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Chemical Composition and Antibacterial and Antioxidant Activities of Stem Bark Essential Oil and Extracts of Solanecio gigas. 龙舌兰茎皮精油及提取物的化学成分及抑菌抗氧化活性研究
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-07-09 eCollection Date: 2022-01-01 DOI: 10.1155/2022/4900917
Mequanint Molla Yitayeh, Amanu Monie Wassihun

Herbal medication developed from natural resources has to have antibacterial and antioxidant effects. The aim of this research is to look at the chemical makeup of Solanecio gigas (S. gigas) stem bark essential oil (EO), as well as the effectiveness of EO and extracts (chloroform, ethyl acetate, and methanol) against human pathogenic bacteria and their antioxidant activity. The GC-MS analysis identified 23 components, accounting for 98.7% of the total oil containing Methylene chloride (49.2%), sabinene (10.5%), 1-nonene (11.3%), Terpinen-4-ol (6.9%), Camphene (4.3%), γ-terpinene (3.6%), α-phellandrene (2.9%) β-myrcene (2.6%), 1,2,5-Oxadiazol-3-carboxamide, 4,4'-azobis-2,2'-dioxide (2.4%), α-terpinene (1.9%), 1-Octanamine, N-methyl- (1.9%), ρ-cymene (1.6%) as major components. The antibacterial efficacy of the EO and extracts (25, 50, 100, and 200 mg/ml) was demonstrated by the inhibitory zones (8.5 ± 0.47-23.3 ± 0.36 and 7.2 ± 0.25-22.0 ± 0.45 mm), respectively. The MIC values of the extracts and the EO were 120-150 and 240 to <1100 μg/ml, respectively. The EO also demonstrated a significant antibacterial impact. The EO and methanolic extract had free radical scavenging activities with IC50 value, 13.8 ± 0.48 and 4.2 ± 0.04 μg/ml, respectively. In comparison to the other extracts, the methanolic extract had the greatest phenolics (100.2 ± 0.13 μg GAE/mg of dry extract) and flavonoid contents (112.1 ± 0.18 μg CE/mg of dry extract).

从自然资源中开发的草药必须具有抗菌和抗氧化作用。本研究的目的是研究茄干皮精油(EO)的化学组成,以及EO及其提取物(氯仿、乙酸乙酯和甲醇)对人类致病菌的有效性及其抗氧化活性。GC-MS分析鉴定出23种成分,占总油的98.7%,主要成分为二氯甲烷(49.2%)、sabinene(10.5%)、1-壬烯(11.3%)、松油烯-4-醇(6.9%)、Camphene(4.3%)、γ-松油烯(3.6%)、α-茶树烯(2.9%)、β-月桂烯(2.6%)、1,2,5- oxadiazol -3-carboxamide、4,4′-偶氮-2,2′-dioxide(2.4%)、α-松油烯(1.9%)、1-辛胺、n-甲基-(1.9%)、ρ-cymene(1.6%)。EO和提取物(25、50、100和200 mg/ml)的抑菌区分别为(8.5±0.47 ~ 23.3±0.36 mm)和(7.2±0.25 ~ 22.0±0.45 mm)。提取物的MIC值为120 ~ 150 μg/ml, EO值为240 μg/ml。EO也显示出显著的抗菌作用。EO和甲醇提取物的IC50值分别为13.8±0.48和4.2±0.04 μg/ml。与其他提取物相比,甲醇提取物的酚类物质(100.2±0.13 μg CE/mg)和类黄酮含量(112.1±0.18 μg CE/mg)最高。
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引用次数: 8
The Effect of Supplementing Mushroom Growing Substrates on the Bioactive Compounds, Antimicrobial Activity, and Antioxidant Activity of Pleurotus ostreatus. 添加蘑菇生长基质对平菇生物活性物质、抑菌活性和抗氧化活性的影响。
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-06-27 eCollection Date: 2022-01-01 DOI: 10.1155/2022/9436614
Senzosenkosi Surprise Mkhize, Mthokozisi Blessing Cedric Simelane, Ishmael Nkoana Mongalo, Ofentse Jacob Pooe

Pleurotus ostreatus mushroom contains important bioactive compounds and has several biological activities; however, mushroom growing substrates have major influence on chemical and functional characteristics of the mushroom. Hence, the study aimed to evaluate the influence of supplementing mushroom growing substrates with wheat bran (WB) towards yield/productivity, bioactive compounds, and antimicrobial and antioxidant activity of P. ostreatus. The mushroom was cultivated on sugarcane substrates supplemented with increasing levels of WB (0%-20%). The mushroom extracts were screened for bioactive compounds using gas chromatography-mass spectrometry (GC-MS). Antimicrobial activity was carried out using microplate assay, while antioxidant potential was investigated using reducing power assay. The addition