Biochemical characteristics

  • 文章类型: Journal Article
    空气污染是城市地区紧迫的环境问题,尤其是像德里这样人口稠密的城市,印度。然而,植物物种可以有效地捕获空气中的悬浮污染物。鉴于此,本研究旨在调查污染缓解潜力的季节性变化(季风前和季风后),生化特性,以及德里某些植物物种的悬浮颗粒物(SPM)捕获能力。此外,使用生化参数,植物形态学,和社会经济因素,该研究计算了容忍指数,如空气污染容忍指数(APTI)和预期绩效指数(API)。FicusreligiosaL.表现出最高的APTI值,为11.94,而Polyalthialongifolia(Sonn。)Thwaites在季风前期间显示了最低的7.99APTI值。无花果在叶片上显示出最大的SPM粘附力,沉积量为1305.46µg/cm2,而F.religiosa的最低SPM沉积量为56.62µg/cm2。此外,统计分析表明,抗坏血酸和叶绿素含量(R2>0.6)与APTI呈正相关。此外,F.religiosa在季风前表现出叶绿素含量与SPM沉积之间的显着Pearson相关性(P<0.05)。该研究强调了基于植物的空气污染缓解的动态性质。它为绿色基础设施作为解决城市环境中空气质量问题的可持续解决方案的潜力提供了宝贵的见解。结果强调了在制定城市绿化策略以应对空气污染时选择适当的植物物种并考虑季节性变化的重要性。
    Air pollution is a pressing environmental concern in urban areas, especially in densely populated cities like Delhi, India. However, plant species can effectively capture airborne suspended pollutants. Given this, the present study aimed to investigate the seasonal variations (pre- and post-monsoon) in the pollution-mitigating potential, biochemical characteristics, and suspended particulate matter (SPM) capturing capacities of select plant species in Delhi. Also, using biochemical parameters, plant morphology, and socioeconomic factors, the study computed tolerance indices such as the Air Pollution Tolerance Index (APTI) and Anticipated Performance Index (API). Ficus religiosa L. exhibited the highest APTI value of 11.94, while Polyalthia longifolia (Sonn.) Thwaites displayed the lowest 7.99 APTI value during the pre-monsoon. Ficus benghalensis L. showed the maximum SPM adhesion on the leaves, with a deposition of 1305.46 µg/cm2, whereas F. religiosa exhibited the lowest SPM deposition of 56.62 µg/cm2. Moreover, the statistical analysis indicated a positive correlation between ascorbic acid and chlorophyll content (R2 > 0.6) with APTI. Also, F. religiosa demonstrated a significant Pearson\'s correlation (P < 0.05) between chlorophyll content and SPM deposition during the pre-monsoon. The study highlighted the dynamic nature of plant-based air pollution mitigation. It offered valuable insights into the potential of green infrastructure as a sustainable solution for addressing air quality concerns in urban environments. The results emphasized the significance of selecting adequate plant species and considering seasonal variations in developing urban greening strategies to combat air pollution.
