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61.
Björn V. Adalbjörnsson Helen S. Toogood Dr. Anna Fryszkowska Dr. Christopher R. Pudney Dr. Thomas A. Jowitt Dr. David Leys Dr. Nigel S. Scrutton Prof. 《Chembiochem : a European journal of chemical biology》2010,11(2):197-207
We report the crystal structure of a thermophilic “ene” reductase (TOYE) isolated from Thermoanaerobacter pseudethanolicus E39. The crystal structure reveals a tetrameric enzyme and an active site that is relatively large compared to most other structurally determined and related Old Yellow Enzymes. The enzyme adopts higher order oligomeric states (octamers and dodecamers) in solution, as revealed by sedimentation velocity and multiangle laser light scattering. Bead modelling indicates that the solution structure is consistent with the basic tetrameric structure observed in crystallographic studies and electron microscopy. TOYE is stable at high temperatures (Tm>70 °C) and shows increased resistance to denaturation in water‐miscible organic solvents compared to the mesophilic Old Yellow Enzyme family member, pentaerythritol tetranitrate reductase. TOYE has typical ene‐reductase properties of the Old Yellow Enzyme family. There is currently major interest in using Old Yellow Enzyme family members in the preparative biocatalysis of a number of activated alkenes. The increased stability of TOYE in organic solvents is advantageous for biotransformations in which water‐miscible organic solvents and biphasic reaction conditions are required to both deliver novel substrates and minimize product racemisation. 相似文献
62.
Geoffray Labar Cédric Bauvois Dr. Franck Borel Dr. Jean‐Luc Ferrer Dr. Johan Wouters Prof. Didier M. Lambert Prof. 《Chembiochem : a European journal of chemical biology》2010,11(2):218-227
2‐Arachidonoylglycerol plays a major role in endocannabinoid signaling, and is tightly regulated by the monoacylglycerol lipase (MAGL). Here we report the crystal structure of human MAGL. The protein crystallizes as a dimer, and despite structural homologies to haloperoxidases and esterases, it distinguishes itself by a wide and hydrophobic access to the catalytic site. An apolar helix covering the active site also gives structural insight into the amphitropic character of MAGL, and likely explains how MAGL interacts with membranes to recruit its substrate. Docking of 2‐arachidonoylglycerol highlights a hydrophobic and a hydrophilic cavity that accommodate the lipid into the catalytic site. Moreover, we identified Cys201 as the crucial residue in MAGL inhibition by N‐arachidonylmaleimide, a sulfhydryl‐reactive compound. Beside the advance in the knowledge of endocannabinoids degradation routes, the structure of MAGL paves the way for future medicinal chemistry works aimed at the design of new drugs exploiting 2‐arachidonoylglycerol transmission. 相似文献
63.
Jing Jin Sunil Chhatre Nigel J. Titchener‐Hooker Daniel G. Bracewell 《Journal of chemical technology and biotechnology (Oxford, Oxfordshire : 1986)》2010,85(2):209-215
BACKGROUND: Production of recombinant virus‐like particles (VLPs) in yeast expression systems for use as vaccines requires cell disruption and detergent‐mediated steps to liberate the product. Typically, these release high levels of cellular components such as lipids that foul chromatography columns. This study compares the impact of applying lipid‐rich and lipid‐depleted feedstocks to hydrophobic interaction chromatography columns to quantify the loss of performance caused by the presence of host lipids over a total of 40 operational cycles. RESULTS: VLP binding capacity in the lipid‐rich feed was significantly lower than for the lipid‐depleted feed, with greater than 24% of the lipids remaining in the column after each cycle. Triacylglycerol was found to be the major contaminant. The effectiveness of subsequent caustic clean‐in‐place was limited, resulting in column hydrophobicity increasing over repeated loading cycles. This improved the effective VLP binding capacity and affinity, but also made product elution more difficult, and recovery decreased by more than 70% over the 40 cycles. CONCLUSION: Host cell lipids cause major fouling problems during VLP purification. Instead of screening for better CIP conditions, priority should be given to identifying suitable upstream lipid removal strategies in order to maintain column performance and so yield more economically viable processes. Copyright © 2009 Society of Chemical Industry 相似文献
64.
