Escherichia coli (E. coli)

大肠杆菌 (E.大肠杆菌)
  • 文章类型: Journal Article
    用于从采矿废水中回收稀土元素(REE)的微生物表明有痕量的大肠杆菌(E.大肠杆菌2149.6μg/g),球形芽孢杆菌(1636.6μg/g),芽孢杆菌(1469.3μg/g),和蜡样芽孢杆菌(1083.9μg/g)。其中,大肠杆菌对REE比非REE(Mn和Zn)显示出亲和力。由于随着离子半径的减小,吸附过程增加,重稀土元素在大肠杆菌上的吸附量(1511.1μg/g)高于轻稀土元素(638.0μg/g)。此外,大肠杆菌在从采矿废水中回收稀土元素方面表现出稳定性,如4个周期所示。SEM-EDS,XPS和FTIR显示REE对细胞有破坏作用,在细胞表面吸收和解吸的稀土元素,包括与离子如Na+的离子交换,配体与官能团如-NH2结合。最后,成本评估证实了大肠杆菌从采矿废水中回收稀土元素的经济可行性。
    Microbes used for the recovery of rare earth elements (REEs) from mining wastewater indicated traces of Escherichia coli (E. coli, 2149.6 μg/g), Bacillus sphaericus (1636.6 μg/g), Bacillus mycoides (1469.3 μg/g), and Bacillus cereus (1083.9 μg/g). Of these, E. coli showed an affinity for REEs than non-REEs (Mn and Zn). The amount of heavy REEs adsorbed (1511.1 μg/g) on E. coli was higher than light REEs (638.0 μg/g) due to the process of increasing adsorption with decreasing ionic radius. Additionally, E. coli demonstrated stability in the recovery of REEs from mining wastewater, as evidenced by 4 cycles. SEM-EDS, XPS and FTIR showed that REEs had a disruptive effect on cells, REEs absorbed and desorbed on the cell surface including ion exchange with ions such as Na+, ligand binding with functional groups like -NH2. Finally, the cost assessment confirmed the economically feasible of E. coli in recovery of REEs from mining wastewater.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    目的:。尽管粘菌素作为最后的抗生素至关重要,有限的研究调查了柬埔寨人类感染中粘菌素的耐药性。本研究旨在调查2016年至2020年在柬埔寨分离的产超广谱β-内酰胺酶(ESBL)和产碳青霉烯酶(CP)肺炎克雷伯菌(KP)和大肠杆菌(EC)的粘菌素耐药性及其分子决定因素。
    方法:。通过肉汤微量稀释测试EC(n=223)和KP(n=39)的粘菌素最小抑制浓度(MIC)。对抗性分离株进行PCR,以检测两组分系统(TCS)中的移动粘菌素抗性基因(mcr)和染色体突变。
    结果:。18个分离株(10KP,8EC)显示,在ESBL分离株中,EC中的粘菌素抗性为5.9%,KP中的粘菌素抗性为34.8%,CP分离株中EC为1%,KP为12.5%。耐药性与mcr变异体相关(13/18分离株,mcr-1,mcr-3和mcr-8.2)和EC和KP内的TCS突变,随着在柬埔寨首次发现mcr-8.2,在EC的TCS中发现可能与粘菌素抗性相关的新突变(PhoPI47V,PhoQN352K,PmrBG19R,PmrDG85R)以及mcr基因和粘菌素抗性的共同出现,赋予了11/18分离株的TCS突变。
    结论:研究结果强调了柬埔寨人类感染涉及的ESBL-和CP-肠杆菌科中粘菌素耐药性的存在,以及TCS中的染色体突变以及EC和KP中mcr-8.2的出现。它强调了持续监测的必要性,抗菌药物管理,和控制措施,以减轻粘菌素耐药性的传播。
    OBJECTIVE: Despite the critical importance of colistin as a last-resort antibiotic, limited studies have investigated colistin resistance in human infections in Cambodia. This study aimed to investigate the colistin resistance and its molecular determinants among Extended-spectrum beta-lactamase (ESBL)- and carbapenemase-producing (CP) Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) isolated in Cambodia between 2016 and 2020.