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  • 文章类型: Journal Article
    Cipangopaludinachinensis,作为中国经济上重要的物种,代表自然界中的腹足动物,由于猎物选择有限,经常遭受饥饿压力。然而,对抗饥饿的潜在反应机制尚未得到深入研究。我们收集了多次饥饿胁迫(0、7、30和60天)下的C.chinensis的营养,生化特征和转录组分析。结果表明:长期饥饿胁迫(>30天)导致蜗牛营养成分出现明显波动,随着体重的急剧减少,存活率和消化酶活性(淀粉酶,蛋白酶,和脂肪酶),并显着增强了蜗牛的抗氧化酶活性。比较转录组分析显示3538个差异表达基因(DEGs),与特定的饥饿应激反应途径显着相关,包括氧化磷酸化和丙氨酸,天冬氨酸,和谷氨酸代谢。然后,我们确定了40个候选基因(例如,HACD2,Cp1,CYP1A2和GPX1)通过STEM和WGCNA分析对饥饿应激的反应。RT-qPCR验证了高通量测序结果的准确性和可靠性。这项研究提供了有关蜗牛越冬存活以及蜗牛适应饥饿胁迫的潜在调节机制的见解。
    Cipangopaludina chinensis, as a financially significant species in China, represents a gastropod in nature which frequently encounters starvation stress owing to its limited prey options. However, the underlying response mechanisms to combat starvation have not been investigated in depth. We collected C. chinensis under several times of starvation stress (0, 7, 30, and 60 days) for nutrient, biochemical characteristics and transcriptome analyses. The results showed that prolonged starvation stress (> 30 days) caused obvious fluctuations in the nutrient composition of snails, with dramatic reductions in body weight, survival and digestive enzyme activity (amylase, protease, and lipase), and markedly enhanced the antioxidant enzyme activities of the snails. Comparative transcriptome analyses revealed 3538 differentially expressed genes (DEGs), which were significantly associated with specific starvation stress-responsive pathways, including oxidative phosphorylation and alanine, aspartate, and glutamate metabolism. Then, we identified 40 candidate genes (e.g., HACD2, Cp1, CYP1A2, and GPX1) response to starvation stress through STEM and WGCNA analyses. RT-qPCR verified the accuracy and reliability of the high-throughput sequencing results. This study provides insights into snail overwintering survival and the potential regulatory mechanisms of snail adaptation to starvation stress.
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  • 文章类型: Journal Article
    在这项研究中,我们调查了有和没有COVID-19的精神分裂症患者的临床生化值和线粒体质量的差异,从而为COVID-19阳性精神分裂症患者的治疗和管理提供帮助.
    我们进行了探索,12月患者数据的回顾性回顾6,2022,1月2023年31日。在此期间,共确定了1696名精神病住院患者(921名精神分裂症患者和775名被诊断为其他精神疾病)。最后,60名精神分裂症患者被纳入我们的研究,其中20例感染了冠状病毒2综合征(SARS-CoV-2)。分析所有精神分裂症患者的血清生化水平和T淋巴细胞的单细胞线粒体质量(SCMM)。
    血清谷草转氨酶(AST)水平,碱性磷酸酶(ALP),COVID-19(SCZ-C)组精神分裂症患者的肌酐(Cr)和乳酸脱氢酶(LDH)明显升高。此外,SCZ-C组显示较低的CD3+,SCZ组T淋巴细胞的CD3+CD4+和CD3+CD8+细胞计数较SCMM高。此外,在阳性和阴性综合征量表(PANSS)上,T细胞亚群计数与阳性症状评分呈正相关。
    我们的研究结果表明,患有COVID-19的精神分裂症患者具有T淋巴细胞线粒体损伤的表型和较高的血清AST水平,ALP,Cr和LDH,这可能为在随后的传染病传播期间治疗精神分裂症患者提供证据。
    UNASSIGNED: In this study, we investigated the differences in clinical biochemical values and mitochondrial mass between schizophrenia patients with and without COVID-19, so as to provide assistance to the treatment and management of COVID-19 positive patients with schizophrenia.
    UNASSIGNED: We undertook an exploratory, retrospective review of patient data from Dec. 6, 2022, to Jan. 31, 2023. A total of 1696 inpatients with psychosis (921 schizophrenia patients and 775 diagnosed with other mental diseases) during this period were identified. Finally, 60 schizophrenia patients were enrolled in our study, and 20 of them were infected with syndrome coronavirus 2 (SARS-CoV-2). The serum biochemical levels and single-cell mitochondrial mass (SCMM) of the T lymphocytes of all schizophrenia patients were analyzed.
    UNASSIGNED: The serum levels of aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatinine (Cr) and lactate dehydrogenase (LDH) were significantly higher in schizophrenia patients with COVID-19 (SCZ-C) group. In addition, the SCZ-C group showed lower CD3+, CD3+CD4+ and CD3+CD8+ cell counts and higher SCMM of T lymphocytes compared to SCZ group. Furthermore, positive correlations were found between the T-cell subpopulation counts and positive symptom scores on the Positive and Negative Syndrome Scale (PANSS).
