atypical acute-phase protein that possesses unique features, such as a possible incapacity to trigger the complement system and markedly elevated baseline plasma concentrations. To facilitate in vitro studies on these unique characteristics, obtaining high-quality pure rCRP is essential. Here we explored various strategies for rCRP purification, including direct isolation from rat plasma and recombinant expression in both prokaryotic and eukaryotic systems. Our study optimized the recombinant expression system to enhance the secretion and purification efficiency of rCRP. Compared to traditional purification methods, we present a streamlined and effective approach for the expression and purification of rCRP in the Pichia pastoris system. This refined methodology offers significant improvements in the efficiency and effectiveness of rCRP purification, thereby facilitating further structural and functional studies on rCRP.
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  • 文章类型: Journal Article
    镧系元素收缩涉及f-block元素中原子半径的减小低于预期水平。在第16组元素中观察到类似的收缩。由于d电子的存在,Se的原子半径(117pm)略大于S的原子半径(104pm),与原子半径从O(73pm)到S的显着增加相比。这种镧系元素样的收缩有助于Se的强大抗氧化能力。在这里,我们报告了一种选择性氧化策略,利用Se的强抗氧化性能从Se原料中去除共存的窄带隙Te杂质。该策略选择性地将挥发性Te杂质氧化为残留在蒸发源中的非挥发性TeO2。而在热蒸发沉积过程中只有挥发性Se沉积到衬底上。这使得能够制造具有1.88eV的宽带隙的高纯度Se薄膜,非常适合室内光伏(IPV)的最佳带隙。所得的Se光伏在1000勒克斯室内照明下表现出20.1%的效率,表现优于市场主导的非晶硅和所有类型的无铅钙钛矿IPV。未封装的Se器件在环境大气中储存20,000小时后没有显示效率降解。
    The lanthanide contraction involves a reduction in atomic radius among f-block elements below the expected level. A similar contraction is observed in group-16 elements. The atomic radius of Se (117 pm) is slightly larger than that of S (104 pm) arising from the presence of d electrons, compared to the significant increase in atomic radius from O (73 pm) to S. This lanthanide-like contraction contributes to Se\'s robust oxidative resistance. Here we report a selective oxidation strategy utilizing Se\'s strong antioxidative property to remove coexisting narrow-bandgap Te impurities from Se feedstocks. This strategy selectively oxidizes volatile Te impurities into involatile TeO2 that remains in the evaporation source, while only volatile Se deposits onto the substrate during the thermal-evaporation deposition process. This enables the fabrication of high-purity Se films possessing a wide bandgap of 1.88 eV, ideally suited to the optimal bandgap for indoor photovoltaics (IPVs). The resulting Se photovoltaics exhibit an efficiency of 20.1% under 1000-lux indoor illumination, outperforming market-dominant amorphous silicon and all types of lead-free perovskite IPVs. Unencapsulated Se devices show no efficiency degradation after 20,000 hours of storage in ambient atmosphere.