of supplements on mushroom growing substrates had an influence on mushroom yield; hence, higher supplementation (18% and 20%) produced higher yield. The GC-MS revealed several bioactive compounds with known activity, such as vitamin E, phenol, fatty acids, and terpenoids. Concentration-dependent antioxidant activity was observed; hence, extracts at higher concentrations gave significantly higher reducing power. The P. ostreatus extract had antimicrobial activity against all the tested organisms, with S. aureus showing high susceptibility to most of the extracts. However, mushrooms grown on bagasse substrates supplemented with 14% (0.02 mg/ml) and 20% WB (0.08 mg/ml) proved to have better antimicrobial activity on Escherichia coli. The difference in susceptibility demonstrates that substrates type and composition could have an influence on bioactive compounds found within mushrooms, also influencing medicinal properties of edible mushroom. Thus, supplementing mushroom growing substrates not only improve yield, but also can contribute to bioactive compounds with medicinal potential.

平菇含有重要的生物活性化合物,具有多种生物活性;然而,蘑菇生长基质对蘑菇的化学和功能特性有重要影响。因此,本研究旨在评价在蘑菇生长基质中添加麦麸(WB)对P. ostreatus的产量/生产力、生物活性物质以及抗菌和抗氧化活性的影响。在添加WB(0% ~ 20%)的甘蔗基质上培养。采用气相色谱-质谱联用技术对香菇提取物进行生物活性成分筛选。用微孔板法测定其抑菌活性,用还原力法测定其抗氧化能力。在蘑菇生长基质上添加补剂对蘑菇产量有影响;因此,添加量越高(18%和20%)产量越高。GC-MS显示了几种已知活性的生物活性化合物,如维生素E、酚、脂肪酸和萜类化合物。观察浓度依赖性抗氧化活性;因此,浓度越高的提取物还原力越强。该提取物对所有被试菌均有抑菌活性,其中金黄色葡萄球菌对大部分提取物均有较高的敏感性。结果表明,在添加14% (0.02 mg/ml)和20% WB (0.08 mg/ml)的甘蔗渣基质上生长的蘑菇对大肠杆菌具有较好的抑菌活性。这种敏感性的差异表明,底物类型和组成可能会影响蘑菇中发现的生物活性物质,也会影响食用菌的药用特性。因此,补充菌类生长基质不仅可以提高产量,而且可以促进具有药用潜力的生物活性化合物的产生。
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引用次数: 4
Screening of Aqueous Extract of Persea americana Seeds for Alpha-Glucosidase Inhibitors 美洲Persea americana种子水提取物对α-葡萄糖苷酶抑制剂的筛选
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-05-14 DOI: 10.1155/2022/3492203
T. A. Lawal
Activity of α-glucosidase enzyme in the gastrointestinal tract has been implicated in postprandial hyperglycaemia. If not properly controlled, postprandial hyperglycaemia might progress to diabetes mellitus, a metabolic syndrome. Diabetes is associated with many complications such as retinopathy, heart attack, nephropathy, neuropathy, stroke, and lower limb amputation. Antidiabetic medications presently in use have little effect on postprandial glycaemic excursion and hence do not bring down the blood glucose level to baseline. This study extracted, fractionated, and screened the aqueous extract of Persea americana seeds for hypoglycaemic potential. Inhibitory effects of the fractions and subfractions of the extract on α-glucosidase activity were investigated. The most active subfraction was subjected to Fourier transform infrared (FTIR) and gas chromatography mass spectroscopy (GC-MS) analysis to elucidate the active components. The active subfraction showed a significant inhibition (p < 0.05) on α-glucosidase. The subfraction competitively inhibits α-glucosidase (with IC50 = 09.48 ± 0.58 μg/mL), though less potent than the standard drug, acarbose (IC50 = 06.45 ± 0.47 μg/mL). FTIR analysis of the subfraction showed the presence of carbonyl group, hydroxy group, carboxyl group, double bonds, methylene, and methyl groups. GC-MS analysis suggests the presence of cis-11,14-eicosadienoic acid, catechin, and chlorogenic acid as the active components. In conclusion, the components obtained from this study can be synthesised in the laboratory to further confirm their hypoglycaemic activity. The most active subfraction can be explored further to confirm its inhibitory activity against the enzyme and to determine its extent in the treatment of diabetes mellitus in vivo.