Xiangdong Su Dr. Fabienne Pradaux‐Caggiano Dr. Mark P. Thomas Dr. Michelle W. Y. Szeto Dr. Heather A. Halem Dr. Michael D. Culler Dr. Nigel Vicker Dr. Barry V. L. Potter Prof. 《ChemMedChem》2010,5(7):1026-1044
11β‐Hydroxysteroid dehydrogenases (11β‐HSDs) are key enzymes regulating the pre‐receptor metabolism of glucocorticoid hormones. The modulation of 11β‐HSD type 1 activity with selective inhibitors has beneficial effects on various conditions including insulin resistance, dyslipidemia and obesity. Inhibition of tissue‐specific glucocorticoid action by regulating 11β‐HSD1 constitutes a promising treatment for metabolic and cardiovascular diseases. A series of novel adamantyl ethanone compounds was identified as potent inhibitors of human 11β‐HSD1. The most active compounds identified ( 52 , 62 , 72 , 92 , 103 and 104 ) display potent inhibition of 11β‐HSD1 with IC50 values in the 50–70 nM range. Compound 72 also proved to be metabolically stable when incubated with human liver microsomes. Furthermore, compound 72 showed very weak inhibitory activity for human cytochrome P450 enzymes and is therefore a candidate for in vivo studies. Comparison of the publicly available X‐ray crystal structures of human 11β‐HSD1 led to docking studies of the potent compounds, revealing how these molecules may interact with the enzyme and cofactor. 相似文献
65.
Ruth U Barbante C Bigler M Delmonte B Fischer H Gabrielli P Gaspari V Kaufmann P Lambert F Maggi V Marino F Petit JR Udisti R Wagenbach D Wegner A Wolff EW 《Environmental science & technology》2008,42(15):5675-5681
To improve quantitative interpretation of ice core aeolian dust records, a systematic methodological comparison was made. This involved methods for water-insoluble particle counting (Coulter counter and laser-sensing particle detector), soluble ion analysis (ion chromatography and continuous flow analysis), elemental analysis (inductively coupled plasma mass spectroscopy at pH 1 and after full acid digestion), and water-insoluble elemental analysis (proton induced X-ray emission). Antarctic ice core samples covering the last deglaciation from the EPICA Dome C (EDC) and the EPICA Dronning Maud Land (EDML) cores were used. All methods correlate very well among each other, but the ratios of glacial age to Holocene concentrations, which are typically a factor approximately 100, differ between the methods by up to a factor of 2 with insoluble particles showing the largest variability. The recovery of ICP-MS measurements depends on the digestion method and is differentfor different elements and during different climatic periods. EDC and EDML samples have similar dust composition, which suggests a common dust source or a common mixture of sources for the two sites. The analyzed samples further reveal a change of dust composition during the last deglaciation. 相似文献
66.
An explanation for the effect of inoculum size on MIC and the growth/no growth interface 总被引:1,自引:0,他引:1
The inoculum effect (IE) is the phenomenon observed where changes in the inoculum size used in an experiment alters the outcome with respect to, for example, the minimum inhibitory concentration of an antimicrobial or the growth/no growth boundary for a given set of environmental conditions. Various hypotheses exist as to the cause of the IE such as population heterogeneity and quorum sensing, as well as the null hypothesis - that it is artefactual. Time to detection experiments (TTD) were carried out on different initial inoculum sizes of several bacterial species (Aeromonas hydrophila, Enterobacter sakazakii, Salmonella Poona, Escherichia coli and Listeria innocua) when challenged with different pH and with combined pH and sodium acetate. Data were modelled using a modification to a Gamma model (Lambert and Bidlas 2007, Int. J. Food Microbiology 115, 204-213), taking into account the inoculum size dependency on the TTD obtained under ideal conditions. The model suggests that changes in minimum inhibitory concentration (MIC) or in the growth-no growth boundary with respect to inoculum size are due to using a smaller or larger inoculum (i.e. is directly related to microbial number) and is not due to other, suggested, phenomena. The model used further suggests that the effect of a changing inoculum size can be modelled independently of any other factor, which implies that a simple 1 to 2-day experiment measuring the TTD of various initial inocula can be used as an adjunct to currently available models. 相似文献
67.
Firmesse O Alvaro E Mogenet A Bresson JL Lemée R Le Ruyet P Bonhomme C Lambert D Andrieux C Doré J Corthier G Furet JP Rigottier-Gois L 《International journal of food microbiology》2008,125(2):176-181
The objective of this study was to determine i) if Camembert cheese micro-organisms could be detected in fecal samples after regular consumption by human subjects and ii) the consequence of this consumption on global metabolic activities of the host colonic microbiota. An open human protocol was designed where 12 healthy volunteers were included: a 2-week period of fermented products exclusion followed by a 4-weeks Camembert ingestion period where 2x40 g/day of Camembert cheese was consumed. Stools were collected from the volunteers before consumption, twice during the ingestion period (2nd and 4th week) and once after a wash out period of 2 weeks. During the consumption of Camembert cheese, high levels of Lactococcus lactis and Leuconostoc mesenteroides were measured in fecal samples using real-time quantitative PCR, reaching median values of 8.2 and 7.5 Log(10) genome equivalents/g of stool. For Ln. mesenteroides, persistence was observed 15 days after the end of Camembert consumption. The survival of Geotrichum candidum was also assessed and the fecal concentration reached a median level of 7.1 Log(10) CFU/g in stools. Except a decreasing trend of the nitrate reductase activity, no significant modification was shown in the metabolic activities during this study. 相似文献
68.