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  • 文章类型: Journal Article
    近年来,能够产生胞外多糖(EPS)的嗜热细菌变得有吸引力。嗜热菌合成的EPS由于其尚未开发的结构和生物学特性而值得研究。在这项研究中,来自嗜热的EPS,纯化革兰氏阳性细菌分离株并测试其生物活性。总共筛选了一百七个嗜热细菌产生胞外多糖的能力。属于地芽孢杆菌的9个分离株,副芽孢杆菌,Aeribacillus,选择EPS产量最高的厌氧杆菌属,和纯化的EPS(20、61、74、76、78、89、106、134和261)用于生物活性研究。选定的嗜热细菌的EPS产量介于117和419mg/L之间。在测试的EPS中,图61、106和261在最终浓度为1.5mg/mL时显示了对粪肠球菌JH2-2的抗菌作用。EPS样品具有显著的抗氧化能力,尤其是EPS134,在5mg/mL浓度下DPPH自由基清除活性最高,为100%,还原力最强。EPS20以31%的比率表现出最高的脂质过氧化抑制作用。当与标准阿卡波糖相比时,EPS显示弱的α-淀粉酶抑制活性。发现EPSs20、61、76、89、134和261的益生元指数高于菊粉,研究中所有测试乳酸菌的代表性益生元碳水化合物。所有检查的EPS均抑制由各种细菌形成的生物膜,具体取决于测试菌株。结果表明,嗜热EPS具有显著的抗氧化作用,益生元,和抗生物膜活性。因此,本研究中表征的EPS可能在健康和食品领域具有技术应用。
    Thermophilic bacteria able to produce exopolysaccharides (EPSs) have become attractive in recent years. EPSs synthesized by thermophiles are worth investigating due to their unexplored structural and biological properties. In this study, EPSs from thermophilic, Gram-positive bacterial isolates were purified and tested for their biological activities. A total of one hundred seven thermophilic bacteria were screened for their ability to produce exopolysaccharides. Nine isolates belonging to Geobacillus, Parageobacillus, Aeribacillus, and Anoxybacillus genera with highest EPS production were chosen, and purified EPSs (20, 61, 74, 76, 78, 89, 106, 134, and 261) were used for biological activity studies. EPS yields of selected thermophilic bacteria ranged between 117 and 419 mg/L. Among the tested EPSs, 61, 106, and 261 showed antibacterial effect against E. faecalis JH2-2 at a final concentration of 1.5 mg/mL. EPS samples had significant antioxidant capacity, especially EPS 134, with the highest DPPH radical scavenging activity of 100% at a concentration of 5 mg/mL and the strongest reducing power. EPS 20 showed the highest lipid peroxidation inhibition effect at a rate of 31%. EPSs displayed weak alpha-amylase inhibition activity when compared with standard acarbose. The prebiotic indices of EPSs 20, 61, 76, 89, 134, and 261 were found to be higher than that of inulin, a representative prebiotic carbohydrate for all tested lactic acid bacteria in the study. All examined EPSs inhibited the biofilms formed by various bacteria depending on the test strain. Results indicated that thermophilic EPSs had remarkable antioxidant, prebiotic, and antibiofilm activities. Therefore, EPSs characterized in this study may have technological applications in health and food fields.
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  • 文章类型: Journal Article
    植物单宁是环境友好的鞣剂。然而,它们通常赋予鞣制的皮革深色,并高度增加废水中的有机负荷。在这项研究中,我们分别对栗子单宁(CT)和硫酸化砂单宁(QT)采用纯化方案,以获得纯化的馏分(PCT和PQT)。这些样品通过GPC表征,1HNMR,13CNMR,FT-IR,和HPLC-DAD技术,并应用于鞣制试验。经由过程净化,非单宁成分和较小的分子如没食子酸,吡喃葡萄糖,从CT和QT中有效去除儿茶素,从而导致水分含量降低,pH值,和较浅的颜色纯化的馏分。用PCT和PQT处理的外壳皮革显示出理想的浅色阴影。此外,PCT和PQT制革废水中的有机负荷分别降低了13.5%和19.1%,分别,与传统CT和QT制革废水相比。此外,PCT和PQT处理的外壳皮革的物理和机械特性与CT和QT处理的外壳皮革相当。因此,植物单宁的纯化可以作为生产浅色植物鞣制皮革的可行策略,同时在植物鞣制过程中最大程度地减少有机污染物的排放。

    在线版本包含补充材料,可在10.1186/s42825-024-00171-9获得。
    Vegetable tannins are environmentally friendly tanning agents. However, they generally impart a dark colour to the tanned leather and highly contribute to the organic load in wastewaters. In this study, we employed a purification protocol separately on chestnut tannin (CT) and sulfited quebracho tannin (QT) to obtain the purified fractions (PCT and PQT). These samples were characterised by GPC, 1H NMR, 13C NMR, FT-IR, and HPLC-DAD techniques and applied for tanning tests. Through the purification process, non-tannin components and smaller molecules such as gallic acid, glucopyranose, and catechin were effectively removed from CT and QT, which consequently led to the reduced moisture content, pH value, and lighter colour of purified fractions. The crust leathers processed with PCT and PQT showed desirable light shades. Moreover, the organic loads in PCT and PQT tanning wastewater were reduced by 13.5% and 19.1%, respectively, when compared to those in traditional CT and QT tanning wastewater. Additionally, the physical and mechanical characteristics of crust leathers processed with PCT and PQT were comparable to those processed with CT and QT. Thus, purification of vegetable tannins may serve as a feasible strategy for producing light-colored vegetable-tanned leather while minimizing organic pollutant discharge during the vegetable tanning process.