胃肠道中α-葡萄糖苷酶的活性与餐后高血糖有关。如果控制不当,餐后高血糖可能发展为糖尿病,一种代谢综合征。糖尿病与许多并发症有关,如视网膜病变、心脏病发作、肾病、神经病变、中风和下肢截肢。目前使用的抗糖尿病药物对餐后血糖漂移几乎没有影响,因此不会将血糖水平降至基线。本研究提取、分级并筛选了美洲Persea americana种子的水提取物的低血糖潜力。研究了提取物的级分和亚级分对α-葡萄糖苷酶活性的抑制作用。对最具活性的亚组分进行傅立叶变换红外光谱(FTIR)和气相色谱-质谱(GC-MS)分析,以阐明活性成分。活性亚组分对α-葡萄糖苷酶有显著的抑制作用(p<0.05)。该亚组分竞争性抑制α-葡萄糖苷酶(IC50 = 9.48 ± 0.58 μg/mL),尽管药效低于标准药物阿卡波糖(IC50 = 6.45 ± 0.47 μg/mL)。亚组分的FTIR分析显示存在羰基、羟基、羧基、双键、亚甲基和甲基。GC-MS分析表明存在顺式-11,14-二十碳二烯酸、儿茶素和绿原酸作为活性成分。总之,从这项研究中获得的成分可以在实验室中合成,以进一步证实其降血糖活性。可以进一步探索最活性的亚组分,以确认其对酶的抑制活性,并确定其在体内治疗糖尿病中的程度。
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引用次数: 0
miR-21 Regulates Immune Balance Mediated by Th17/Treg in Peripheral Blood of Septic Rats during the Early Phase through Apoptosis Pathway miR-21通过细胞凋亡途径调节脓毒症大鼠早期外周血Th17/Treg介导的免疫平衡
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-04-27 DOI: 10.1155/2022/9948229
Cheng Liu, Qi Zou
Objective To study the mechanism by which miR-21 regulates the differentiation and function of Th17/Treg cells in sepsis. Methods A rat model with sepsis was made by cecal ligation and puncture (CLP). Then, some of the septic rats were transfected with miR-21 mimic or inhibitor by liposome. At 48 hours, lymphocytes and plasma from septic rats were isolated for further experimental detection. The expression of miR-21 in lymphocytes was detected by Polymerase Chain Reaction (PCR); the differentiation of Th17/Treg cells was counted by flow cytometry; lymphocyte apoptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. The caspase-3/9 proteins were tested by Western blot; IL-10 and IL-17 were detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with the sepsis group (SP group), the Th17 cells increased significantly, the Treg cells decreased significantly, the apoptosis rate of lymphocytes decreased significantly, the mRNA and proteins of caspase-3/9 decreased significantly, the IL-17 decreased, and the IL-10 increased in the sepsis group transfected with miR-21 (SP + miR-21 mimic group). After transfection of miR-21 inhibitor, the results were almost opposite to those of SP + miR-21 mimic group. Conclusions The differentiation and function of Th17/Treg cells were regulated by miR-21 in sepsis through caspase pathway.
