共查询到20条相似文献,搜索用时 15 毫秒
1.
Juliane Zimmerling Dirk Tischler Carolin Großmann Michael Schlömann Michel Oelschlägel 《Applied biochemistry and biotechnology》2017,182(3):1095-1107
Herein, different dehydrogenases (DH) were characterized by applying a novel two-step enzyme assay. We focused on the NAD(P)+-dependent phenylacetaldehyde dehydrogenases because they produce industrially relevant phenylacetic acids, but they are not well studied due to limited substrate availability. The first assay step comprises a styrene oxide isomerase (440 U mg?1 protein) which allows the production of pure phenylacetaldehydes (>70 mmol L?1) from commercially available styrene oxides. Thereafter, a DH of interest can be added to convert phenylacetaldehydes in a broad concentration range (0.05 to 1.25 mmol L?1). DH activity can be determined spectrophotometrically by following cofactor reduction or alternatively by RP-HPLC. This assay allowed the comparison of four aldehyde dehydrogenases and even of an alcohol dehydrogenase with respect to the production of phenylacetic acids (up to 8.4 U mg?1 protein). FeaB derived from Escherichia coli K-12 was characterized in more detail, and for the first time, substituted phenylacetaldehydes had been converted. With this enzyme assay, characterization of dehydrogenases is possible although the substrates are not commercially available in sufficient quality but enzymatically producible. The advantages of this assay in comparison to the former one are discussed. 相似文献
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Dr. Giuseppe Pietricola Lesly Chamorro Dr. Micaela Castellino Diego Maureira Prof. Tonia Tommasi Prof. Simelys Hernndez Prof. Lorena Wilson Prof. Debora Fino Prof. Carminna Ottone 《ChemistryOpen》2022,11(11)
This study presents the immobilization with aldehyde groups (glyoxyl carbon felt) of alcohol dehydrogenase (ADH) and formate dehydrogenase (FDH) on carbon‐felt‐based electrodes. The compatibility of the immobilization method with the electrochemical application was studied with the ADH bioelectrode. The electrochemical regeneration process of nicotinamide adenine dinucleotide in its oxidized form (NAD+), on a carbon felt surface, has been deeply studied with tests performed at different electrical potentials. By applying a potential of 0.4 V versus Ag/AgCl electrode, a good compromise between NAD+ regeneration and energy consumption was observed. The effectiveness of the regeneration of NAD+ was confirmed by electrochemical oxidation of ethanol catalyzed by ADH in the presence of NADH, which is the no active form of the cofactor for this reaction. Good reusability was observed by using ADH immobilized on glyoxyl functionalized carbon felt with a residual activity higher than 60 % after 3 batches. 相似文献
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Shukor MY Rahman MF Shamaan NA Lee CH Karim MI Syed MA 《Applied biochemistry and biotechnology》2008,144(3):293-300
Molybdenum-reducing activity in the heterotrophic bacteria is a phenomenon that has been reported for more than 100 years.
In the presence of molybdenum in the growth media, bacterial colonies turn to blue. The enzyme(s) responsible for the reduction
of molybdenum to molybdenum blue in these bacteria has never been purified. In our quest to purify the molybdenum-reducing
enzyme, we have devised a better substrate for the enzyme activity using laboratory-prepared phosphomolybdate instead of the
commercial 12-phosphomolybdate we developed previously. Using laboratory-prepared phosphomolybdate, the highest activity is
given by 10:4-phosphomolybdate. The apparent Michaelis constant, K
m for the laboratory-prepared 10:4-phosphomolybdate is 2.56 ± 0.25 mM (arbitrary concentration), whereas the apparent V
max is 99.4 ± 2.85 nmol Mo-blue min−1 mg−1 protein. The apparent Michaelis constant or K