    METHODS: E. coli (n = 223) and K. pneumoniae (n = 39) were tested for colistin minimum inhibitory concentration (MIC) by broth microdilution. Resistant isolates were subjected to polymerase chain reaction (PCR) for detection of mobile colistin resistance genes (mcr) and chromosomal mutations in the two-component system (TCS).
    RESULTS: Eighteen isolates (10 K. pneumoniae and 8 E. coli) revealed colistin resistance with a rate of 5.9% in E. coli and 34.8% in K. pneumoniae among ESBL isolates, and 1% in E. coli and 12.5% in K. pneumoniae among CP isolates. The resistance was associated with mcr variants (13/18 isolates, mcr-1, mcr-3, and mcr-8.2) and TCS mutations within E. coli and K. pneumoniae, with the first detection of mcr-8.2 in Cambodia, the discovery of new mutations potentially associated to colistin resistance in the TCS of E. coli (PhoP I47V, PhoQ N352K, PmrB G19R, and PmrD G85R) and the co-occurrence of mcr genes and colistin resistance conferring TCS mutations in 11 of 18 isolates.
    CONCLUSIONS: The findings highlight the presence of colistin resistance in ESBL- and CP- Enterobacteriaceae involved in human infections in Cambodia as well as chromosomal mutations in TCS and the emergence of mcr-8.2 in E. coli and K. pneumoniae. It underscores the need for continuous surveillance, antimicrobial stewardship, and control measures to mitigate the spread of colistin resistance.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    联合下水道溢流(CSO)将微生物污染物引入接收水体,从而对公众健康构成风险。本研究系统地研究了选择大肠杆菌(E。大肠杆菌)作为目标微生物污染物。与UV、PAA,和UV/H2O2工艺。增加PAA用量大大提高了消毒效率,而浊度和有机物阻碍了失活性能。单线态氧(1O2),羟基(·OH)和有机自由基(RO·)有助于大肠杆菌的失活,·OH和RO·起着突出的作用。细胞内活性氧的变化,丙二醛,酶活性,大肠杆菌细胞的DNA含量和生化成分表明,UV/PAA主要引起细胞内分子的氧化损伤,而不是细胞膜脂质的损伤。因此有效地限制了大肠杆菌的再生长。此外,UV/PAA工艺在消毒实际原始公民社会组织方面表现突出,在反应4分钟后实现总细菌的2.90-log失活。这些结果突出了UV/PAA工艺在CSO消毒中的实际适用性和有效性。
    Combined sewer overflows (CSOs) introduce microbial contaminants into the receiving water bodies, thereby posing risks to public health. This study systematically investigated the disinfection performance and mechanisms of the combined process of ultraviolet and peracetic acid (UV/PAA) in CSOs with selecting Escherichia coli (E. coli) as a target microbial contaminant. The UV/PAA process exhibited superior performance in inactivating E. coli in simulated CSOs compared with UV, PAA, and UV/H2O2 processes. Increasing the PAA dosage greatly enhanced the disinfection efficiency, while turbidity and organic matter hindered the inactivation performance. Singlet oxygen (1O2), hydroxyl (•OH) and organic radicals (RO•) contributed to the inactivation of E. coli, with •OH and RO• playing the prominent role. Variations of intracellular reactive oxygen species, malondialdehyde, enzymes activities, DNA contents and biochemical compositions of E. coli cells suggested that UV/PAA primarily caused oxidative damage to intracellular molecules rather than the damage to the lipids of the cell membrane, therefore effectively limited the regrowth of E. coli. Additionally, the UV/PAA process displayed an outstanding performance in disinfecting actual raw CSOs, achieving a 2.90-log inactivation of total bacteria after reaction for 4 min. These results highlighted the practical applicability and effectiveness of the UV/PAA process in the disinfection of CSOs.