    UNASSIGNED: Our study findings showed that schizophrenia patients with COVID-19 have a phenotype of mitochondrial damage in T lymphocytes and higher serum levels of AST, ALP, Cr and LDH, which might provide evidence for treating individuals with schizophrenia during subsequent spread of infectious disease.
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  • 文章类型: Journal Article
    多枝苔藓种的两种种质,即德国和塞尔维亚基因型,受到盐胁迫,旨在研究该物种的发育和生理特征。将各种浓度的氯化钠应用于两种苔藓基因型的无菌体外培养物中,并跟踪生长参数和生理特征变化。根据形态发育参数和生存指数推断,与德国基因型相比,塞尔维亚基因型对盐胁迫的抵抗力更高。然而,两种苔藓基因型均在最高浓度(500mM)下存活。不出所料,短时间接触盐很容易克服。在胁迫期间没有观察到糖含量和变化的清晰模式,但它们肯定包括在Formosum的盐应激反应和耐受性中。更长的胁迫增加了两种基因型的总叶绿素含量。在短期施加的盐胁迫中,塞尔维亚基因型具有较高的总叶绿素浓度来控制未受胁迫的植物,而德国基因型降低了叶绿素的总量。同样,类胡萝卜素在塞尔维亚基因型中显示明显更高,在无应力和经过处理的植物中,与德国相比。与德国登录名相比,塞尔维亚基因型的生育酚含量更高,在受控的无胁迫和随后的短期和长期胁迫的小植株中。总的来说,我们可以假设Formosum对盐胁迫具有出乎意料的耐受性,并且在整个欧洲人口的各种种质中存在差异,由两个随机选择的基因型,这很可能是不同遗传结构的结果。
    The two accessions of the polytrichaceous moss species Polytrichum formosum, namely German and Serbian genotypes, were subjected to salt stress, aiming to study the species\' developmental and physiological features. Various concentrations of sodium chloride were applied to an axenic in vitro culture of the two moss genotypes, and the growth parameters as well as physiological feature changes were followed. As inferred by the morpho-developmental parameters and survival index, the Serbian genotype showed higher resistance to salt stress as compared to the German one. However, both moss genotypes survived the highest applied concentration (500 mM). As expected, short exposures to salt were rather easily overcome. No clear patterns in sugar content and changes were observed during the stress, but they are surely included in salt stress response and tolerance in P. formosum. Longer stress increased total chlorophyll content in both genotypes. In short-term applied salt stress, the Serbian genotype had a higher total chlorophyll concentration to control unstressed plants, while the German genotype decreased the total amount of chlorophyll. Similarly, carotenoids were shown to be significantly higher in the Serbian genotype, both in unstressed and treated plants, compared to the German one. The contents of tocopherols were higher in the Serbian genotype in controlled unstressed and subsequently short- and long-stressed plantlets compared to the German accession. In general, we can assume that P. formosum is unexpectedly tolerant to salt stress and that there are differences within various accessions of overall European populations, as referred by two randomly selected genotypes, which is most probably a consequence of different genetic structure.
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  • 文章类型: Journal Article
    自噬是一种以双膜自噬体形成为特征的细胞分解代谢过程。透射电子显微镜是最严格的方法,可以清楚地看到自噬吞噬和降解。大量研究表明自噬与消化密切相关,分泌,和胃肠(GI)细胞的再生。然而,自噬在胃肠道疾病中的作用仍存在争议。本文重点介绍自噬在胃肠道疾病中的形态学和生化特征,为其诊断和治疗提供新的思路。
    Autophagy is a cellular catabolic process characterized by the formation of double-membrane autophagosomes. Transmission electron microscopy is the most rigorous method to clearly visualize autophagic engulfment and degradation. A large number of studies have shown that autophagy is closely related to the digestion, secretion, and regeneration of gastrointestinal (GI) cells. However, the role of autophagy in GI diseases remains controversial. This article focuses on the morphological and biochemical characteristics of autophagy in GI diseases, in order to provide new ideas for their diagnosis and treatment.