目的探讨miR-21在脓毒症中调节Th17/Treg细胞分化和功能的机制。方法采用盲肠结扎穿刺法建立大鼠败血症模型。然后,通过脂质体用miR-21模拟物或抑制剂转染一些脓毒症大鼠。48 小时后,分离脓毒症大鼠的淋巴细胞和血浆进行进一步的实验检测。用聚合酶链式反应(PCR)检测淋巴细胞中miR-21的表达;通过流式细胞术计数Th17/Treg细胞的分化;用末端脱氧核苷酸转移酶介导的dUTP生物素缺口末端标记法(TUNEL)观察淋巴细胞凋亡。用蛋白质印迹法检测胱天蛋白酶3/9蛋白;用酶联免疫吸附试验(ELISA)检测IL-10和IL-17。结果与败血症组(SP组)相比,转染miR-21的败血症组Th17细胞显著增加,Treg细胞显著减少,淋巴细胞凋亡率显著降低,半胱氨酸天冬氨酸蛋白酶3/9 mRNA和蛋白显著降低,IL-17降低,IL-10升高 + miR-21模拟组)。转染miR-21抑制剂后,结果几乎与SP相反 + miR-21模拟组。结论miR-21通过caspase途径调控脓毒症Th17/Treg细胞的分化和功能。
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引用次数: 0
Updates on Prevalence and Trend Status of Visceral Leishmaniasis at Two Health Facilities in Amhara Regional State, Northwest Ethiopia: A Retrospective Study 埃塞俄比亚西北部阿姆哈拉州两家卫生机构内脏利什曼病流行和趋势状况的最新情况:一项回顾性研究
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-04-15 DOI: 10.1155/2022/3603892
Mulat Yimer, E. Nibret, Gizachew Yismaw
Ethiopia is one of the countries accounted for over 90% of annual visceral leishmaniasis incidence. Despite this, yet there are no active and passive surveillance activities in the Amhara Region that will give up-to-date information about the disease status at the health facility levels. Therefore, this study aimed to report up-to-date information about visceral leishmaniasis and its trend status at two health facilities and the surrounding areas. A retrospective study from October 2017 to May 2021 was conducted by reviewing patient records at Metema and Addis Zemen Hospitals. Data on Sex, age, occupation, residence, month, year, and rK39 test results were collected using a questionnaire and were analyzed using Statistical Package for Social Sciences (SPSS) version 20. The chi-square test was used to see the association between variables. p < 0.05 was considered as statistically significant. Of the 2,703 visceral leishmaniasis suspected cases diagnosed with the rK39 test, 877 (32.4%) were confirmed (positive) cases. Monthly and yearly trends depicted that the largest number of suspected cases was reported in October and 2018, respectively. Daily laborers were the most affected individuals in Metema areas.
埃塞俄比亚是年内脏利什曼病发病率超过90%的国家之一。尽管如此,阿姆哈拉地区还没有主动和被动的监测活动,可以在卫生设施层面提供有关疾病状况的最新信息。因此,本研究旨在报告两个卫生机构及其周边地区内脏利什曼病的最新信息及其趋势。2017年10月至2021年5月,通过审查Metema和Addis Zemen医院的患者记录进行了一项回顾性研究。使用问卷收集性别、年龄、职业、居住地、月份、年份和rK39测试结果的数据,并使用社会科学统计软件包(SPSS)版本20进行分析。卡方检验用于观察变量之间的关联。p < 0.05被认为具有统计学意义。在通过rK39检测诊断的2703例内脏利什曼病疑似病例中,877例(32.4%)为确诊(阳性)病例。月度和年度趋势显示,10月和2018年报告的疑似病例数量最多。日常工作者是Metema地区受影响最大的人。
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引用次数: 6
Bioassay-Guided Fractionation, ESI-MS Scan, Phytochemical Screening, and Antiplasmodial Activity of Afzelia africana. 非洲梧桐的生物检测导向分离、ESI-MS扫描、植物化学筛选和抗疟原虫活性研究
IF 3.4 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-04-13 eCollection Date: 2022-01-01 DOI: 10.1155/2022/6895560
Bright Yaw Vigbedor, Jonathan Osei-Owusu, Ralph Kwakye, David Neglo

Background: Afzelia africana is a plant species with reported numerous medicinal potentials and secondary metabolites. Various parts of the plant have been applied for the treatment of hernia, rheumatism, pain, lumbago, malaria, etc. The study seeks to evaluate the phytochemical constituents, antiplasmodial, and ESI-MS scan of bioassay-guided fractions from the methanol extract of the bark of the plant.