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    城市河流为水上娱乐提供了绝佳的机会。这项研究以概率方式评估了比滩风景区城市河流中与水上娱乐相关的健康风险,台湾,通过采用定量微生物风险评估和残疾调整寿命年(DALYs)。此外,城市化对水源性致病性大肠杆菌(E.大肠杆菌)进行了调查。首先,收集了比滩风景区和上游河段的河流大肠杆菌含量数据,模型参数是通过对河流休闲者进行问卷调查获得的。然后采用蒙特卡罗模拟来解决参数不确定性。最后,计算DALY以量化潜在寿命损失和残疾年限方面的累积影响。结果表明,皮划艇的疾病负担(DB)的90%置信区间为每人每年0.2-74.1×10-6,0.01-94.0×10-6和0.3-128.9×10-6DALY(pppy),游泳,钓鱼,分别,在比坦风景区。此外,比坦风景区附近的城市化将上游农村地区的河流娱乐者的DB风险增加了大约一倍。在第95百分位数,DB风险超过了世界卫生组织(1×10-6)或美国环境保护局(1×10-4)建议的容忍度。研究结果表明,同时实施污水下水道系统和最佳管理实践可以将河流休闲者的健康风险降低至少一半,将DALY水平降低到1×10-4以下,甚至1×10-5pppy。
    Urban rivers provide an excellent opportunity for water recreation. This study probabilistically assessed health risks associated with water recreation in urban rivers in the Bitan Scenic Area, Taiwan, by employing quantitative microbial risk assessment and disability-adjusted life years (DALYs). Moreover, the effects of urbanization on the health risks of river recreation induced by waterborne pathogenic Escherichia coli (E. coli) were investigated. First, data on river E. coli levels were collected in both the Bitan Scenic Area and the upstream river section, and model parameters were obtained through a questionnaire administered to river recreationists. Monte Carlo simulation was then employed to address parameter uncertainty. Finally, DALYs were calculated to quantify the cumulative effects in terms of potential life lost and years lived with disability. The results indicated that the 90 % confidence intervals for the disease burden (DB) were 0.2-74.1 × 10-6, 0.01-94.0 × 10-6, and 0.3-128.9 × 10-6 DALY per person per year (pppy) for canoeing, swimming, and fishing, respectively, in the Bitan Scenic Area. Furthermore, urbanization near the Bitan Scenic Area approximately doubled the DB risks to river recreationists in upstream rural areas. At the 95th percentile, the DB risks exceeded the tolerances recommended by the World Health Organization (1 × 10-6) or U.S. Environmental Protection Agency (1 × 10-4). The findings suggest that the simultaneous implementation of effluent sewer systems and best management practices can reduce health risks to river recreationists by at least half, reducing the DALY levels below 1 × 10-4 or even 1 × 10-5 pppy.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    细胞色素bo3喹啉氧化酶属于血红素-铜氧化还原酶(HCO)超家族,这是呼吸链的一部分,对细胞存活至关重要。虽然在过去的几十年中已经对cytbo3的反应机理进行了广泛的研究,由于缺乏结构信息,有关其底物结合和产品释放的具体细节仍未阐明。这里,我们报告了组装在肽盘中的大肠杆菌的cytbo3的2.8进行低温电子显微镜结构。我们的结构模型显示亚基I的氨基酸1-41的构象与所有先前公开的结构不同,而该酶的其余部分相似。我们的新构象显示出与跨膜螺旋相反的“U形”组装,名为\"TM0\",在其他报告的结构模型中。然而,TM0阻断泛醌-8(反应产物)的释放,表明其他cytbo3构象应该存在。我们的结构模型为促进底物/产品交换的“开放”构象提供了实验证据。这项工作有助于进一步了解这种氧化酶的反应周期,这可能有利于潜在的药物/抗生素的健康科学设计。
    Cytochrome bo3 quinol oxidase belongs to the heme‑copper-oxidoreductase (HCO) superfamily, which is part of the respiratory chain and essential for cell survival. While the reaction mechanism of cyt bo3 has been studied extensively over the last decades, specific details about its substrate binding and product release have remained unelucidated due to the lack of structural information. Here, we report a 2.8 Å cryo-electron microscopy structure of cyt bo3 from Escherichia coli assembled in peptidiscs. Our