共查询到20条相似文献,搜索用时 15 毫秒
1.
Elena Fernández-Álvaro Robert Kourist Julia Winter Dominique Böttcher Klaus Liebeton Christian Naumer Jürgen Eck Christian Leggewie Karl-Erich Jaeger Wolfgang Streit Uwe T. Bornscheuer 《Microbial biotechnology》2010,3(1):59-64
Enantiomerically pure β-arylalkyl carboxylic acids are important synthetic intermediates for the preparation of a wide range of compounds with biological and pharmacological activities. A library of 83 enzymes isolated from the metagenome was searched for activity in the hydrolysis of ethyl esters of three racemic phenylalkyl carboxylic acids by a microtiter plate-based screening using a pH-indicator assay. Out of these, 20 enzymes were found to be active and were subjected to analytical scale biocatalysis in order to determine their enantioselectivity. The most enantioselective and also enantiocomplementary biocatalysts were then used for preparative scale reactions. Thus, both enantiomers of each of the three phenylalkyl carboxylic acids studied could be obtained in excellent optical purity and high yields. 相似文献
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Hui Jiang Rohini Sidhu Hideji Fujiwara Marc De Meulder Ronald de Vries Yong Gong Mark Kao Forbes D. Porter Nicole M. Yanjanin Nuria Carillo-Carasco Xin Xu Elizabeth Ottinger Myra Woolery Daniel S. Ory Xuntian Jiang 《Journal of lipid research》2014,55(7):1537-1548
2-Hydroxypropyl-β-cyclodextrin (HP-β-CD), a widely used excipient for drug formulation, has emerged as an investigational new drug for the treatment of Niemann-Pick type C1 (NPC1) disease, a neurodegenerative cholesterol storage disorder. Development of a sensitive quantitative LC-MS/MS assay to monitor the pharmacokinetics (PKs) of HP-β-CD required for clinical trials has been challenging owing to the dispersity of the HP-β-CD. To support a phase 1 clinical trial for ICV delivery of HP-β-CD in NPC1 patients, novel methods for quantification of HP-β-CD in human plasma and cerebrospinal fluid (CSF) using LC-MS/MS were developed and validated: a 2D-LC-in-source fragmentation-MS/MS (2D-LC-IF-MS/MS) assay and a reversed phase ultra performance LC-MS/MS (RP-UPLC-MS/MS) assay. In both assays, protein precipitation and “dilute and shoot” procedures were used to process plasma and CSF, respectively. The assays were fully validated and in close agreement, and allowed determination of PK parameters for HP-β-CD. The LC-MS/MS methods are ∼100-fold more sensitive than the current HPLC assay, and were successfully employed to analyze HP-β-CD in human plasma and CSF samples to support the phase 1 clinical trial of HP-β-CD in NPC1 patients. 相似文献
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Δ1-Tetrahydrocannabinol was found to inhibit the action of esterases derived from rat adrenal and luteinized ovary on exogenous cholesteryl palmitate. The drug was effective at a dose of 3.2μM causing greater than 30% inhibition; at 16μM almost complete inhibition occured. These findings are similar to those we have recently reported with mouse Leydig cells (1) showing that this is an effect common to steroidogenic tissues and raising the possibility that a variety of endocrine effects of this drug may be due to direct action on these tissues. 相似文献
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Biochemical properties,homology, and genetic variation of Drosophila “nonspecific” esterases 总被引:2,自引:0,他引:2
The biochemical properties and tissue distribution of two major, soluble "nonspecific" esterases have been studied in Drosophila melanogaster, D. pseudoobscura, and related species. The "alpha-like" activity is due to a monomer enzyme (MW congruent to 60 kd) having a nonspecific tissue distribution, which was inhibited by p-hydroxymercuribenzoate (5 X 10(-4)M) plus eserine (1 X 10(-5)M) and was relatively unstable during polyacrylamide gel electrophoresis. Electrophoretograms of this enzyme could be enhanced by treating gels with beta-mercaptoethanol before staining. This procedure allowed the identification of a new alpha-esterase (Est-4) in D. pseudoobscura. The "beta-like" esterase activity (EC 3.1.1.1) is due to a dimer (MW congruent to 120 kd) in most Drosophila species. D. melanogaster and its siblings (D. simulans and D. mauritiana) were exceptions in which this enzyme had an unusual tissue distribution (increased activity in the male reproductive system) and was a monomer (MW congruent to 60 kd). Differences in the genetic variability of these esterases are discussed and interpreted by a population expansion model rather than by differences in biochemical properties of enzyme forms. 相似文献
