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1.
Gender difference in human bisphenol A (BPA) concentrations was revealed by determining serum BPA. We studied the serum concentrations and the metabolism of BPA in rats by an HPLC system. Rat serum BPA concentrations were significantly higher in males (24.9+/-7.38 ng/ml, P=0.026, n=10) than in females (8.27+/-3.11 ng/ml, n=10), as in humans. The resultant enzyme reaction products of BPA glucuronidation in the rat liver microsomes fraction were analyzed by an HPLC system. The ratio of BPA glucuronidation in the microsome reaction was significantly higher (P=0.015) in female than in male rats. The mRNA expression of UDP-glucuronosyltransferase 2B1 (UGT2B1), an isoform of UGT related to BPA glucuronidation, in the rat liver was analyzed by a real-time quantitative RT-PCR. The relative expression level of UGT2B1 mRNA was significantly higher (P<0.001) in female than in male rat livers. The gender difference in serum BPA concentrations may be explained by the difference in clearance based on the UGT activities. 相似文献
2.
深山含笑是一种常绿乔木,具有较好的园林观赏价值,其叶片在每年春天新旧叶交替时,会陆续凋落而不能实现其经济价值;我们前期研究发现深山含笑成熟叶片具有较高的过氧化物酶活性,而过氧化物酶在内分泌干扰物生物酶法降解途径中发挥着重要作用。为充分开发深山含笑叶的经济价值,提高资源的利用效率,我们对深山含笑叶过氧化物酶进行了纯化并初步研究了其性质;在此基础上,进一步研究了深山含笑叶过氧化物酶在降解环境污染物双酚A上的特性。结果显示,经过非离子表面活性剂(吐温-80)抽提、双水相萃取、DEAE-Sepharose离子交换层析,首次从深山含笑叶片中快速分离提纯结合态过氧化物酶,相对可溶性过氧化物酶纯化倍数为55. 2倍,回收率为19%。该酶的比活力为18 999 U/mg蛋白,以愈创木酚为反应底物的最佳pH值为4. 5,最适温度为50℃;具有较宽的pH稳定范围,热稳定性较好,60℃以下不易失活。深山含笑叶过氧化物酶对BPA具有良好的清除能力,在pH4~7,温度30~40℃,H2O2/BPA摩尔浓度之比达0. 8条件下,经过3 h,10 000 U/L POD对0. 2 mmol/L BPA清除率达90%以上,深山含笑过氧化物酶在酚类物质生物酶法催化降解途径中具有潜在的重大利用价值。 相似文献
3.
The catalytic performance of chloroperoxidase (CPO) in peroxidation of 2, 2′-azinobis-(-3 ethylbenzothiazoline-6-sulfononic
acid) diammonium salt (ABTS) and oxidation of indole in a reverse micelle composed of surfactant-water-isooctane-pentanol
was investigated and optimized in this work. Some positive results were obtained as follows: the peroxidation activity of
CPO was enhanced 248% and 263%, while oxidation activity was enhanced 215% and 222% in cetyltrimethylammonium bromide (CTABr)
reverse micelle medium and dodecyltrimethylammonium bromide (DTABr) medium, respectively. Thermostability was also greatly
improved in reverse micelle: at 40°C, CPO essentially lost all its activity after 5 h incubation, while 58–76% catalytic activity
was retained for both reactions in the two reverse micelle media. At 50°C, about 44–75% catalytic activity remained for both
reactions in reverse micelle after 2 h compared with no observed activity in pure buffer under the same conditions. The enhancement
of CPO activity was dependent mainly on the surfactant concentration and structure, organic solvent ratio ( V
pentanol/ V
isooctane), and water content in the reverse micelle. The obtained kinetic parameters showed that the catalytic turnover frequency
( k
cat) was increased in reverse micelle. Moreover, the lower K
m and higher k
cat/ K
m demonstrated that both the affinity and specificity of CPO to substrates were improved in reverse micelle media. Fluorescence,
circular dichroism (CD) and UV–vis spectra assays indicated that a catalytically favorable conformation of enzyme was achieved
in reverse micelle, including the strengthening of the protein α-helix structure, and greater exposure of the heme prosthetic
group for easy access of the substrate in bulk solution. These results are promising in view of the industrial applications
of this versatile biological catalyst. 相似文献
5.