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  • 文章类型: Journal Article
    近几十年来,硫酸化的岩藻因其不同的生物活性而引起了广泛的研究兴趣。岩藻酶是研究硫酸化岩藻聚糖的重要工具。本研究报道了三种GH174家族内切-1,3-岩藻聚糖酶的生物信息学分析和生化特性。其中,Fun174Rm和Fun174Sb在报道的岩藻酶中显示出最高的最佳反应温度,Fun174Sb具有良好的热稳定性和催化效率。Fun174Rm显示了一种随机的内在行为方式,而Fun174Ri和Fun174Sb以持续的方式水解硫酸化的岩藻聚糖。UPLC-MS和NMR分析证实,这三种酶催化了来自异形皮的硫酸化岩藻聚糖中Fucp2S和Fucp2S之间的α(1→3)-键的裂解。这些酶证明了新的切割特异性,可以接受位于-1和+1亚位的α-Fucp2S残基。这些生物技术工具的获得将有利于硫酸岩藻聚糖的深入研究。
    Sulfated fucans have garnered extensive research interest in recent decades due to their varied bioactivity. Fucanases are important tools for investigating sulfated fucans. This study reported the bioinformatic analysis and biochemical properties of three GH174 family endo-1,3-fucanases. Wherein, Fun174Rm and Fun174Sb showed the highest optimal reaction temperature among the reported fucanases, and Fun174Sb possessed favorable thermostability and catalysis efficiency. Fun174Rm displayed a random endo-acting manner, while Fun174Ri and Fun174Sb hydrolyzed sulfated fucan in processive manners. UPLC-MS and NMR analyses confirmed that the three enzymes catalyze cleavage of the α(1 → 3)-bonds between Fucp2S and Fucp2S in the sulfated fucan from Isostichopus badionotus. These enzymes demonstrated novel cleavage specificities, which could accept α-Fucp2S residues at subsites -1 and + 1. The acquiring of these biotechnological tools would be beneficial to the in-depth research of sulfated fucans.
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  • 文章类型: Journal Article
    一种来自黄杆菌属的新型淀粉酶AmyFlA。NAU1659,AmyFLA,克隆并在大肠杆菌中表达。基于系统发育和功能分析,它被鉴定为GH13_46亚家族的新成员,共享高序列同一性。该蛋白质预计由620个氨基酸组成,具有25个氨基酸的推定信号肽。该酶能够在50°C下在50mM磷酸盐缓冲液(pH6.0)中以352.97U/mg的比活性水解可溶性淀粉。在最佳条件下,AmyFLA的Km和Vmax值分别为3.15mg/ml和566.36µmol·ml-1·min-1。在存在1mMCa2+的情况下,其对淀粉的活性提高了63%,表明AmyFLA是Ca2+依赖性淀粉酶。与报道的产麦芽糖淀粉酶相比,AmyFlA在反应开始时产生较低种类的中间寡糖,使得产物混合物含有较高比例的麦芽糖。这些结果表明AmyFLA可能在高麦芽糖糖浆生产中具有潜在的应用价值。
    A novel amylase AmyFlA from Flavobacterium sp. NAU1659, AmyFlA, was cloned and expressed in Esherichia coli. Based on phylogenetic and functional analysis, it was identified as a novel member of the subfamily GH13_46, sharing high sequence identity. The protein was predicted to consist of 620 amino acids, with a putative signal peptide of 25 amino acids. The enzyme was able to hydrolyze soluble starch with a specific activity of 352.97 U/mg at 50 °C in 50 mM phosphate buffer (pH 6.0). The Km and Vmax values of AmyFlA were respectively 3.15 mg/ml and 566.36 µmol·ml-1·min-1 under optimal conditions. Its activity towards starch was enhanced by 63% in the presence of 1 mM Ca2+, indicating that AmyFlA was a Ca2+-dependent amylase. Compared to the reported maltogenic amylases, AmyFlA produced a lower variety of intermediate oligosaccharides at the start of the reaction so that the product mixture contained a higher proportion of maltose. These results indicate that AmyFlA may be potential application value in the production of high-maltose syrup.