structural model shows a conformation for amino acids 1-41 of subunit I different from all previously published structures while the remaining parts of this enzyme are similar. Our new conformation shows a \"U-shape\" assembly in contrast to the transmembrane helix, named \"TM0\", in other reported structural models. However, TM0 blocks ubiquinone-8 (reaction product) release, suggesting that other cyt bo3 conformations should exist. Our structural model presents experimental evidence for an \"open\" conformation to facilitate substrate/product exchange. This work helps further understand the reaction cycle of this oxidase, which could be a benefit for potential drug/antibiotic design for health science.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    与分子印迹聚合物(MIP)集成的基于距离的纸张分析设备(dPAD)的开发,以监测大肠杆菌(E.大肠杆菌)在食物样本中的水平。使用设计的亲水桥阀控制装置上的流体工作流。多巴胺用作在dPAD上形成大肠杆菌选择性MIP层的单体。检测原理依赖于大肠杆菌对铜(II)(Cu2)触发的邻苯二胺(OPD)在纸基材上的氧化的抑制作用。定量检测只是通过目测检测区OPD残留的黄色来确定,与大肠杆菌浓度成正比。该传感表现出25.0至1200.0CFUmL-1(R2=0.9992)的线性范围和用于大肠杆菌检测的25.0CFUmL-1的检测限(LOD)。此外,该技术是高度选择性的,即使是与OPD反应形成氧化OPD的分子也没有干扰。所开发的设备证明了食品样品中大肠杆菌定量的准确性和精密度,回收率在98.3和104.7%之间,最高相对标准偏差(RSD)为4.55%。T检验验证显示在我们的方法和商业测定之间测量的大肠杆菌浓度没有显著差异。所提出的dPAD传感器具有在食品样品中选择性和负担得起的大肠杆菌测定的潜力,而不需要样品制备。此外,该策略可以扩展到监测可以开发MIP并将其集成到纸-微流体平台中的其他分子。
    The development of distance-based paper analytical devices (dPADs) integrated with molecularly imprinted polymers (MIPs) to monitor Escherichia coli (E. coli) levels in food samples is presented. The fluidic workflow on the device is controlled using a designed hydrophilic bridge valve. Dopamine serves as a monomer for the formation of the E. coli-selective MIP layer on the dPADs. The detection principle relies on the inhibition of the E. coli toward copper (II) (Cu2+)-triggered oxidation of o-phenylenediamine (OPD) on the paper substrate. Quantitative detection is simply determined through visual observation of the residual yellow color of the OPD in the detection zone, which is proportional to E. coli concentration. The sensing exhibits a linear range from 25.0 to 1200.0 CFU mL-1 (R2 = 0.9992) and a detection limit (LOD) of 25.0 CFU mL-1 for E. coli detection. Additionally, the technique is highly selective with no interference even from the molecules that have shown to react with OPD to form oxidized OPD. The developed device demonstrates accuracy and precision for E. coli quantification in food samples with recovery percentages between 98.3 and 104.7% and the highest relative standard deviation (RSD) of 4.55%. T-test validation shows no significant difference in E. coli concentration measured between our method and a commercial assay. The proposed dPAD sensor has the potential for selective and affordable E. coli determination  in food samples without requiring sample preparation. Furthermore, this strategy can be extended to monitor other molecules for which MIP can be developed and integrated into paper-microfluidic platform.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

    求助全文

  • 文章类型: Journal Article
