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991.
Juan M. Bolivar Tanja Consolati Torsten Mayr Bernd Nidetzky 《Biotechnology and bioengineering》2013,110(8):2086-2095
Enzymatic O2‐dependent oxidations are receiving increased attention for use in fine chemicals synthesis. Solid supported oxidation catalysts often show poor efficiency due to pronounced O2 diffusion restriction. Internal O2 supply therefore constitutes a key parameter for optimizing the enzyme immobilization. We herein describe an optical sensing method for quantitation of space‐averaged intraparticle O2 concentrations in porous Sepabeads carriers. The method applies phosphorescence lifetime measurements on Sepabeads labeled with an O2 sensitive indicator dye. Using glucose oxidase immobilized at different loadings (0.005–12 mg/g) on labeled Sepabeads, we analyzed in real time during the enzymatic reaction the formation of O2 concentration differences between bulk liquid and the intraparticle environment. We show that the O2 gradient at apparent steady state increased with increasing enzyme loading, so that O2 eventually became totally depleted from inside the highly loaded carriers. We also show that the residual intraparticle O2 concentration was correlated with the catalytic effectiveness factor (η) of the enzyme immobilizate used, thus providing a direct measure of the magnitude of O2 diffusion limitation. Once corrected for diffusional effect, η was no longer dependent on enzyme loading and its constant value now described the intrinsic activity of immobilized glucose oxidase. Three common procedures of enzyme immobilization, involving adsorption, cross‐linking, and covalent attachment, are shown to differ widely concerning the obtained intrinsic activity. Therefore, intraparticle O2 concentration data enable distinction between diffusional restriction and activity loss as the two principal factors limiting the effectiveness of immobilized O2 dependent enzymes, and thus they inform rational design of an optimally active oxidation biocatalyst on solid support. Biotechnol. Bioeng. 2013; 110: 2086–2095. © 2013 Wiley Periodicals, Inc. 相似文献
992.
993.
Torsten Gunnarsson Anders Karlsson Per Hansson Göran Johnson Christer Alling Göran Odham 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1998,705(2):1428
The `pathologic' phospholipid, phosphatidylethanol (PEth), formed only in the presence of ethanol, was determined in extracts of human blood using high-performance liquid chromatography with evaporative light scattering detection (ELSD) or electrospray (ES) mass spectrometry. Separation was performed using a diol column and a normal-phase binary gradient system. Decreasing concentrations of PEth (15 to 1 nmol/ml blood) could be detected by ELSD in three male alcoholics, up to 3 weeks after the beginning of an alcohol-free period. Using ES, levels down to 100 pmol/ml blood was detected. The molecular species of PEth were similar to those of phosphatidylcholine found in the same blood sample. The method provides a rapid quantitative and qualitative determination of PEth in blood. The limits of detection were 200 pmol (≈125 ng) using ELSD and 140 fmol (≈100 pg) using ES, total amounts injected. 相似文献
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995.
996.
Following a study of the relationship between cytokinin oxidase/dehydrogenase (CKX) and senescence in darkened barley leaf segments, we have now investigated the influence of light on the in vitro activity of CKX. Seedlings of Hordeum vulgare L. were grown for 8 d under a light/dark regime of 18 h white light and 6 h darkness. Then apical parts of 7 cm length were cut from the first foliage leaves and their bases were placed in water. In segments kept in the dark, the CKX activity measured by cleavage of N6-(Δ2-isopentenyl)adenine rose from 0.1 pkat (g FW)−1 to 0.8 pkat (g initial FW)−1 within the first 4 d of incubation. In contrast, in segments kept under the light/dark regime it reached a value of 8.6 pkat (g initial FW)−1 over the same time period. The chlorophyll a content declined slightly slower during light/dark cycling than in darkness. In contrast to segments and isolated laminae, corresponding attached laminae exhibited less CKX activity after 2 d under light/dark conditions than after 2 d in the dark. The activity in attached laminae of first foliage leaves of plants growing in light/dark cycling increased strongly only when the plants were older than 4 weeks. In line with this, the CKX activity in attached laminae of flag leaves of barley growing in fields increased in a late developmental state. The senescence of darkened isolated laminae of Zea mays L. and Phragmites australis (Cav.) Trin. ex Steudel was associated with an enhancement of CKX activity too. Because in most cases a positive correlation between CKX activity and senescence was found, it is likely that the enzyme promotes senescence by destroying cytokinins, which help to keep Poaceae leaves green. Light may promote not only cytokinin degradation but also the formation of bioactive cytokinins in leaf segments. 相似文献
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998.
Although sessile, plants are able to grow toward or away from an environmental stimulus. Important examples are stem or leaf orientation of higher plants in response to the direction of the incident light. The responsible photoreceptors belong to the phototropin photoreceptor family. Although the mode of phototropin action is quite well understood, much less is known of how the light signal is transformed into a bending response. Several lines of evidence indicate that a lateral auxin gradient is responsible for asymmetric cell elongation along the light gradient within the stem. However, some of the molecular key players leading to this asymmetric auxin distribution are, as yet, unidentified. Previously, it was shown that phototropin gets autophosphorylated upon illumination and binds to a scaffold protein termed NPH3 (for nonphototropic hypocotyl 3). Using a yeast three-hybrid approach with phototropin and NPH3 as a bait complex, we isolated a protein, termed EHB1 (for enhanced bending 1), with a so far unknown function, which binds to this binary complex. This novel interacting factor negatively affects hypocotyl bending under blue light conditions in Arabidopsis (Arabidopsis thaliana) and thus seems to be an important component regulating phototropism. Interestingly, it could be shown that the gravitropic response was also affected. Thus, it cannot be ruled out that this protein might also have a more general role in auxin-mediated bending toward an environmental stimulus. 相似文献
999.
Plant invasions can have substantial consequences for the soil ecosystem, altering microbial community structure and nutrient
cycling. However, relatively little is known about what drives these changes, making it difficult to predict the effects of
future invasions. In addition, because most studies compare soils from uninvaded areas to long-established dense invasions,
little is known about the temporal dependence of invasion impacts. We experimentally manipulated forest understory vegetation
in replicated sites dominated either by exotic Japanese barberry (Berberis thunbergii), native Viburnums, or native Vacciniums, so that each vegetation type was present in each site-type. We compared the short-term effect of vegetation changes to the
lingering legacy effects of the previous vegetation type by measuring soil microbial community structure (phospholipid fatty
acids) and function (extracellular enzymes and nitrogen mineralization). We also replaced the aboveground litter in half of
each plot with an inert substitute to determine if changes in the soil microbial community were driven by aboveground or belowground
plant inputs. We found that after 2 years, the microbial community structure and function was largely determined by the legacy
effect of the previous vegetation type, and was not affected by the current vegetation. Aboveground litter removal had only
weak effects, suggesting that changes in the soil microbial community and nutrient cycling were driven largely by belowground
processes. These results suggest that changes in the soil following either invasion or restoration do not occur quickly, but
rather exhibit long-lasting legacy effects from previous belowground plant inputs. 相似文献
1000.
Mukherjee K Abu Mraheil M Silva S Müller D Cemic F Hemberger J Hain T Vilcinskas A Chakraborty T 《Applied and environmental microbiology》2011,77(12):4237-4240
We report the use of antimicrobial hemolymph proteins from the model host Galleria mellonella as an inhibitor for various Listeria strains, providing a novel source for antilisterial therapeutics. We also have shown that specific virulence-associated genes known to mediate antimicrobial resistance of Listeria in mammalian models indicated a similar function in Galleria. 相似文献