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411.
Carboxylesterase and sorbitol dehydrogenase are separated by non-denaturing two-dimensional electrophoresis (2-DE) of isoelectric focusing separation using 5% carrier ampholyte (pH 6-8) and 1.25% carrier ampholyte (pH 3-10) and size separation. Furthermore, activities of sorbitol, malate and lactate dehydrogenases are sequentially examined when the enzymes are separated by 2-DE and are sequentially reacted to sorbitol, malic and lactic acid, respectively, in the presence of nicotinamide adenine dinucleotide, nitro blue tetrazolium and phenazine methosulphate. Several kinds of enzymes including lactate dehydrogenize isozymes can be simultaneously separated using 2-DE. Furthermore, the binding differences between lactate dehydrogenase isozymes and concanavalin A (con A) can be examined using a combination of 2-DE and non-denaturing stacking gel electrophoresis. The results of this study indicate that non-denaturing 2-DE can be applied to both enzyme separation and isozyme heterogeneity analysis. 相似文献
412.
Computational analyses of mammalian lactate dehydrogenases: Human, mouse, opossum and platypus LDHs 总被引:1,自引:0,他引:1
Computational methods were used to predict the amino acid sequences and gene locations for mammalian lactate dehydrogenase (LDH) genes and proteins using genome sequence databanks. Human LDHA, LDHC and LDH6A genes were located in tandem on chromosome 11, while LDH6B and LDH6C genes were on chromosomes 15 and 12, respectively. Opossum LDHC and LDH6B genes were located in tandem with the opossum LDHA gene on chromosome 5 and contained 7 (LDHA and LDHC) or 8 (LDH6B) exons. An amino acid sequence prediction for the opossum LDH6B subunit gave an extended N-terminal sequence, similar to the human and mouse LDH6B sequences, which may support the export of this enzyme into mitochondria. The platypus genome contained at least 3 LDH genes encoding LDHA, LDHB and LDH6B subunits. Phylogenetic studies and sequence analyses indicated that LDHA, LDHB and LDH6B genes are present in all mammalian genomes examined, including a monotreme species (platypus), whereas the LDHC gene may have arisen more recently in marsupial mammals. 相似文献
413.
Lothar Ketterer 《Analytica chimica acta》2010,673(1):54-59
An amperometric sensor for the detection of cyanide integrated in a flow injection analysis (FIA) system is presented. The specificity of the sensor for cyanide is achieved by a reaction cascade involving two enzymes, cyanidase (EC 3.5.5.1) and formate dehydrogenase (FDH, EC 1.2.1.2). The limit of detection (LOD) and the limit of quantification (LOQ) were determined to be in the low micromolar range. The sensor showed very good long term stability and the linear range of detection extends from 0.7 up to 800 M. The sensor’s ability to measure cyanide in complex matrices, such as plant extracts, is demonstrated. Although the sensor is susceptible to formate and to a lesser extent to thiocyanate, sulfide does not interfere significantly. 相似文献
414.
415.
《Analytical letters》2012,45(18):1589-1606
Abstract The spectrofluorimetric determination of kinetic parameters (Km, V and kobs) of conjugated bile acids with 3α-hydroxysteroid dehydrogenase from mutant Ps. Testosteroni is described. Under optimal pH conditions and pseudo- first order kinetics([β-NAD+)20 KmNAD+) the kinetic parameters of bile acids in different buffers are determined. The bile acids exhibit larger kinetic differences in glycine and Tris buffer compared to those measured in pvrophosphate buffer. From these data it is concluded that under certain conditions the differential kinetic determination of one bile acid in the presence of others is possible without prior separation. 相似文献
416.