Aims: The main aim of the study was to carry out bioassay-guided fractionation of the crude methanol extract of Afzelia africana in order to isolate fractions and to evaluate their antiplasmodial activities and ESI-MS fingerprints.

Methods: The methods employed include column chromatographic fractionation, phytochemical screening, antiplasmodial activity (malaria SYBER green assay (MSF)), and ESI-MS profile (full ESI-MS scan).

Results: The column chromatographic fractionation and phytochemical screening of the plant led to the separation of the following four fractions: 1 (flavonoids, phenolics, glycosides, terpenoids, and steroids), 2 (alkaloids, anthraquinones, flavonoids, phenolics, glycosides, terpenoids, and steroids), 3 (anthraquinones, flavonoids, phenolics, glycosides, terpenoids, and steroids), and 4 (alkaloids, flavonoids, phenolics, glycosides, terpenoids, and steroids). The antiplasmodial activities of the fractions were tested against the 3D7 strain of Plasmodium falciparum with reported stronger activities for 1 (IC50: 0.097 ± 0.034 μg/mL) and 3 (IC50: 1.43 ± 0.072 μg/mL), and weaker activities for 2 (IC50: >100 μg/mL) and 4 (IC50: 37.09 ± 6.14 μg/mL). The full ESI-MS fingerprint of fractions 1, 2, 3, and 4 revealed the presence of 14, 24, 34, and 37 major molecular ions or compounds in each fraction, respectively.

背景非洲泽兰是一种具有多种药用潜力和次生代谢产物的植物。该植物的各个部分已被应用于治疗疝气、风湿病、疼痛、腰痛、疟疾等。该研究旨在评估该植物树皮甲醇提取物的植物化学成分、抗疟原虫和ESI-MS扫描的生物测定导向组分。本研究的主要目的是对非洲泽兰粗甲醇提取物进行生物测定引导的分级,以分离部分并评价其抗疟原虫活性和ESI-MS指纹图谱。方法采用柱色谱分离、植物化学筛选、抗疟原虫活性(疟疾SYBER绿色测定法(MSF))和ESI-MS图谱(全ESI-MS扫描)。结果对该植物进行柱色谱分离和植物化学筛选,分离出以下四个组分:1(黄酮类、酚类、糖苷类、萜类和类固醇)、2(生物碱、蒽醌类、黄酮类、苯酚类、糖苷、萜类、类固醇)、3(蒽醌类、类黄酮、酚类,和4(生物碱、类黄酮、酚类、糖苷、萜类和类固醇)。对恶性疟原虫3D7株测试了组分的抗疟原虫活性,据报道其对1的活性更强(IC50:0.097 ± 0.034 μg/mL)和3(IC50:1.43 ± 0.072 μg/mL),2的活性较弱(IC50:>100 μg/mL)和4(IC50:37.09 ± 6.14 μg/mL)。组分1、2、3和4的完整ESI-MS指纹显示,每个组分中分别存在14、24、34和37个主要分子离子或化合物。
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引用次数: 0
Annotating Spike Protein Polymorphic Amino Acids of Variants of SARS-CoV-2, Including Omicron 包括奥密克戎在内的严重急性呼吸系统综合征冠状病毒2型变体的刺突蛋白多态性氨基酸注释
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-04-11 DOI: 10.1155/2022/2164749
G. N. Mahardika, Nyoman B Mahendra, B. K. Mahardika, I. Suardana, M. Pharmawati
The prolonged global spread and community transmission of severe acute respiratory syndrome virus 2 (SARS-CoV-2) has led to the emergence of variants and brought questions regarding disease severity and vaccine effectiveness. We conducted simple bioinformatics on the spike gene of a representative of each variant. The data show that a number of polymorphic amino acids are located mostly on the amino-terminal side of the S1/S2 cleavage site. The Omicron variant diverges from the others, with the highest number of amino acid substitutions, including the receptor-binding site (RBS), epitopes, S1/S2 cleavage site, fusion peptide, and heptad repeat 1. The current sharp global increase in the frequency of the Omicron genome constitutes evidence of its high community transmissibility. In conclusion, the proposed guideline could give an immediate insight of the probable biological nature of any variant of SARS-Cov-2. As the Omicron diverged the farthest from the original pandemic strain, Wuhan-Hu-1, we expect different epidemiological and clinical patterns of Omicron cases. On vaccine efficacy, slight changes in some epitopes while others are conserved should not lead to a significant reduction in the effectiveness of an approved vaccine.