HAuCl 4 was reduced by sodium citrate to prepare 10 nm gold nanoparticles (AuNPs) that were modified by the bisphenol A aptamer (Apt) to obtain an aptamer–nanogold probe (Apt‐AuNP) for bisphenol A (BPA). The probes were aggregated nonspecifically to form large clusters, which showed a strong resonance light scattering (RLS) peak at 520 nm, under preparation conditions (pH 7.6 Na 2HPO 4‐NaH 2PO 4 buffer and ultrasonication). Upon addition of BPA, the probe reacted specifically to form dispersed BPA‐Apt‐AuNP conjugates that exhibited strong catalysis of the two particle reactions of glucose‐Cu(II) and hydrazine hydrochloride‐Cu(II) with a strong RLS peak at 360 nm and 510 nm respectively. When the BPA concentration increased, the RLS intensity at 360 nm and 510 nm increased respectively. Accordingly, two new and highly‐sensitive RLS methods were established for the detection of BPA, using the Apt‐AuNP catalytic amplification. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
6.
For an accurate determination of bisphenol A (BPA) in red blood cells (RBC), the effect of pH on the concentration of BPA was investigated. Also, BPA recovery using ferric heme, methemoglobin (metHb) and hematin, were investigated to confirm whether BPA binds to ferric heme. BPA recovery in hemolysate was high at alkaline pH and was very low at acidic pH where oxyHb changed to metHb. BPA recovery decreased dose-dependently in metHb and hematin, but inorganic iron ions did not influence the recovery. These results suggested that BPA could be bound to ferric heme in RBC. The use of glycine-NaOH buffer (pH 11) as well as plasma had the highest recovery (97%). BPA was not detected in red blood cells of healthy adult volunteers (n=6). In sheep blood contaminated with BPA, BPA was detected in both plasma and RBC (10 times lower than in plasma), indicating that BPA could have migrated from plasma into RBC. 相似文献
7.
The activity of lignin peroxidase (LiP) and the partition of its optimum substrate veratryl alcohol (VA) in sodium bis(2-ethylhexyl)sulfosuccinate (AOT)/isooctane/toluene/water reverse micelles were studied in this paper to understand the microheterogeneous effect of the medium on the catalytic properties of LiP hosted in the reverse micelle. Results showed that LiP from Phanerochaete chrysosporium could express its activity in the reverse micelles, but its activity depended, to a great extent, on the composition of the reverse micelles. Optimum activity occurred at a molar ratio of water to AOT (ω 0) of 11, a pH value of 3.6, and a volume ratio of isooctane to toluene of 7–9. Under optimum conditions, the half-life of LiP was circa 12 h. The dependence of LiP activity on the volume fraction of water in the medium (θ), at a constant ω 0 value of 11, indicated that VA was mainly solubilized in the pseudophase of the reverse micelle. Based on the pseudobiphasic model and the corresponding kinetic method, a linear line can be obtained in a plot of apparent Michaelis constant of VA vs θ, and the partition coefficient of VA between the pseudophase and the organic solvent phase was determined to be 35.8, which was higher than that (22.3) between bulk water and the corresponding mixed organic solvent. H 2O 2 inhibited LiP at concentrations higher than 80 μM; this concentration value seems to be different from that in aqueous solution (about 3 mM). The differences mentioned above should be ascribed to the microheterogeneity and the interface of the AOT reverse micelle. 相似文献
8.
Possible biotechnological applications of extreme halophilic enzymes are strongly determined by their high salt requirement of around 4 M NaCl. Consequently, the use of these in organic media seemed to be unlikely. However, we have succeeded in dissolving a halophilic enzyme, p-nitrophenylphosphate phosphatase from the archaeon Halobacterium salinarum, in an organic medium by creating a reverse micellar system with very low salt concentration. The enzyme retained its catalytic properties in reversed micelles made with an anionic surfactant (dioctyl sodium sulphosuccinate) or with a cationic surfactant (hexadecyltrimethylammonium bromide) in cyclohexane plus 1-butanol as co-surfactant. The dependence of the rate of hydrolysis of p-nitrophenylphosphate phosphate on the molar water/surfactant ratio (w(0) value) showed a bell-shaped curve for each surfactant system. Kinetic parameters were determined in each system. The enzymatic reaction appeared to follow Michaelis-Menten kinetics with the anionic surfactant only. The kinetic behaviour was determined at different concentrations of Mn(2+) in reversed micelles of dioctyl sodium sulphosuccinate as surfactant. 相似文献
9.