    引言从人临床样本中分离的细菌中的抗微生物耐药性(AMR)的巨大增加导致治疗失败。通过下一代测序(NGS)或全基因组测序(WGS)增加监测可以促进耐药菌株的流行病学研究,抗性基因,以及它们可能携带的其他毒力决定因素。方法本研究包括30个大肠杆菌(E.大肠杆菌)从Prathima医学科学研究所的尿路感染(UTI)患者中获得的分离株,Karimnagar,印度。鉴定了所有细菌分离株,和抗菌药物敏感性模式通过常规微生物技术确定,并通过自动化系统确认。使用NGS对所有分离株进行了调查,以鉴定编码抗性的基因,如超广谱β-内酰胺酶(ESBLs),金属-β-内酰胺酶,和毒力基因。多位点序列分型(MLST)用于了解流行的菌株类型,并进行血清分型以评估分离物携带的O(细胞壁抗原)和H(鞭毛抗原)血清型。结果常规药敏试验发现15株(50%)对亚胺培南(IPM)耐药,10例(33.33%)对阿米卡星(AK)耐药,13例(43.33%)对哌拉西林他唑巴坦(PTZ)耐药,17例(56.66%)对头孢菌素耐药,14例(46.66%)对呋喃妥因(NIT)耐药。在分离物中,26(86.66%)揭示了多种抗生素抗性基因的存在,证据表明至少有一种基因编码β-内酰胺酶抗性。blaCTX-M(19/30,63.33%)基因患病率较高,其次是blaTEM和blaOXA-1。在三个分离物中发现了blaNDM-5基因(3/30,10%)。本研究中鉴定的毒力基因是iutA,sat,ISS,还有爸爸,在其他人中。大肠杆菌血清型主要属于O25:H4(5,16.66%),其次是O102:H6(4,13.33%)。在所检查的样品中鉴定出总共16个MLST变体。在识别的基于MLST的序列类型(ST)中,以ST-131(7,23.33%)为主,其次是ST-167(3,10%)和ST-12(3,10%)。结论本研究结果表明,从UTI患者中分离的大肠杆菌菌株具有潜在的耐药性和毒力基因,属于基于MLST的不同菌株类型。使用分子分析(如NGS)仔细评估细菌菌株可以促进对细菌抗生素抗性及其毒力潜力的更好理解。这可以使医生选择合适的抗菌药物,并有助于更好的患者管理,从而防止耐药细菌的出现和传播。
    Introduction An enormous increase in antimicrobial resistance (AMR) among bacteria isolated from human clinical specimens contributed to treatment failures. Increased surveillance through next-generation sequencing (NGS) or whole genome sequencing (WGS) could facilitate the study of the epidemiology of drug-resistant bacterial strains, resistance genes, and other virulence determinants they are potentially carrying. Methods This study included 30 Escherichia coli (E. coli) isolates obtained from patients suffering from urinary tract infections (UTIs) attending Prathima Institute of Medical Sciences, Karimnagar, India. All bacterial isolates were identified, and antimicrobial susceptibility patterns were determined through conventional microbiological techniques and confirmed by automated systems. All the isolates were investigated using NGS to identify genes coding for resistance, such as extended-spectrum beta-lactamases (ESBLs), metallo-beta-lactamases, and virulence genes. Multilocus sequence typing (MLST) was used to understand the prevalent strain types, and serotyping was carried out to evaluate the type of O (cell wall antigen) and H (flagellar antigen) serotypes carried by the isolates. Results The conventional antimicrobial susceptibility testing revealed that 15 (50%) isolates were resistant to imipenem (IPM), 10 (33.33%) were resistant to amikacin (AK), 13 (43.33%) were resistant to piperacillin-tazobactam (PTZ), 17 (56.66%) were resistant to cephalosporins, and 14 (46.66%) were resistant to nitrofurantoin (NIT). Among the isolates, 26 (86.66%) had revealed the presence of multiple antibiotic-resistant genes with evidence of at least one gene coding for beta-lactamase resistance. There was a high prevalence of blaCTX-M (19/30, 63.33%) genes, followed by blaTEM and blaOXA-1. The blaNDM-5 gene was found in three isolates (3/30, 10%). The virulence genes identified in the present study were iutA, sat, iss, and papC, among others. The E. coli serotype found predominantly belonged to O25:H4 (5, 16.66%), followed by O102:H6 (4, 13.33%). A total of 16 MLST variants were identified among the examined samples. Of the MLST-based sequence types (STs) identified, ST-131 (7, 23.33%) was the predominant one, followed by ST-167 (3, 10%) and ST-12 (3, 10%). Conclusions The study results demonstrated that the E. coli strains isolated from patients suffering from UTIs potentially carried antimicrobial resistance and virulence genes and belonged to different strain types based on MLST. Careful evaluation of bacterial strains using molecular analyses such as NGS could facilitate an improved understanding of bacterial antibiotic resistance and its virulence potential. This could enable physicians to choose appropriate antimicrobial agents and contribute to better patient management, thereby preventing the emergence and spread of drug-resistant bacteria.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Case Reports