严重急性呼吸综合征2型病毒(严重急性呼吸系统综合征冠状病毒2型)的长期全球传播和社区传播导致了变种的出现,并带来了有关疾病严重性和疫苗有效性的问题。我们对每个变体的代表性刺突基因进行了简单的生物信息学研究。数据显示,许多多态性氨基酸主要位于S1/S2切割位点的氨基末端侧。奥密克戎变体与其他变体不同,具有最高数量的氨基酸取代,包括受体结合位点(RBS)、表位、S1/S2切割位点、融合肽和七肽重复序列1。目前全球奥密克戎基因组频率的急剧上升证明了其高度的社区传播性。总之,拟议的指南可以立即了解严重急性呼吸系统综合征冠状病毒2型任何变体的可能生物学性质。由于奥密克戎与最初的流行毒株武汉胡1变异最远,我们预计奥密克龙病例的流行病学和临床模式会有所不同。在疫苗效力方面,一些表位的微小变化,而另一些表位是保守的,不应导致批准疫苗的效力显著降低。
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引用次数: 1
Retracted: Exploring Drug Targets in Isoprenoid Biosynthetic Pathway for Plasmodium falciparum 撤回:探索恶性疟原虫异丙肾上腺素生物合成途径中的药物靶点
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-03-15 DOI: 10.1155/2022/8426183
Biochemistry Research International
[This retracts the article DOI: 10.1155/2014/657189.].
[本文撤回文章DOI: 10.1155/2014/657189]。
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引用次数: 0
The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid. 几丁质-膨润土杂化固定化提高烟曲霉α-淀粉酶的稳定性。
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-03-14 eCollection Date: 2022-01-01 DOI: 10.1155/2022/5692438
Ezra Rheinsky Tiarsa, Yandri Yandri, Tati Suhartati, Heri Satria, Bambang Irawan, Sutopo Hadi

Enzyme immobilization is a powerful method to improve the stability, reuse, and enzymatic properties of enzymes. The immobilization of the α-amylase enzyme from Aspergillus fumigatus on a chitin-bentonite (CB) hybrid has been studied to improve its stability. Therefore, this study aims to obtain the higher stability of α-amylase enzyme to reduce industrial costs. The procedures were performed as follows: production, isolation, partial purification, immobilization, and characterization of the free and immobilized enzymes. The CB hybrid was synthesized by bentonite, chitin, and glutaraldehyde as a cross-linker. The free enzyme was immobilized onto CB hybrid using 0.1 M phosphate buffer pH 7.5. The free and immobilized enzymes were characterized by optimum temperature, Michaelis constant (K M), maximum velocity (V max), thermal inactivation rate constant (k i ), half-life (t 1/2), and transformation of free energy because of denaturation (ΔG i ). The free enzyme has optimum temperature of 55°C, K M  = 3.04 mg mL-1 substrate, V max=10.90 μmolemL-1min-1, k i  = 0.0171 min-1, t 1/2 = 40.53 min, and ΔG i  = 104.47 kJ mole-1. Meanwhile, the immobilized enzyme has optimum temperature of 60°C, K M  = 11.57 mg mL-1 substrate, V max=3.37 μmolemL-1min-1, k i  = 0.0045 min-1, t 1/2 = 154.00 min, and ΔG i  = 108.17 kJ mole-1. After sixth cycle of reuse, the residual activity of the immobilized enzyme was 38%. The improvement in the stability of α-amylase immobilized on the CB hybrid based on the increase in half-life was four times of the free enzyme.