Jennifer O'Loughlin Marie Lambert Igor Karp Jennifer McGrath Katherine Gray-Donald Tracie A Barnett Edgard E Delvin Emile Levy Gilles Paradis 《Nicotine & tobacco research》2008,10(3):525-532
Although related to inflammatory markers in adults, little is known about the association between cigarette smoking and C-reactive protein (CRP) in adolescent smokers. We examined the association between high-sensitivity CRP (hs-CRP) concentrations and smoking in youth. We used data from a cross-sectional, province-wide survey of a representative sample of youth conducted in Quebec, Canada, in 1999. Data were collected in self-report questionnaires completed by participants and their parents. Participants provided a fasting blood sample, and anthropometric measures were undertaken by trained technicians. The present analysis pertains to 1,501 adolescents aged 13 and 16 years who completed questionnaires and for whom blood samples were available. The independent association between a six-category indicator of smoking status and elevated hs-CRP, defined as a value at least in the 90th percentile of the age- and sex-specific CRP distribution, was assessed in multiple logistic regression analyses controlling for potential confounders. Relative to never-smokers, the odds ratios (95% confidence intervals) for puffers (i.e., never smoked a whole cigarette), those who smoked but not in the past month, light past-month smokers, moderate past-month smokers, and heavy past-month smokers were 1.04 (0.55-1.98), 1.76 (1.06-2.94), 1.39 (0.70-2.76), 2.07 (0.96-4.42), and 2.40 (1.18-4.88), respectively. Our data suggest a positive association between smoking status and elevated CRP in adolescents, and in particular among heavier past-month smokers. Damage related to cigarette smoking may begin soon after tobacco use initiation, reinforcing the preventive message that no level of smoking is safe in youth. 相似文献
69.
Adaptation of urban water supply infrastructure to impacts from climate and socioeconomic changes: The case of Hamilton,New Zealand 总被引:1,自引:4,他引:1
Matthias Ruth Clark Bernier Nigel Jollands Nancy Golubiewski 《Water Resources Management》2007,21(6):1031-1045
While the relations between climate variables and sectoral water demand have been well established in the literature, few
studies have attempted to quantify changes in urban water usage with climate change. Concentrating on the city of Hamilton,
New Zealand, we investigate possible water use and infrastructure needs for a range of climate and population projections.
We find that water demand (at the monthly aggregate level) is largely driven by changes in population, and not significantly
affected by changes in climate. However, as population increases, the effect of climate variables on per capita consumption
will be magnified. Monthly aggregate changes may further mask potenially significant short-term shortages. In several scenarios,
water supply shortages in 2030 occur with a 30--40% probability, suggesting needs for long-term capacity expansion or aggressive
demand side management, rather than implementation of short-term management of water demand. 相似文献
70.
Bueno M Astruc A Lambert J Astruc M Behra P 《Environmental science & technology》2001,35(7):1411-1419
Tributyltin (TBT) is the most important organotin compound that has been introduced into aquatic ecosystems. A better understanding of its interactions with solid surfaces is essential to estimate the possibilities of TBT migration through subsurface environments. For this purpose, TBT sorption onto a porous matrix of natural origin, a quartz sand as an aquifer material, was studied at low concentration levels with a monodirectional model of column type allowing sequential investigation of sorption and desorption processes. Different treatments of the solid phase were performed by injecting alkaline solutions, NaOH at pH 10.8 or NaClO-NaCl at pH 11.5, by decreasing the ionic strength or by adding kaolinite to change the surface composition and properties. The removal of iron and aluminum (hydr)oxides from the sand surface did not affect so much the sorption (decrease in 14% as compared to sorption on the raw sand). The original use of X-ray photoelectron spectroscopy to control treatment efficiency and to characterize sand surface modifications permitted to relate TBT sorption onto the aquifer material to quartz, the main component of the sand, and clay minerals (mainly kaolinite) present at trace levels at the sand surface. A first attempt of transport modeling with these two surface sites showed the consistency of our assumption. Moreover, estimation of Langmuir-type constants showed that TBT sorption affinity for the quartz surface (KL = 26.7 L micromol(-1)) was much greater than for kaolinite (KL = 6.3 L micromol(-1)). 相似文献