    急性胆管炎是由胆道梗阻引起的胆道感染引起的潜在危及生命的疾病。该病例报告强调了一位老年患者的急性胆管炎的异常表现,其特征是存在产超广谱β-内酰胺酶的大肠杆菌。我们旨在强调识别老年人群各种临床表现的重要性,以加强及时诊断和适当管理。该病例强调了更好地了解患者潜在致病生物的危险因素及其对选择适当抗生素和改善临床结果的敏感性的重要性。
    Acute cholangitis is a potentially life-threatening condition caused by an infection of the biliary tract resulting from biliary obstruction. This case report highlights an unusual presentation of acute cholangitis in an elderly patient characterized by the presence of extended-spectrum beta-lactamase-producing Escherichia coli. We aim to emphasize the significance of recognizing diverse clinical manifestations in the elderly population to enhance timely diagnosis and appropriate management. The case highlights the importance of better understanding patient risk factors for potential causative organisms and their susceptibility to selecting proper antibiotics and improving clinical outcomes.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    了解与海洋水污染升高风险相关的历史环境决定因素可以加强对加拿大海洋海滩的监测,这也可以为预测的海洋水质模型和正在进行的气候变化准备工作提供信息。本研究旨在评估最能预测大肠杆菌(E.大肠杆菌)在大温哥华公共海滩的浓度,不列颠哥伦比亚省,通过结合该地区2013年至2021年的微生物水质数据和公开可用的环境数据。我们开发了贝叶斯对数正态混合效应回归模型,以评估大温哥华地区15个海滩的几何大肠杆菌浓度的预测因子。我们发现,较高的几何平均大肠杆菌水平是由较高的前样本日大肠杆菌浓度预测,前48小时降雨量较高,以及在24小时平均紫外线(UV)指数的中值或更高水平处更高的24小时平均空气温度。相比之下,预计较高的平均盐度水平会导致较低的大肠杆菌水平。最后,我们确定预测因子的平均影响因海滩而异。我们的发现可以为建立实时预测的海洋水质模型奠定基础,以实现更及时的海滩管理决策。
    Understanding historical environmental determinants associated with the risk of elevated marine water contamination could enhance monitoring marine beaches in a Canadian setting, which can also inform predictive marine water quality models and ongoing climate change preparedness efforts. This study aimed to assess the combination of environmental factors that best predicts Escherichia coli (E. coli) concentration at public beaches in Metro Vancouver, British Columbia, by combining the region\'s microbial water quality data and publicly available environmental data from 2013 to 2021. We developed a Bayesian log-normal mixed-effects regression model to evaluate predictors of geometric E. coli concentrations at 15 beaches in the Metro Vancouver Region. We identified that higher levels of geometric mean E. coli levels were predicted by higher previous sample day E. coli concentrations, higher rainfall in the preceding 48 h, and higher 24-h average air temperature at the median or higher levels of the 24-h mean ultraviolet (UV) index. In contrast, higher levels of mean salinity were predicted to result in lower levels of E. coli. Finally, we determined that the average effects of the predictors varied highly by beach. Our findings could form the basis for building real-time predictive marine water quality models to enable more timely beach management decision-making.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