m for NADH as the electron donor is 1.38 ± 0.09 mM, whereas the apparent V
max is 102.6 ± 1.73 nmol Mo-blue min−1 mg−1 protein. The apparent K
m and V
max for another electron donor, NADPH, is 1.43 ± 0.10 mM and 57.16 ± 1.01 nmol Mo-blue min−1 mg−1 protein, respectively, using the same batch of molybdenum-reducing enzyme. The apparent V
max obtained for NADH and 10:4-phosphomolybdate is approximately 13 times better than 12-phoshomolybdate using the same batch
of enzyme, and hence, the laboratory-prepared phosphomolybdate is a much better substrate than 12-phoshomolybdate. In addition,
10:4-phosphomolybdate can be routinely prepared from phosphate and molybdate, two common chemicals in the laboratory. 相似文献
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《Chemistry & biology》2014,21(11):1423-1432
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A natural proline‐rich tetrapeptide cyclo‐prolyl‐leucyl‐prolyl‐phenylalanyl was prepared employing solution‐phase method of peptide synthesis through coupling of dipeptide fragments Boc‐l‐Pro‐l‐Leu‐OH and l‐Pro‐l‐Phe‐OMe which utilizes diisopropylcarbodiimide (DIPC) as coupling agent and N‐methylmorpholine (NMM) as the base. Deprotection of linear tetrapeptide unit followed by its cyclization provided a cyclopeptide, identical in all aspects to the natural molecule. Pharmacological evaluation showed cytotoxic, antifungal and antihelmintic potential of synthesized peptide against Dalton's Lymphoma Ascites (DLA) and Ehrlich's Ascites Carcinoma (EAC) cell lines, pathogenic dermatophytes and earthworms. 相似文献
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Meng Gao Zhenzhen Wang Huizhen Zheng Li Wang Shujuan Xu Xi Liu Wei Li Yanxia Pan Weili Wang Xiaoming Cai Ren'an Wu Dr. Xingfa Gao Dr. Ruibin Li 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2020,132(9):3647-3652
While dehydrogenases play crucial roles in tricarboxylic acid (TCA) cycle of cell metabolism, which are extensively explored for biomedical and chemical engineering uses, it is a big challenge to overcome the shortcomings (low stability and high costs) of recombinant dehydrogenases. Herein, it is shown that two-dimensional (2D) SnSe is capable of mimicking native dehydrogenases to efficiently catalyze hydrogen transfer from 1-(R)-2-(R′)-ethanol groups. In contrary to susceptible native dehydrogenases, lactic dehydrogenase (LDH) for instance, SnSe is extremely tolerant to reaction condition changes (pH, temperature, and organic solvents) and displays extraordinary reusable capability. Structure–activity analysis indicates that the single-atom structure, Sn vacancy, and hydrogen binding affinity of SnSe may be responsible for their catalytic activity. Overall, this is the first report of a 2D SnSe nanozyme to mimic key dehydrogenases in cell metabolism. 相似文献
7.
Condenced aromatics in the environment are harmful to the organisms for its carcinogenesis. When it is metabolized in organism, several kinds of radicals and active oxygen intermediates such as superoxide radical anion (O2-) and hydrogen peroxide would produce. These active oxygen intermediates may cause DNA damage. Catalase (CAT), a major endogenous antioxidant, presents in all aerobes and has a crucial function in protecting living organisms against the unrestrained reactivity of H2O2. When the amount of H2O2 generated in the body increase due to the outside factors, the activity of the CAT will be induced to rise significantly. Thus the activity of CAT, as a biomarker, may reflect die exist of condensed aromatics in the environment. Now there are many methods to detect CAT, such as spectrophotometric methods, titration,electrochemical process, volumetric analysis, but they are not sensitive enough for direct detection of CAT in marine biological samples. 相似文献
8.