    UNASSIGNED:
    UNASSIGNED: The online version contains supplementary material available at 10.1186/s42825-024-00171-9.
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  • 文章类型: Journal Article
    短链有机酸(SCOAs)是厌氧发酵过程中的中间体,可用于食品中,纺织品,和制药行业生产不同的最终用途产品。SCOA可以分开,纯化,集中在不同的过程中,比如蒸馏,提取或膜为基础的系统。SCOAs生产通过将这些可销售的酸作为高度浓缩的混合物与其他炼油厂工艺整合为酸性发酵工艺增加了更有利可图的可能性。本研究研究了两种回收SCOA的方法:i)通过微滤(MF)生产澄清的SCOA液体,然后通过反渗透(RO)进行浓缩,以及ii)通过所谓的集成中和和浓缩进行回收和浓缩。酸化反应方法。MF的结果表明,一些SCOA与固体一起保留在渗余物中。然而,在接下来的RO处理中,SCOA可以成功地浓缩,从SCOA液体中的产率保留超过90%。在后一种方法中,随着SCOAs总浓度从23g/L增加到146g/L,获得了无颜色的SCOAs液体。
    Short-chain organic acids (SCOAs) are the intermediates in the anaerobic fermentation process, and can be used in food, textile, and pharmaceutical industries to produce different end use products. SCOAs can be separated, purified, and concentrated by different processes, such as distillation, extraction or membrane-based systems. SCOAs production adds more profitable possibilities to an acidic fermentation process by integration these marketable acids as highly concentrated mixtures with other refinery processes. The present study investigated two approaches for recovering of SCOAs: i) the production of clarified SCOAs liquid by microfiltration (MF) and then performing their concentration by reverse osmosis (RO) and ii) the recovery and concentration by the so-called integrated neutralization and acidified reaction method. The results of MF showed that some SCOAs were retained in the retentate together with the solids. However, in the following RO treatment, SCOAs could be successfully concentrated with a yield retention of over 90 % from the SCOAs liquid. In the latter method, a color-free SCOAs liquid was obtained with an increase in the total SCOAs concentration from 23 g/L to 146 g/L.
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  • 文章类型: Journal Article
    作为一种自然广泛存在的饮食,化妆品,和治疗类黄酮,近年来,山奈酚的营养和药物特性得到了广泛的考虑。尽管已经进行了大量与山奈酚分析研究的不同方面相关的研究,人们感到缺乏对该化合物报道的各种方法和其他植物来源的全面总结,尤其是许多生物应用。这项研究,旨在提供由来源(植物物种)和与天然类黄酮(山奈酚)精确相关的分析信息组成的详细汇编。分析研究有一种趋势,支持现代生态友好型仪器和方法的应用。总之,超声辅助提取(UAE)是目前广泛用于提取山奈酚的最通用的先进方法。近年来,通过不同方法鉴定山奈酚的趋势越来越大。
    As a naturally widely-occurring dietary, cosmetic, and therapeutic flavonoid, kaempferol has gained much consideration for its nutritional and pharmaceutical properties in recent years. Although there have been performed a high number of studies associated with different aspects of kaempferol\'s analytical investigations, the lack of a comprehensive summary of the various methods and other plant sources that have been reported for this compound is being felt, especially for many biological applications. This study, aimed to provide a detailed compilation consisting of sources (plant species) and analytical information that was precisely related to the natural flavonoid (kaempferol). There is a trend in analytical research that supports the application of modern eco-friendly instruments and methods. In conclusion, ultrasound-assisted extraction (UAE) is the most general advanced method used widely today for the extraction of kaempferol. During recent years, there is an increasing tendency towards the identification of kaempferol by different methods.