The oxidative conversion of aqueous BPA catalyzed by laccase from Trametes versicolor was conducted in a closed, temperature-controlled system containing buffer for pH control. The effects of medium pH, buffer concentration, temperature and mediators and the impacts of dissolved wastewater constituents on BPA conversion were investigated. The optimal pH for BPA conversion was approximately 5, with greater than half maximal conversion and good enzyme stability in the range of 4-7. The stability of the enzyme was not impacted by buffer concentration, nor was BPA conversion. Despite the observation that the enzyme tended to be inactivated at elevated temperatures, enhanced conversion of BPA was observed up until a reaction temperature of 45 degrees C. Of the mediators studied, ABTS was most successful at enhancing the conversion of BPA. Dissolved wastewater constituents that were studied included various inorganic salts, organic compounds and heavy metal ions. BPA conversion was inhibited in the presence of anions such as sulfite, thiosulfate, sulfide, nitrite and cyanide. The metal ions Fe(III) and Cu(II) and the halogens chloride and fluoride substantially suppressed BPA conversion, but the presence of selected organic compounds did not significantly reduce the conversion of BPA. 相似文献
10.
To explore the effects of maternal nutrition on offspring muscle characteristics, a total of 56 sows were assigned to one of the four dietary groups during gestation: control (CON), or control diets supplemented with methyl donor (MET), bisphenol A (BPA), and combined BPA and MET (BPA+MET). Compared with CON offspring, MET offspring showed a higher meat redness value, but lower glycogen content in the longissimus thoracis (LT). Moreover, compared with CON offspring, MET offspring showed lower LT glycogen synthase ( GS) mRNA levels at birth and the finishing stage, and increased methylation at the GS promoter. Prenatal BPA exposure reduced the pH and redness value of meat, but increased the lightness value, lactate content, glycolytic potential and lactate dehydrogenase (LDH) enzyme activity in the LT muscle. Prenatal BPA exposure increased LDH mRNA levels in the LT muscle at birth and the finishing stage, and reduced methylation at the LDH promoter. Thus, maternal MET affects muscle GS and LDH expression via DNA methylation, thereby resulting in persistent effects on pork quality. 相似文献
11.
The enzymatic production of tryptophan from indole and serine was investigated in a micellar solution of the surfactant Brij 56 in cyclohexane. An anion exchanger was employed to facilitate the transfer of tryptophan and serine between the water pool of the reverse micelle and the bulk organic phase. The influence of potassium ion, water content, pH, and co-surfactant on enzyme activity is reported. Kinetic studies indicate that the enzyme is not inhibited by indole in the micellar system and that the enzyme is more stable in reverse micelles than in bulk water. The design of a continuous reverse micelle reactor, which accommodates both product recovery and enzyme reactivation, is discussed. 相似文献
12.
The biphenyl-degrading Gram-negative bacterium Cupriavidus basilensis (formerly Ralstonia sp.) SBUG 290 uses various aromatic compounds as carbon and energy sources and has a high capacity to transform bisphenol A (BPA), which is a hormonally active substance structurally related to biphenyl. Biphenyl-grown cells initially hydroxylated BPA and converted it to four additional products by using three different transformation pathways: (a) formation of multiple hydroxylated BPA, (b) ring fission, and (c) transamination followed by acetylation or dimerization. Products of the ring fission pathway were non-toxic and all five products exhibited a significantly reduced estrogenic activity compared to BPA. Cell cultivation with phenol and especially in nutrient broth (NB) resulted in a reduced biotransformation rate and lower product quantities, and NB-grown cells did not produce all five products in detectable amounts. Thus, the question arose whether enzymes of the biphenyl degradation pathway are involved in the transformation of BPA and was addressed by proteomic analyses. 相似文献
13.