海洋沉积物中不同结合态无机碳的测定 总被引:12,自引:0,他引:12
作为全球碳的重要源与汇的海洋沉积物,在碳循环中起着重要的作用。探讨沉积物中无机碳的行为对深入了解海洋碳循环具有重要意义。本研究根据沉积物中无机碳在不同溶剂中的溶解能力不同,利用顺序浸提法,将沉积物中的无机碳分为:NaCl相(Ⅰ)、氨水相(Ⅱ)、氢氧化钠相(Ⅲ)、盐酸羟胺相(Ⅳ)和盐酸相(Ⅴ)。通过大量实验确定了不同相的最佳浸取条件。在Ⅰ~Ⅲ相,分别将浸取剂和样品加入塑料离心管中,将离心管密封后放在震荡器上震荡2h,然后离心分离,并用水洗涤残渣,将洗涤液并入浸取液,最后用容量法测定其中的无机碳;在Ⅳ,Ⅴ相,将第Ⅲ步残渣转移到锥形瓶中,加入浸取剂后用高纯N2将所产生的CO2吹出,并用饱和Ba(OH)2溶液吸收,最后用容量法测定无机碳。该方法具有较好的精密度。 相似文献
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Noora Barzkar Ruilong Sheng Muhammad Sohail Saeid Tamadoni Jahromi Olga Babich Stanislav Sukhikh Reza Nahavandi 《Molecules (Basel, Switzerland)》2022,27(11)
The cell wall of brown algae contains alginate as a major constituent. This anionic polymer is a composite of β-d-mannuronate (M) and α-l-guluronate (G). Alginate can be degraded into oligosaccharides; both the polymer and its products exhibit antioxidative, antimicrobial, and immunomodulatory activities and, hence, find many commercial applications. Alginate is attacked by various enzymes, collectively termed alginate lyases, that degrade glycosidic bonds through β-elimination. Considering the abundance of brown algae in marine ecosystems, alginate is an important source of nutrients for marine organisms, and therefore, alginate lyases play a significant role in marine carbon recycling. Various marine microorganisms, particularly those that thrive in association with brown algae, have been reported as producers of alginate lyases. Conceivably, the marine-derived alginate lyases demonstrate salt tolerance, and many are activated in the presence of salts and, therefore, find applications in the food industry. Therefore, this review summarizes the structural and biochemical features of marine bacterial alginate lyases along with their applications. This comprehensive information can aid in the expansion of future prospects of alginate lyases. 相似文献
12.
Meng Gao Zhenzhen Wang Huizhen Zheng Li Wang Shujuan Xu Xi Liu Wei Li Yanxia Pan Weili Wang Xiaoming Cai Ren'an Wu Xingfa Gao Ruibin Li 《Angewandte Chemie (International ed. in English)》2020,59(9):3618-3623
While dehydrogenases play crucial roles in tricarboxylic acid (TCA) cycle of cell metabolism, which are extensively explored for biomedical and chemical engineering uses, it is a big challenge to overcome the shortcomings (low stability and high costs) of recombinant dehydrogenases. Herein, it is shown that two‐dimensional (2D) SnSe is capable of mimicking native dehydrogenases to efficiently catalyze hydrogen transfer from 1‐(R)‐2‐(R′)‐ethanol groups. In contrary to susceptible native dehydrogenases, lactic dehydrogenase (LDH) for instance, SnSe is extremely tolerant to reaction condition changes (pH, temperature, and organic solvents) and displays extraordinary reusable capability. Structure–activity analysis indicates that the single‐atom structure, Sn vacancy, and hydrogen binding affinity of SnSe may be responsible for their catalytic activity. Overall, this is the first report of a 2D SnSe nanozyme to mimic key dehydrogenases in cell metabolism. 相似文献
13.
Khandaker Md Sharif Uddin Imam Yu Tian Fengjiao Xin Yingying Xie Boting Wen 《Molecules (Basel, Switzerland)》2022,27(21)
Lung cancer, especially adenocarcinoma, is the second most occurring and highest fatality-causing cancer worldwide. Many natural anticancer compounds, such as sesquiterpene lactones (SLs), show promising anticancer properties. Herein, we examined Lactucin, an SL from the plant Cichorium intybus, for its cytotoxicity, apoptotic-inducing, cell cycle inhibiting capacity, and associated protein expression. We also constructed a biotinylated Lactucin probe to isolate interacting proteins and identified them. We found that Lactucin stops the proliferation of A549 and H2347 lung adenocarcinoma cell lines while not affecting normal lung cell MRC5. It also significantly inhibits the cell cycle at G0/G1 stage and induces apoptosis. The western blot analysis shows that Lactucin downregulates the MAPK pathway, cyclin, and cyclin-dependent kinases, inhibiting DNA repair while upregulating p53, p21, Bax, PTEN, and downregulation of Bcl-2. An increased p53 in response to DNA damage upregulates p21, Bax, and PTEN. In an activity-based protein profiling (ABPP) analysis of A549 cell’s protein lysate using a biotinylated Lactucin probe, we found that Lactucin binds PGM, PKM, and LDHA PDH, four critical enzymes in central carbon metabolism in cancer cells, limiting cancer cells in its growth; thus, Lactucin inhibits cancer cell proliferation by downregulating the MAPK and the Central Carbon Metabolism pathway. 相似文献
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Prof. Dr. Musa M. Musa 《ChemistryOpen》2022,11(4):e202100251
Biocatalytic production of both enantiomers of optically active alcohols with high enantiopurities is of great interest in industry. Alcohol dehydrogenases (ADHs) represent an important class of enzymes that could be used as catalysts to produce optically active alcohols from their corresponding prochiral ketones. This review covers examples of the synthesis of optically active alcohols using ADHs that exhibit anti-Prelog stereopreference. Both wild-type and engineered ADHs that exhibit anti-Prelog stereopreference are highlighted. 相似文献
15.