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  • 文章类型: Journal Article
    嗜冷黄杆菌是沙门氏菌和虹鳟鱼鱼苗综合征中细菌性冷水病的病原体。这种病原体已在全球范围内存在,并且可以水平和垂直传播。然而,2018年9月之前在韩国没有记录.在这项研究中,目的是对从患病的虹鳟鱼中分离出的嗜冷黄杆菌菌株FPRT1进行基因型和表型鉴定。我们还进行了各种调查,以更好地了解其影响并评估潜在的控制措施。我们获得了50条虹鳟鱼(重量约70克),并将其转移到实验室水族馆。在最初的适应期,我们观察了死亡率,并检查了受影响的鱼类的临床体征。我们使用添加葡萄糖的胰蛋白胨酵母提取物盐琼脂从感染的虹鳟鱼的脾脏中分离出细菌,命名此FPRT1。进行了抗生素敏感性测试,从结果来看,我们选择恩诺沙星对鳟鱼口服给药,以降低死亡率.为了评估致病性,我们在不同的水温(8、15和22°C)下将鳟鱼暴露于FPRT1。进行基因组分析以鉴定FPRT1与欧洲菌株的血清型和相关性。受影响的鱼类表现出临床症状,例如下颌骨的溃疡性病变,贫血与苍白的ill,眼球突出症,粘液分泌增加。内部症状包括肝脏苍白和脾脏肿大。FPRT1对红霉素敏感,恩诺沙星,氟苯尼考,土霉素,还有庆大霉素,但对草酸和磺胺甲恶唑/甲氧苄啶有抗性。口服恩诺沙星导致死亡率从28%降低到6%。致病性测试显示,由于FPRT1在不同温度下的死亡率不同。腹膜内和肌肉注射在8°C时观察到最高的比率(范围从43%到100%),和较低的比率发生在22°C(范围从0%到30%),肌肉注射显示更高的敏感性。基因组分析确定FPRT1为血清型2,并表明其与欧洲菌株的遗传关系密切,基于核心基因组和非必需基因组。我们的菌株与欧洲菌株之间的基因组相似性表明,细菌可能通过从欧洲进口受精卵而传播。总之,这项研究强调了以前未记录的病原体的引入(F.嗜冷菌)进入韩国虹鳟鱼种群。这种病原体的检测及其致病性评估不仅对于了解其对当地水产养殖的影响很重要,而且对于建立监测和控制措施以防止该地区的进一步传播和爆发也很重要。
    Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease in salmonids and rainbow trout fry syndrome. This pathogen has attained a global presence and can spread both horizontally and vertically. However, it was not documented in Korea before September 2018. In this study, the objectives were to characterize Flavobacterium psychrophilum strain FPRT1, isolated from diseased rainbow trout genotypically and phenotypically. We also conducted various investigations to better understand its impact and assess potential control measures. We acquired fifty rainbow trout (approximately 70 g in weight) and transferred them to a laboratory aquarium. During the initial acclimation period, we observed mortality and examined affected fish for clinical signs. We isolated the bacterium from the spleen of infected rainbow trout using tryptone yeast extract salts agar supplemented with glucose, naming this FPRT1. Antibiotic susceptibility testing was carried out, and from the result, we selected enrofloxacin to administer to the trout orally to reduce mortality. To evaluate pathogenicity, we exposed the trout to FPRT1 at different water temperatures (8, 15, and 22 °C). Genomic analysis was conducted to identify the serotype and relatedness of FPRT1 to European strains. Affected fish displayed clinical signs, such as ulcerative lesions in the mandible, anemia with pale gills, exophthalmia, and increased mucus secretion. Internal symptoms included pale liver and enlarged spleen. FPRT1 was susceptible to erythromycin, enrofloxacin, florfenicol, oxytetracycline, and gentamicin, but resistant to oxolinic acid and sulfamethoxazole/trimethoprim. Oral administration of enrofloxacin resulted in a decrease in mortality from 28% to 6%. Pathogenicity tests revealed varying mortality rates due to FPRT1 at different temperatures. The highest rates were observed at 8 °C (ranging from 43% to 100%) for both intraperitoneal and intramuscular injections, and lower rates occurred at 22 °C (ranging from 0% to 30%), with intramuscular injections displaying higher susceptibility. Genomic analysis identified FPRT1 as serotype 2 and indicated its close genetic relationship with European strains based on the core genome and dispensable genome. The substantial genomic similarity between our strain and European strains suggests the possibility of bacterial spread through the importation of fertilized eggs from Europe. In conclusion, this study highlights the introduction of the previously undocumented pathogen (F. psychrophilum) into Korean rainbow trout populations. The detection of this pathogen and its pathogenicity assessment is not only important for understanding its impact on local aquaculture but also for establishing surveillance and control measures to prevent further transmission and outbreaks in the region.