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  • 文章类型: Journal Article
    海带是一种养殖广泛的优质海藻资源,具有良好的应用前景。在这项研究中,洋带岩藻聚糖(LF)是从洋带中提取的,并使用DEAE-SepharoseFastFlow凝胶柱纯化以获得四个不同等级。其中,LF4(Mw:165kDa),主要由岩藻糖(56.80%)组成,总糖含量最高(66.91%),硫酸盐含量最高(17.07%)。FT-RT和NMR结果表明,LF4主要由半乳糖化半乳糖岩藻糖组成,并且具有连接到岩藻糖C4的硫酸基。通过动物实验,结果表明,高脂血症小鼠的TC水平明显较高(5.52mmol/L),TG(2.28mmol/L)和LDL-C(5.12mmol/L)水平显著降低HDL-C(2mmol/L)。然而,LF具有调节高脂血症引起的脂质代谢紊乱的功效。以及调节血清中胆固醇转运的能力。此外,它调节了AMPK/ACC,PPAR-α/LAXRa,Nrf2/Nqo1、TLR4/NF-κB信号通路基因及蛋白在肝脏中的表达.此外,它促进了有益的短链脂肪酸(SCFA)的产生,同时改善了肠道菌群的组成和结构,包括平衡拟杆菌的丰度,Firmicutes,Muribaculaceae,Alloprevotella,大肠杆菌志贺氏菌,Prevotella和NK4A136。结果清楚地表明,LF4可以显着改善高脂血症,表明其作为功能性食品的应用前景。
    Laminaria digitata is a high-quality seaweed resource that is widely cultured and has good application prospects. In this study, Laminaria digitata fucoidan (LF) was extracted from Laminaria digitata, and purified using DEAE-Sepharose Fast Flow gel column to obtain four different grades. Among those, LF4 (Mw:165 kDa), mainly composed of fucose(56.80 %), had the highest total sugar (66.91 %) and sulfate (17.07 %) content. FT-RT and NMR results showed that LF4 was mainly composed of galactosylated galactofucose, and has a sulfate group attached to fucose C4. With the animal experimentation, it was revealed that hyperlipidaemic mice had significantly higher levels of TC (5.52 mmol/L), TG (2.28 mmol/L) and LDL-C (5.12 mmol/L) and significantly lower levels of HDL-C (2 mmol/L). However, LF had the efficacy in modulating the lipid metabolism disturbances induced by hyperlipidemia, as well as the ability to regulate cholesterol transport in serum. Moreover, it regulated AMPK/ACC, PPAR-α/LAXRa, Nrf2/Nqo1, TLR4/NF-κB signaling pathway genes and proteins expression in the liver. In addition, it promoted the production of beneficial short-chain fatty acids (SCFAs) while improving the composition and structure of gut microbiota, including balancing the abundance of Bacteroidota, Firmicutes, Muribaculaceae, Alloprevotella, Escherichia-Shigella, Prevotella and NK4A136. The results clearly indicated that LF4 could significantly ameliorate hyperlipidemia, suggesting its prospective application as a functional food.