Sphingolipid ceramide N-deacylase catalyzes a reversible reaction in which the amide linkages of the ceramides of various sphingolipids are cleaved or synthesized. Hydrolysis of sphingolipids by the enzyme proceeded efficiently at acidic pH in the presence of high concentrations of detergents, whereas the reverse reaction tended to be favored at neutral pH with a decrease in the detergent concentration. Although the catalytic efficiency (V(max)/K(m)) of the hydrolysis and reverse reactions was changed mainly by the concentration of detergents in the reaction mixture, V(max) and K(m) for the reverse reaction were relatively higher than those for the forward reaction, irrespective of the detergent concentration. The reverse reaction proceeded most efficiently when the molar ratio of lyso-sphingolipids and fatty acids was fixed at 1 : 1-2, the yield of the reaction exceeding 70-80%. The reverse and exchange (transacylation) reactions did not require ATP, CoA, metal ions or addition of organic solvents. Studies using inhibitors and chemical modifiers of the enzyme protein suggested that both the hydrolysis and condensation reactions are catalyzed at the same catalytic domain. These results indicate that the reverse hydrolysis reaction of the enzyme is unique, being completely different from those of lipases, proteases and glycosidases reported to date. 相似文献
14.
This study developed a simple, efficient method for producing racemic β-phenylalanine acid (BPA) and its derivatives via the enantioselective acylation catalyzed by the penicillin G acylase from Alcaligenes faecalis (Af-PGA). When the reaction was run at 25°C and pH 10 in an aqueous medium containing phenylacetamide and BPA in a molar ratio of 2:1, 8 U/mL enzyme and 0.1 M BPA, the maximum BPA conversion efficiency at 40 min only reached 36.1%, which, however, increased to 42.9% as the pH value and the molar ratio of phenylacetamide to BPA were elevated to 11 and 3:1, respectively. Under the relatively optimum reaction conditions, the maximum conversion efficiencies of BPA derivatives all reached about 50% in a relatively short reaction time (45–90 min). The enantiomeric excess value of product ( eep ) and enantiomeric excess value of substrate ( ees ) were all above 98% and 95%, respectively. These results suggest that the method established in this study is practical, effective, and environmentally benign and may be applied to industrial production of enantiomerically pure BPA and its derivatives. 相似文献
15.
AbstractThe main objective of this study is the evaluation of the capability of laccase from Myceliophthora thermophila immobilized on fumed silica microparticles (fsMP) for the removal of endocrine disrupting chemicals (EDCs) in two enzymatic reactor configurations. This type of support can also be magnetized to allow the straightforward separation of the biocatalyst under a magnetic field. The support exhibited excellent biocompatibility with the enzyme, superior tolerance to pH and temperature as well as improved stability in comparison with the free enzyme, even in the presence of organic solvents and enzyme inhibitors. The technical feasibility of the removal of EDCs by immobilized laccase was assessed in two types of enzymatic reactors operated in sequential mode: a membrane reactor using fsMP-laccase and a reactor with magnetic separation using magnetized fsMP-laccase. The extent of transformation for the target compounds: bisphenol A (BPA) and 17β-estradiol (E2) was high and comparable to free laccase in both systems (up to 80%). The possibility of reusing the immobilized enzyme, especially for magnetized supports, offers an interesting approach in the development of enzyme based processes for the biotransformation of emerging pollutants. 相似文献
16.
Degradation of 2,2-Bis(4-hydroxyphenyl)propane (bisphenol A, BPA), an endocrine-disturbing chemical, by the growing mycelia of the white-rot basidiomycete, Pleurotus ostreatus, was examined. About 80% of BPA initially present decreased in 12 days of culture with this fungus. By in vitro experiments using the lignin-degrading enzyme manganese peroxidase (MnP), BPA was metabolized to phenol, 4-isopropenylphenol, 4-isopropylphenol, and hexestrol. The degradation products of BPA were assumed to be formed by the one-electron oxidation of the substrate. 相似文献
17.
The ability of certain organophosphorus (OP) compounds to inhibit acetylcholinesterase (AChE) has made them useful for industrial (insecticides) and military (nerve agents) purposes. We have previously published a single compartment mathematical model of the interactions between OP nerve agents and the enzymes affected by these agents. That model, which could be used to predict the LD50 of seven nerve agents in rats, has been extended to include the protective actions of stoichiometric and catalytic OP-scavenger enzymes (delivered as pretreatments) so that protective ratios attributable to the scavengers may be predicted. Prediction of expected human protection from in vitro rate constant and initial enzyme level measurements is the ultimate goal for this work. The enhanced model predicts the LD50 from rate constants of the OP agent's binding reactions with AChE, carboxylesterase (CaE) and a stoichiometric scavenger (S); a first-order OP elimination rate (including a contribution due to a catalytic scavenger); and whole body estimates of AChE, CaE and S. The ratio of the scavenger-treated LD50 estimate to the scavenger-free LD50 estimate provided a theoretical expression describing the scavenger's contributions to the protective ratio. Published in vivo protective ratios for two stoichiometric scavengers (fetal bovine serum AChE and human utyrylcholinesterase) against challenge by several OP agents in mice were compared with ratios predicted by the model. A linear regression analysis of in vivo protective ratios in mice versus the ratios predicted by the model from the in vitro measurements resulted in an R(2) value of 0.902. The catalytic scavenger portion of the theory could not be validated due to a lack of published data. We conclude that the one-compartment model can be used to make reasonable estimates of the protective ratio attributable to stoichiometric scavengers, but can make no conclusions regarding the ability of the model to predict catalytic scavenger protection ratios. 相似文献
18.