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  • 文章类型: Journal Article
    Alagille综合征(ALGS)是一种多系统疾病,其特征是胆汁淤积和胆管缺乏肝活检和心脏的可变受累,骷髅,眼睛,肾脏,并由JAG1或NOTCH2基因的致病变异引起。临床表型的可变表达和缺乏基因型-表型相关性导致显著的诊断困难。在这里,我们对18例诊断为ALGS的胆汁淤积患者进行了分析。我们使用了针对52个基因的编码外显子的NGS面板,包括JAG1和NOTCH2基因。使用Sanger测序来验证受影响的个体和家庭成员中的突变。在16/18(88.9%)中可见特定的面部表型。8/18(44.4%)患者出现心脏缺损(7/8)。5/14(35.7%)患者可见蝶形椎骨。2/18(11.1%)例发现肾脏受累-1例患者有肾囊肿,其中一人患有阻塞性肾积水.对12名儿童进行了眼科检查,只有1人患有后胚毒素(8.3%)。9例进行了经皮肝活检。6/9例(66.7%)发现胆管缺乏。两名患者因肝硬化需要肝移植。我们在JAG1基因中鉴定了9个新变体-8个移码变体(c.1619_1622dupGCTA(p。Tyr541X),c.1160delG(p.Gly387fs),c.964dupT(p。C322fs),c.120delG(p.L40fs),c.1984dupG(p。Ala662Glyfs),c.3168_3169delAG(p。R1056Sfs*51),c.2688delG(p.896CysfsTer49),c.164dupG(p。Cys55fs))和一个错义变体,c.2806T>G(p。Cys936Gly).没有患者出现NOTCH2变异。根据经典标准,在没有基因分析的情况下,我们队列中只有6例患者符合诊断标准.基因检测在ALGS的诊断中很重要,可以帮助将其与其他类型的胆汁淤积区分开。
    Alagille syndrome (ALGS) is a multisystem condition characterized by cholestasis and bile duct paucity on liver biopsy and variable involvement of the heart, skeleton, eyes, kidneys, and face and caused by pathogenic variants in the JAG1 or NOTCH2 gene. The variable expressivity of the clinical phenotype and the lack of genotype-phenotype correlations lead to significant diagnostic difficulties. Here we present an analysis of 18 patients with cholestasis who were diagnosed with ALGS. We used an NGS panel targeting coding exons of 52 genes, including the JAG1 and NOTCH2 genes. Sanger sequencing was used to verify the mutation in the affected individuals and family members. The specific facial phenotype was seen in 16/18 (88.9%). Heart defects were seen in 8/18 (44.4%) patients (pulmonary stenosis in 7/8). Butterfly vertebrae were seen in 5/14 (35.7%) patients. Renal involvement was detected in 2/18 (11.1%) cases-one patient had renal cysts, and one had obstructive hydronephrosis. An ophthalmology examination was performed on 12 children, and only one had posterior embryotoxon (8.3%). A percutaneous liver biopsy was performed in nine cases. Bile duct paucity was detected in six/nine cases (66.7%). Two patients required liver transplantation because of cirrhosis. We identified nine novel variants in the JAG1 gene-eight frameshift variants (c.1619_1622dupGCTA (p.Tyr541X), c.1160delG (p.Gly387fs), c.964dupT (p.C322fs), c.120delG (p.L40fs), c.1984dupG (p.Ala662Glyfs), c.3168_3169delAG (p.R1056Sfs*51), c.2688delG (p.896CysfsTer49), c.164dupG (p.Cys55fs)) and one missense variant, c.2806T > G (p.Cys936Gly). None of the patients presented with NOTCH2 variants. In accordance with the classical criteria, only six patients could meet the diagnostic criteria in our cohort without genetic analysis. Genetic testing is important in the diagnosis of ALGS and can help differentiate it from other types of cholestasis.