Zhou W Capello M Fredolini C Racanicchi L Piemonti L Liotta LA Novelli F Petricoin EF 《Electrophoresis》2012,33(12):1850-1854
L-lactate dehydrogenase (LDH) converts pyruvate to lactate when oxygen is absent or in short supply, and the enzyme plays a crucial role in cancer metabolism. The functions of many mammalian proteins are modulated by posttranslational modifications (PTMs), and it has been reported that LDH was subjected to several PTMs, including phosphorylation, acetylation, and methylation. In this present work, we characterized the PTMs of LDH from pancreatic ductal adenocarcinoma (PDAC) cells by electrophoresis and mass spectrometry, and identified 13 O-methylated residues from the enzyme. In addition, our qualitative analysis revealed differential methylation of LDH from normal duct cells. The preliminary findings from this study provide important biochemical information toward further understanding of the LDH modifications and their functional significance in pathophysiological processes of pancreatic cancer. 相似文献
417.
《Analytical letters》2012,45(12):2205-2215
Abstract A simple enzymatic end point method for the determination of L-phenylalanine and 3-phenylpyruvate has been developed. The assay is based on the spectrophotometry determination of NADH formed or degraded in the reaction catalyzed with Thermoactinomyces intermedins phenylalanine dehydrogenase. This method enables the simple and selective determination of L-phenylalanine and 3-phenylpyruvate based on the high substrate specificity of the enzyme. The assay is sensitive in the range of 0.015?0.15 umol of L-phenylalanine and 3-phenylpyruvate. 相似文献
418.
Riccardo Balzan Laetitia Fernandes Laetitia Pidial Arnaud Comment Bertrand Tavitian Paul R. Vasos 《Magnetic resonance in chemistry : MRC》2017,55(6):579-583
Pyruvate membrane crossing and its lactate dehydrogenase‐mediated conversion to lactate in cells featuring different levels of expression of membrane monocarboxylate transporters (MCT4) were probed by dissolution dynamic nuclear polarization‐enhanced NMR. Hyperpolarized 13C‐1‐labeled pyruvate was transferred to suspensions of rodent tumor cell carcinoma, cell line 39. The pyruvate‐to‐lactate conversion rate monitored by dissolution dynamic nuclear polarization‐NMR in carcinoma cells featuring native MCT4 expression level was lower than the rate observed for cells in which the human MCT4 gene was overexpressed. The enzymatic activity of lactate dehydrogenase was also assessed in buffer solutions, following the real‐time pyruvate‐to‐lactate conversion speeds at different enzyme concentrations. Copyright © 2016 John Wiley & Sons, Ltd. 相似文献
419.
《Electroanalysis》2017,29(11):2646-2655
Guanine‐ionic liquid derived ordered mesoporous carbon (GIOMC) decorated with gold nanoparticles was used as electrocatalyste for NADH oxidation and electrochemical platform for immobilization of glucose dehydrogenase (GDH) enzyme. The resulting GIOMC/AuNPs on the glassy carbon electrode can be used as novel redox‐mediator free for NADH sensing and this integrated system (GIOMC/AuNPs/GDH) shows excellent electrocatalytic activity toward glucose oxidation. Furthermore, the ionic liquid derived ordered mesoporous carbon derivate with Ph‐SO3H (IOMC‐PhSO3H) decorated with AuNPs has been developed to bilirubin oxidase enzyme (BOD) immobilization and the GC/IOMC‐PhSO3H/BOD integrated system shows excellent bioelectrocatalytic activity toward oxygen reduction reaction. The proposed mesostructured platforms decorated by AuNPs have been developed to enhance mass transfer and charge transfer from biocatalyst to electrode, leading these bioanode and biocathode used for biofuel cell assembly. Integration designed bioanode and biocathode yielding a membrane‐less glucose/O2 biofuel cell with power density of 33 (mW.cm−2) at 257 mV. The open circuit voltage of this biofuel cell and maximum produced current density were 508 mV and 0.252 (mA.cm−2) respectively. 相似文献
420.
用吐温80—盐—水液—固萃取体系从兔肌中分离纯化乳酸脱氢酶 总被引:5,自引:0,他引:5
用吐温80-盐-水液-固萃取体系从兔肌中分离纯化乳酸脱氢酶*柳畅先邓江天雷锦桂游文淑李步海(中南民族学院化学系武汉430074)关键词液-固萃取吐温80乳酸脱氢酶中图分类号O658.2非有机溶剂液-固萃取体系是一种新的萃取体系,其特点之一是聚合物固相... 相似文献