The enzymatic conversion of cholesterol to cholestenone by cholesterol oxidase (Brevibacterium sp.)in reversed micelles in a system composed of AOT/isooctane/water/cholesterol has been examined. The catalytic activity of the enzyme was correlated with the physicochemical properties of water in water-in-oil (w/o) microemulsion systems. In a system consisting of 3 wt % AOT in isooctane, reversed micelles started to form as the [H(2)O]/[AOT] (e.g., the w(0)) ratio increased above 4-5. The formation of reversed micelles with a core of neat (bulk) water was verified from determinations of both the partial molar volume of water and the scissors vibration of water [with Fourier transform infrared (FTIR) spectroscopy] in the w/o microemulsion systems. A plot of enzyme activity vs. w(0) indicated that the hydration of enzyme molecules per se was not sufficient to give rise to catalytic activity. Instead, it appeared that the formation of an aqueous micellar core was necessary for full activation of the enzyme. Based on micelle size distribution analysis, it was estimated that about one micelle per one thousand contained an enzyme molecule. Since the apparent reaction rate could be markedly enhanced by increasing the enzyme/water ratio, we conclude that the number of enzyme-containing micelles was an important rate-limiting factor in the system. 相似文献
19.
Inducible pyrrole-2-carboxylate decarboxylase from Bacillus megaterium PYR2910 catalyzes the decarboxylation of pyrrole-2-carboxylate to stoichiometric amounts of pyrrole and CO 2. A unique feature of the homodimeric enzyme is its requirement for an organic acid such as acetate, propionate, butyrate or pimelate. A catalytic mechanism including a cofactor function of the organic acid was proposed. Due to an equilibrium constant of 0.3–0.4 M, the enzyme also catalyzes the reverse carboxylation of pyrrole after the addition of bicarbonate. For the synthesis of pyrrole-2-carboxylate, the reverse reaction was optimized and the equilibrium shifted towards the carboxylate. The product yield was 230 mM (25.5 g/l) pyrrole-2-carboxylate from 300 mM pyrrole in a batch reaction and 325 mM (36.1 g/l) from 400 mM pyrrole in a fed batch reaction, using both whole cells and the purified enzyme in a pH 8.0 reaction mixture with bicarbonate saturation of 1.9 M. 相似文献
20.
BackgroundMale reproductive toxicity induced by exposure to bisphenol A (BPA) has been widely reported. The testes have proven to be a major target organ of BPA toxicity, so studying testicular metabolite variation holds promise for the discovery of mechanisms linked to the toxic effects of BPA on reproduction. Methodology/Principal FindingsMale Sprague-Dawley rats were orally administered doses of BPA at the levels of 0, 50 mg/kg/d for 8 weeks. We used an unbiased liquid chromatography-quadrupole time-of-flight (LC-QTOF)-based metabolomics approach to discover, identify, and analyze the variation of testicular metabolites. Two n-6 fatty acids, linoleic acid (LA) and arachidonic acid (AA) were identified as potential testicular biomarkers. Decreased levels of LA and increased levels of AA as well as AA/LA ratio were observed in the testes of the exposed group. According to these suggestions, testicular antioxidant enzyme levels were detected. Testicular superoxide dismutase (SOD) declined significantly in the exposed group compared with that in the non-exposed group, and the glutathione peroxidase (GSH-Px) as well as catalase (CAT) also showed a decreasing trend in BPA treated group. Conclusions/SignificanceBPA caused testicular n-6 fatty acid composition variation and decreased antioxidant enzyme levels. This study emphasizes that metabolomics brings the promise of biomarkers identification for the discovery of mechanisms underlying reproductive toxicity. 相似文献
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