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  • 文章类型: Journal Article
    背景:在这项研究中,α-角叉菜酶基因,Car1293是从微球藻的基因组中获得的。从大型藻类表面分离的YNDZ01,印度尼西亚。到目前为止,关于1-角叉菜胶酶和1-角叉菜胶寡糖(CGOS)的抗炎活性的研究很少。为了增强我们对1-角叉菜酶和1-角叉菜胶寡糖的看法,基因序列,蛋白质结构,酶学性质,研究了酶消化产物及其抗炎活性。
    结果:Car1293的基因长度为2,589bp,编码862个氨基酸的酶,与以前报道的任何1-角叉菜酶有34%的相似性。Car1293的空间结构由许多α-螺旋组成,末端有一个β-折叠结合模块,根据与CGOS-DP4配体对接的结果,在结合模块中发现了8个结合位点。重组Car1293对1-角叉菜胶的活性的最适温度和pH分别为50°C和6.0。Car1293的水解产物主要是聚合度(DP)8,次要产物具有DP2、DP4和DP6。酶促水解产物CGOS-DP8在脂多糖诱导的RAW264.7巨噬细胞中显示出比阳性对照L-单甲基精氨酸更大的显著抗炎活性;它抑制NO产生,并显着抑制肿瘤坏死因子-α和白细胞介素-6的分泌。
    结论:由Car1293编码的1-角叉菜胶序列是新的,可以将角叉菜胶水解为具有显著抗炎作用的CGOS-DP8。本研究填补了1-角叉菜胶中寡糖生物活性研究的空白,为天然抗炎剂的开发提供了有希望的数据。本文受版权保护。保留所有权利。
    BACKGROUND: In the present study, the ι-carrageenase gene, Car1293, was obtained from the genome of Microbulbifer sp. YNDZ01, which was isolated from the surface of macroalgae. To date, there are few studies on ι-carrageenase and the anti-inflammatory activity of ι-carrageenan oligosaccharides (CGOS). To enhance our perspective on ι-carrageenase and ι-carrageen oligosaccharides, the sequence, protein structure, enzymatic properties, enzymatic digestion products and anti-inflammatory activity of the gene were investigated.
    RESULTS: The gene length of Car1293 is 2,589 bp, encoding an enzyme with 862 amino acids, which shares 34% similarity with any previously reported ι-carrageenase. The spatial structure of Car1293 consists of many α-helices with a β-fold binding module located at its terminus, and eight binding sites were found in the binding module as a result of docking with CGOS-DP4 ligand. The optimum temperature and pH for the activity of recombinant Car1293 toward ι-carrageenan were 50 °C and 6.0, respectively. The hydrolysates of Car1293 are mainly degree of polymerization (DP)8, with minor products showing DP2, DP4, and DP6. The enzymatic hydrolysates CGOS-DP8 showed prominent anti-inflammatory activity, which was greater than that of the positive control l-monomethylarginine in lipopolysaccharide-induced RAW264.7 macrophages. It inhibited nitric oxide production, as well as significantly inhibited tumor necrosis factor-α and interleukin-6 secretion.
    CONCLUSIONS: The ι-carrageenase sequence encoded by Car1293 is novel and can hydrolyze carrageenan into CGOS-DP8 that has a significant anti-inflammatory effect. The present study fills a gap in the research on the biological activity of oligosaccharides in ι-carrageenan and provides promising data for the development of natural anti-inflammatory agent. © 2023 Society of Chemical Industry.
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