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Craig B. Faulds 《Phytochemistry Reviews》2010,9(1):121-132
The role of feruloyl esterases in plant wall development, in gut health, and in the breakdown of plant biomass for the production
of bioactive phytochemicals and biofuel is covered in this review. These enzymes have potential roles in stomatal cell function
and the phenolic substitutions and cross-linkages between plant cell wall components. As more plant genomes are sequenced,
the role of ferulic acid and feruloyl esterases in planta may be better understood. In human and ruminal digestion, these enzymes are important to de-esterify dietary fibre, releasing
hydroxycinnamates and derivatives which have been shown to have positive health effects, such as antioxidant, anti-inflammatory
and anti-microbial activities. They are also involved in colonic fermentation where their extracellular and intracellular
activities in the microbiota improve the breakdown of polysaccharides and increase microbial production of short chain fatty
acids. Their specificity can also be employed to synthesize bioactive compounds for cosmetic and health applications. The
enzymatic disassembly of cereal straws is greatly enhanced when feruloyl esterase activity is present, although the substrate
specificity of the esterase appears to have some bearing on its optimal application. The involvement of feruloyl esterases
in the improved enzymatic and microbial saccharification of cereal-derived material demonstrates a high importance for these
enzymes in animal feed preparation and bioalcohol production. 相似文献
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Michael J. Bailey 《Biotechnology letters》1981,3(12):695-700
The effect of -glucosidase on three assays for cellulolytic enzymes, i. e. the activities against dyed Avicel, hydroxyethylcellulose (HEC) and filter paper (FPU), was studied using cellulase enzyme derived from Trichoderma reesei VTT-D-80133. The dyed Avicel and HEC assays were only slightly affected by -glucosidase, whereas the FPU assay was linearly dependent on the level of -glucosidase over a wide range of activity of this enzyme. 相似文献
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Summary The subcellular localization of esterases and acid phosphatases in described for cells from roots of Vicia faba. The possible mode of transport of the esterases from a cytoplasmic site of synthesis to the cell wall is discussed. 相似文献
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I. Y. Rauschenbach L. B. Golosheikina L. S. Korochkina L. I. Korochkin 《Biochemical genetics》1977,15(5-6):531-548
From analysis of the properties of the pupal esterase (p-esterase) in Drosophila virilis, it is concluded that it is heat stable, its electrophoretic detection depends on culture density, its expression is stage specific, and it is not a variant of esterase 2. It was also demonstrated that p-esterase, like esterase 6, is activated by injections of the juvenile hormone into larvae. Heat treatment of heat-resistant D. virilis stocks led to decreased activities of the juvenile hormone dependent esterases but did not affect those of the heat-sensitive stocks. It is suggested that heat resistance in D. virilis is related to some functional features of the system of modifier genes controlling the phenotypic expression of esterases. 相似文献
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Because of increasing awareness and legislative demands, there is a demand for the development and use of biological assays
for the assessment of the toxicity of chemicals, environmental samples. Recently, a growing number of bacterial reporter assays
have been developed and implemented. Nevertheless, little data is published on the performance of these assays in terms of
analytical parameters. We present results on a battery of 14 transgenic Escherichia coli strains carrying different promoter::reporter fusions. Growth characteristics and basal expression levels were modeled and
fitted, data show that growth curves should be taken into account during test development. Our study shows that the induction
profiles reflect the mode of action, e.g., paraquat clearly induces the soxRS operon. The sensitivity of the assay compares well to that of whole organism tests, e.g., fish and Daphnia for polar organics.
Metal toxicity is detected less efficiently, e.g., cadmium is detected near the LC50 of carp, considered a relatively insensitive
species towards cadmium. The assay variability ranges from 10 to 40% depending on the strain, comparable to that of other
bioassays. The variability was shown to be determined by the intrinsic traits of the promoter–strain combination, not by operating
conditions.