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  • 文章类型: Journal Article
    通过将荞麦胰蛋白酶抑制剂(BTI)共价固定在表氯醇激活的交联琼脂糖凝胶(SelfinoseCL6B)上来制备胰蛋白酶亲和材料。激活自费糖CL6B的最佳条件是15%表氯醇和0.8MNaOH在40°C下2h。固定BTI的最佳pH为9.5。BTI-硒糖CL6B显示出2.25mg胰蛋白酶/(g载体)的最大吸附能力。该材料还显示出良好的可重用性,在30个循环后保留其初始吸附容量的90%以上。使用BTI-SelfinoseCL6B通过一步亲和层析从蝗虫匀浆中获得高纯度胰蛋白酶。以N-苯甲酰基-DL-精氨酸-硝基苯胺为底物,测定蝗虫胰蛋白酶的分子量和Km值为27kDa和0.241mM。胰蛋白酶活性的最佳温度和pH分别为55°C和9.0。该酶在30-50°C的温度范围和4.0-10.0的pH范围内表现出良好的稳定性。BTI-SelfinoseCL6B显示了在制备高纯度胰蛋白酶中的潜在应用,并发现了来自各种物种的更多新型胰蛋白酶。
    A trypsin affinity material was prepared by covalently immobilizing buckwheat trypsin inhibitor (BTI) on epichlorohydrin-activated cross-linked agarose gel (Selfinose CL 6 B). The optimal conditions for activating Selfinose CL 6 B were 15 % epichlorohydrin and 0.8 M NaOH at 40 °C for 2 h. The optimal pH for immobilizing BTI was 9.5. BTI-Sefinose CL 6 B showed a maximum adsorption capacity of 2.25 mg trypsin/(g support). The material also displayed good reusability, retaining over 90 % of its initial adsorption capacity after 30 cycles. High-purity trypsin was obtained from locust homogenate using BTI-Selfinose CL 6 B through one-step affinity chromatography. The molecular mass and Km value of locust trypsin were determined as 27 kDa and 0.241 mM using N-benzoyl-DL-arginine-nitroanilide as substrate. The optimal temperature and pH of trypsin activity were 55 °C and 9.0, respectively. The enzyme exhibited good stability in the temperature range of 30-50 °C and pH range of 4.0-10.0. BTI-Selfinose CL 6 B demonstrates potential application in the preparation of high-purity trypsin and the discovery of more novel trypsin from various species.
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  • 文章类型: Journal Article
    在这项研究中,茶多酚氧化酶(PPO)通过使用深共熔溶剂(DES)代替叔丁醇的三相分配(TPP)纯化。首先,对13种合成DES的性质进行了表征,选择DES-7(百里酚/十二烷酸)作为最佳替代溶剂。采用响应面法对工艺参数进行了优化。实验结果表明,当(NH4)2SO4浓度,DES与粗提物的比例,提取时间,pH为41%,0.5:1,75分钟,和5.6,分别茶多酚的回收率和纯化倍数分别为78.44%和8.26。采用SDS-PAGE和天然-PAGE对TTP系统纯化前后的PPO进行分析,并检测了PPO的分子量和纯化效果。此外,DES可以回收利用。结果表明,一种环境友好且稳定的DES,为大规模应用TPP提取PPO提供参考。
    In this study, tea polyphenol oxidase (PPO) was purified via three-phase partitioning (TPP) using a deep eutectic solvent (DES) instead of t-butanol. First, the properties of 13 types of synthesized DESs were characterized, and DES-7 (thymol/dodecanoic acid) was selected as the best alternative solvent. The process parameters were optimized using response surface methodology. The experimental results revealed that when the (NH4)2SO4 concentration, DES to crude extract ratio, extraction time, and pH were 41%, 0.5:1, 75 min, and 5.6, respectively, the recovery and purification fold of tea PPO were 78.44% and 8.26, respectively. SDS-PAGE and native-PAGE were used to analyze the PPO before and after purification of the TTP system, and the molecular weight and purification effect of PPO were detected. Moreover, the DES could be recovered and recycled. The results indicate an environmentally friendly and stable DES, and provide a reference for the large-scale application of TPP to extract PPO.
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