  • 文章类型: Journal Article
    金黄色葡萄球菌(S。金黄色葡萄球菌),革兰氏阳性细菌,引起广泛的感染,早期诊断具有挑战性。靶向麦芽糖糊精转运蛋白已成为对细菌成像的有希望的策略,并且已经能够对包括金黄色葡萄球菌在内的多种细菌成像。然而,对金黄色葡萄球菌的麦芽糊精转运蛋白知之甚少,这阻止了麦芽糊精转运蛋白的新的金黄色葡萄球菌特异性配体的开发。在革兰氏阳性菌中,包括金黄色葡萄球菌,麦芽糊精转运的第一步是麦芽糊精结合蛋白malE与麦芽糊精的结合。因此,了解金黄色葡萄球菌malE的结合亲和力和特性对于开发基于麦芽糊精的高效成像探针很重要。我们评估了金黄色葡萄球菌的malE对各种长度的麦芽糊精的亲和力。金黄色葡萄球菌的MalE(SAmalE)在大肠杆菌BL21(DE3)中表达并通过Ni-NTA树脂纯化。用等温滴定量热法评价SAmalE对麦芽糊精的亲和力。SAmalE对麦芽糖的亲和力低,但与麦芽三糖和更长的麦芽糊精结合,直至麦芽七糖,亲和力高达Ka=9.02±0.49×105M-1。SAmalE与麦芽三糖-麦芽七糖的结合是放热的并且符合单结合位点模型。SAmalE与麦芽三糖的结合反应中的Van\'tHoff焓为9.9±1.3kcal/mol,用麦芽四糖观察到SAmalE的最高亲和力,Ka=9.02±0.49×105M-1。在ΔH-T*ΔS的曲线图中,在SAmalE与麦芽糊精的结合反应中观察到焓熵补偿效应。阿卡波糖和麦芽四醇与SAmalE结合,表明SAmalE耐受麦芽糖糊精的还原和非还原末端上的修饰。我们的结果表明,与ECmalE不同,与链球菌的麦芽糊精结合蛋白相似,SAmalE主要通过氢键与麦芽糊精结合。这与链球菌的麦芽糊精结合蛋白不同,以高亲和力与麦芽四醇结合的SAmalE。了解麦芽糊精结合蛋白的结合特性和对麦芽糊精修饰的耐受性将有望为开发基于细菌物种特异性麦芽糊精的成像探针提供基础。
    Staphylococcus aureus (S. aureus), a Gram-positive bacterium, causes a wide range of infections, and diagnosis at an early stage is challenging. Targeting the maltodextrin transporter has emerged as a promising strategy for imaging bacteria and has been able to image a wide range of bacteria including S. aureus. However, little is known about the maltodextrin transporter in S. aureus, and this prevents new S. aureus specific ligands for the maltodextrin transporter from being developed. In Gram-positive bacteria, including S. aureus, the first step of maltodextrin transport is the binding of the maltodextrin-binding protein malE to maltodextrins. Thus, understanding the binding affinity and characteristics of malE from S. aureus is important to developing efficient maltodextrin-based imaging probes. We evaluated the affinity of malE of S. aureus to maltodextrins of various lengths. MalE of S. aureus (SAmalE) was expressed in E. coli BL21(DE3) and purified by Ni-NTA resin. The affinities of SAmalE to maltodextrins were evaluated with isothermal titration calorimetry. SAmalE has low affinity to maltose but binds to maltotriose and longer maltodextrins up to maltoheptaose with affinities up to Ka = 9.02 ± 0.49 × 105 M-1. SAmalE binding to maltotriose-maltoheptaose was exothermic and fit a single-binding site model. The van\'t Hoff enthalpy in the binding reaction of SAmalE with maltotriose was 9.9 ± 1.3 kcal/mol, and the highest affinity of SAmalE was observed with maltotetraose with Ka = 9.02 ± 0.49 × 105 M-1. In the plot of ΔH-T*ΔS, the of Enthalpy-Entropy Compensation effect was observed in binding reaction of SAmalE to maltodextrins. Acarbose and maltotetraiol bind with SAmalE indicating that SAmalE is tolerant of modifications on both the reducing and non-reducing ends of maltodextrins. Our results show that unlike ECmalE and similar to the maltodextrin binding protein of Streptococci, SAmalE primarily binds to maltodextrins via hydrogen bonds. This is distinct from the maltodextrin binding protein of Streptococci, SAmalE that binds to maltotetraiol with high affinity. Understanding the binding characteristics and tolerance to maltodextrins modifications by maltodextrin binding proteins will hopefully provide the basis for developing bacterial species-specific maltodextrin-based imaging probes.
    导出

    更多引用

    收藏

    翻译标题摘要

    我要上传

       PDF(Pubmed)

公众号