Electronic supplementary material Supplementary material is available for this article at and is accessible for authorized users. 相似文献
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Green Pattie S. Perez Evelyn J. Calloway Tammy Simpkins James W. 《Brain Cell Biology》2000,29(5-6):419-423
17β-estradiol (βE2) has been shown to attenuate the toxicity of β-amyloid peptides (Aβ) in neuronal cultures with the effective concentration of βE2 ranging from low nM to high μM. This study compares the effective neuroprotective concentration of βE2 against both Aβ-mediated toxicity in a human neuroblastoma cell line, SK-N-SH using cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) as an endpoint to the effective βE2 concentration obtained using a calcein acetoxymethyl ester (calcein AM) viability assay. The minimum βE2 concentration required for protection varied 1000-fold between the two viability assays with 1 nM βE2 conferring significant protection in the calcein AM assay but 1 μM βE2 required for significant protection in the MTT assay. Interestingly, the maximal inhibition of MTT reduction occured at sub-toxic Aβ concentrations and did not correlate with other markers of cellular viability including calcein fluorescence, dye exclusion (propidium iodide or trypan blue), cellular ATP levels, or reduction of another tetrazolium dye, 5-(3-carboxymethoxyphenyl)-2-(4,5-dimethylthiazolyl)-3-(4-sulfophenyl) tetrazolium (MTS). By contrast, there was no difference between the MTT and calcein AM assays with respect to H2O2 toxicity or the neuroprotective effectiveness of 10 nM βE2 against H2 2 toxicity. These results indicate that low concentrations of βE2 can attenuate Aβ and H2O2 toxicity in a human neuroblastoma cell line. Further, these results suggest that the MTT assay is not an appropriate assay for the determination of βE2-mediated attenuation of Aβ toxicity. 相似文献
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The finding that a variety of chemicals display oestrogenic activity has resulted in the development of in vitro and in vivo assays to assess oestrogenic activity. One such assay, the yeast oestrogen assay (YES) makes use of recombinant yeast cells that harbour an oestrogen receptor expression cassette and a reporter construct, coding for bgalactosidase. The induction mechanism starts with the binding of oestrogenic compounds to the oestrogen receptor. This complex activates the production of β-galactosidase. The β-galactosidase activity is thus a measure of the oestrogenic activity of chemical compounds. In the YES assay, the β-galactosidase activity may be quantified with the chromogenic substrate chlorophenol red-β-d-galactopyranoside (CPRG). In the present study it is reported that CPRG or its β-galactosidase degradation product chlorophenol red act in the YES as an oestrogenic compound itself. The implications of this finding are described. It is especially argued that chlorophenol red production after prolonged incubation of the assay might be misinterpreted as an oestrogenic effect of the test compound. 相似文献
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Aggregation of α-synuclein plays a crucial role in the pathogenesis of synucleinopathies, a group of neurodegenerative diseases including Parkinson disease (PD), dementia with Lewy bodies (DLB), diffuse Lewy body disease (DLBD) and multiple system atrophy (MSA). The common feature of these diseases is a pathological deposition of protein aggregates, known as Lewy bodies (LBs) in the central nervous system. The major component of these aggregates is α-synuclein, a natively unfolded protein, which may undergo dramatic structural changes resulting in the formation of β-sheet rich assemblies. In vitro studies have shown that recombinant α-synuclein protein may polymerize into amyloidogenic fibrils resembling those found in LBs. These aggregates may be uptaken and propagated between cells in a prion-like manner. Here we present the mechanisms and kinetics of α-synuclein aggregation in vitro, as well as crucial factors affecting this process. We also describe how PD-linked α-synuclein mutations and some exogenous factors modulate in vitro aggregation. Furthermore, we present a current knowledge on the mechanisms by which extracellular aggregates may be internalized and propagated between cells, as well as the mechanisms of their toxicity. 相似文献
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《生物化学与分子生物教育》2001,29(2):76-78
An interesting example of interference in the arginase assay is presented. This can be exploited to convey the importance of taking precautions and appropriate controls to the students of enzymology. 相似文献
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Kuz'min EV 《Genetika》2002,38(4):507-514
Electrophoresis in 7% polyacrylamide gel was used to analyze nonspecific esterases from different organs and tissues of Russian sturgeon. The basic enzymatic activity was observed the four zymogram zones. In sixteen cases the patterns of isozyme distribution in these zones allowed to consider them as allele systems (14 polymorphic and 2 monomorphic) corresponding to the four structural genes. The observed genotype frequencies of the nine of these systems (EST-1* of brain and intestine; EST-2* of serum and intestine; EST-3* of spleen and kidney; and EST-4* of spleen, brain, and heart) were largely concordant with Hardy-Weinberg proportions for codominant disomic inheritance. Genetic control of other polymorphic systems is unknown, but, high enzymatic activity of these esterases, sufficient for qualitative electrophretic detection, permits utilization of these polymorphisms for phenotypic monitoring of Russian sturgeon populations. 相似文献
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