海洋用钢在海底泥土区常承受具大的应力 ,而应力将对海洋用钢的腐蚀破坏行为产生影响 .本文选用一种常用的海洋用钢 ,采集含有硫酸盐还原菌 (SRB)的天然海泥 ,研究试样在海泥中的腐蚀行为 ,以及试样在有裂纹和无裂纹存在时的电流响应特征 相似文献
16.
血红蛋白在壳聚糖修饰碳纳米管上的电化学特性及对过氧化氢的电催化分析 总被引:3,自引:2,他引:1
用壳聚糖对多壁碳纳米管进行修饰,构建了一种用于固定血红蛋白的新型复合材料,并研究了血红蛋白在该碳纳米管上的电化学性质及其对过氧化氢的电催化活性.扫描电镜结果表明,壳聚糖修饰的多壁碳纳米管呈单一的纳米管状,并能均匀分散在玻碳电极表面.紫外光谱分析表明血红蛋白在该复合膜内能很好地保持其原有的二级结构.将该材料固定在玻碳电极上后,血红蛋白能成功地实现其直接电化学.根据峰电位差随着扫描的变化,计算得到血红蛋白在壳聚糖修饰的碳纳米管膜上的电荷转移系数为0.57,表观电子转移速率常数为7.02 s-1.同时,该电极对过氧化氢显示出良好的催化性能,电流响应信号与H2O2浓度在1.0×10-6 ~1.5×10-3 mol/L间呈线性关系,检出限为5.0×10-7 mol/L.修饰电极显示了良好的稳定性. 相似文献
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Temperature increase due to resistive electrical heating is an inherent limitation of capillary electrophoresis (CE). Active cooling systems are used to decrease the temperature of the capillary, but their capacity is limited; and in addition, they leave "hot spots" at the detection interface and at the capillary ends. Until recently, the matter was complicated by the lack of a fast and generic method for temperature determination in efficiently and inefficiently cooled regions of the capillary. Our group recently introduced such a method, termed "Universal Method for determining Electrolyte Temperatures" (UMET). UMET is a probe-less approach that requires only measuring current versus voltage for different voltages and processing the data using an iterative algorithm. Here, we apply UMET to develop a Simplified Universal Method of Temperature Determination (SUMET) for a CE instrument with a forced-air cooling system using an Agilent 7100 CE instrument (Agilent Technologies, Saint Laurent, Quebec, Canada) as an example. We collected a wide set of empirical voltage-current data for a variety of buffers and capillary diameters. We further constructed empirical equations for temperature calculation in efficiently and inefficiently cooled parts of the capillary that require only the data from a single 1-min voltage-current measurement. The equations are specific for the Agilent 7100 CE instrument (Agilent Technologies) but can be applied to all kinds of capillaries and buffers. Similar SUMET approaches can be developed for other CE instruments with forced-air cooling using our approach. 相似文献
20.
Dr. Pengfei Xia Prof. Shaowen Cao Dr. Bicheng Zhu Mingjin Liu Dr. Miusi Shi Prof. Jiaguo Yu Prof. Yufeng Zhang 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2020,132(13):5256-5263
Constructing heterojunctions between two semiconductors with matched band structure is an effective strategy to acquire high-efficiency photocatalysts. The S-scheme heterojunction system has shown great potential in facilitating separation and transfer of photogenerated carriers, as well as acquiring strong photoredox ability. Herein, a 0D/2D S-Scheme heterojunction material involving CeO2 quantum dots and polymeric carbon nitride (CeO2/PCN) is designed and constructed by in situ wet chemistry with subsequent heat treatment. This S-scheme heterojunction material shows high-efficiency photocatalytic sterilization rate (88.1 %) towards Staphylococcus aureus (S. aureus) under visible-light irradiation (λ≥420 nm), which is 2.7 and 8.2 times that of pure CeO2 (32.2 %) and PCN (10.7 %), respectively. Strong evidence of S-scheme charge transfer path is verified by theoretical calculations, in situ irradiated X-ray photoelectron spectroscopy, and electron paramagnetic resonance. 相似文献