酶固定化是提高酶的稳定性、可重复利用性和酶学性能的一种有效方法。为提高烟曲霉α-淀粉酶的稳定性,研究了几丁质膨润土(CB)复合材料对烟曲霉α-淀粉酶的固定化作用。因此,本研究旨在获得更高稳定性的α-淀粉酶,以降低工业成本。步骤如下:游离酶和固定化酶的生产、分离、部分纯化、固定化和表征。以膨润土、几丁质和戊二醛为交联剂合成了炭黑杂化物。用0.1 M pH 7.5的磷酸缓冲液将游离酶固定在CB杂交体上。通过最适温度、Michaelis常数(K M)、最大速度(V max)、热失活速率常数(K i)、半衰期(t1 /2)和变性自由能转化(ΔG i)对游离酶和固定化酶进行表征。游离酶的最适温度为55℃,km = 3.04 mg mL-1底物,vmax =10.90 μmolemL-1min-1, ki = 0.0171 min-1, t1 /2 = 40.53 min, ΔG i = 104.47 kJ mol -1。同时,固定化酶的最适温度为60℃,km = 11.57 mg mL-1底物,vmax =3.37 μmolemL-1min-1, ki = 0.0045 min-1, t1 /2 = 154.00 min, ΔG i = 108.17 kJ mol -1。6次循环后,固定化酶的剩余活性为38%。α-淀粉酶固定化后的半衰期增加,其稳定性的提高是游离酶的4倍。
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引用次数: 9
Diets Rich in Olive Oil, Palm Oil, or Lard Alter Mitochondrial Biogenesis and Mitochondrial Membrane Composition in Rat Liver 富含橄榄油、棕榈油或猪油的饮食改变大鼠肝脏线粒体生物发生和线粒体膜组成
IF 3 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2022-02-21 DOI: 10.1155/2022/9394356
Y. F. Djohan, M. Camara-Cissé, G. Fouret, B. Bonafos, B. Jover, J. Cristol, C. Coudray, C. Feillet-Coudray, É. Badia
Palm oil (crude or refined) and lard are rich in SFA, while olive oil is rich in polyunsaturated fatty acids. SFA are considered harmful to health, while polyunsaturated fatty acids are beneficial to health. The aim of this study was to determine the effect of diets rich in crude PO, refined PO, OO, or lard on the mitochondrial membrane, the activity of mitochondrial respiratory chain complexes, and mitochondrial biogenesis. This was an experimental study in male Wistar rats fed a diet containing 30% of each oil. Rats had free access to food and water. After being fed for 12 weeks, animals were sacrificed and liver mitochondria were collected. This collection was used to determine membrane potential and ROS production, membrane phospholipid and fatty acid composition, citrate synthase activity and respiratory chain complex, cardiolipin synthase protein expression, and expression of selected genes involved in mitochondrial biogenesis. We found that diets rich in olive oil, palm oil, or lard altered mitochondrial biogenesis by significantly decreasing Pgc1α gene expression and altered the fatty acid composition of rat liver mitochondrial membrane PL.
棕榈油(原油或精炼)和猪油富含SFA,而橄榄油富含多不饱和脂肪酸。SFA被认为对健康有害,而多不饱和脂肪酸则对健康有益。本研究的目的是确定饲粮中富含粗PO、精制PO、OO或猪油对线粒体膜、线粒体呼吸链复合物活性和线粒体生物发生的影响。这是一项实验研究,雄性Wistar大鼠喂食含有每种油30%的食物。老鼠可以自由获取食物和水。饲养12周后处死动物,收集肝脏线粒体。这些收集物用于测定膜电位和ROS生成、膜磷脂和脂肪酸组成、柠檬酸合成酶活性和呼吸链复合体、心磷脂合成酶蛋白表达以及参与线粒体生物发生的选定基因的表达。我们发现,富含橄榄油、棕榈油或猪油的饮食通过显著降低Pgc1α基因表达和改变大鼠肝脏线粒体膜PL的脂肪酸组成来改变线粒体生物发生。
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引用次数: 2
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Biochemistry Research International
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