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1.
Summary Two -glucosidase genes, designatedbglA andbglB, were isolated from a gene bank ofClostridium
thermocellum DSM 1237. The coding sequences forbglA andbglB were located on non-homologous DNA fragments of 3.2– and 3.4-kb, respectively. Both genes direct inEscherichia
coli the synthesis of cytoplasmic -glucosidases, which differ with respect to substrate specificity and temperature profile. The properties of thebglA-encoded -glucosidase A closely resemble that of a -glucosidase previously isolated fromC.
thermocellum cultures. 相似文献
2.
Min Lu Per- Ake Albertsson G te Johansson Folke Tjerneld 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1996,680(1-2):65-70
Benzoyl dextran with a degree of substitution of 0.18 was synthesized by reacting dextran T500 with benzoyl chloride. A new type of aqueous two-phase system composed of benzoyl dextran as bottom phase polymer and the random copolymer of ethylene oxide and propylene oxide (Ucon 50-HB-5100) as top phase polymer has been formed. The phase diagram for the system Ucon 50-HB-5100-benzoyl dextran with a degree of substitution of 0.18 was determined at room temperature. This two-phase system has been used to purify 3-phosphoglycerate kinase from bakers' yeast. The top-phase polymer (Ucon) can be separated from target enzyme by increasing the temperature. The bottom-phase polymer (benzyol dextran) could be recovered by addition of salt. Yeast homogenate was partitioned in a primary Ucon 50-HB-5100-benzoyl dextran aqueous two-phase system. After phase separation the top phase was removed and temperature-induced phase separation was used for formation of a water phase and a Ucon-rich phase. The benzoyl dextran-enriched bottom phase from the primary system was diluted, and the polymer was separated from water by addition of Na2SO4. 相似文献
3.
Pronounced inhibition of noradrenaline uptake by 10-hydroxymetabolites of nortriptyline 总被引:1,自引:0,他引:1
The potency of the tricyclic antidepressant nortriptyline (NT) relative to that of its major metabolite (E-10-hydroxynortriptyline; E-10-OH-NT) to inhibit the uptake of noradrenaline in rat brain slices incubated in human plasma was 1.75 ± S.D. 0.30. The E- and Z-isomers of 10-OH-NT were equipotent. Hydroxy-metabolites of NT or amitriptyline did not inhibited the neuronal uptake of serotonin.During treatment of 87 patients with NT or amitriptyline the mean ratio between the plasma levels of unconjugated 10-OH-NT and NT was 1.40 ± S.D. 0.86 with a range of 0.32 – 5.0. This ratio increased with age. There was a significant correlation between the plasma levels of the two compounds (r = 0.63; p < 0.001). In three patients treated with NT, the CSF levels of the parent drug and unconjugated 10-OH-NT were similar and about 5 % of the plasma levels.These results indicate that during treatment with NT (or amitriptyline) 10-OH-NT contributes to the effect of these drugs on central noradrenergic neurons. This seems to be of special significance in the elderly. 相似文献
4.
David I.C. Scopes Ewe Zarnack Nelson J. Leonard Ruth Y. Scmith Folke Skoog 《Phytochemistry》1976,15(10):1523-1526
We have tested the possible genesis of kinetin from a 2′-deoxyadenylate unit of DNA by a chemical route involving a head-to-tail transfer of deoxyribose from the 9 to the 3 position of the adenine nucleus via a cyclonucleoside, with subsequent elimination of 1′- and 3′-polar groups and 3 → N6 intramolecular rearrangement leading to kinetin. We have also determined quantitatively the per cent conversions to 3-furfuryladenine and/or kinetin of the following under autoclaving conditions at 120°, pH 4, 2 atm, and 4 hr: (1) adenine/furfury alcohol; (2) adenine/2-deoxy-d-ribose; (3) 2′-deoxyadenosine; (4) 3-furfuryladenine; (5) 3,5′-(3′-O-diethylphosphoryl-2′-deoxya-denosine)-cyclonucleoside p-toluenesulfonate. The sequence of reactions involving cyclonucleoside formation and rearrangement has been shown to be a chemically feasible route by which kinetin can be formed, although it is not the only way this cytokinin can be generated. 相似文献
5.
6.
Qingling Zhang Michael Cox Zhike Liang Folke Brinkmann Paul A Cardenas Rachael Duff Pankaj Bhavsar William Cookson Miriam Moffatt Kian Fan Chung 《PloS one》2016,11(4)
BackgroundThe lower airways harbor a community of bacterial species which is altered in asthma.ObjectivesWe examined whether the lower airway microbiota were related to measures of asthma severity.MethodsWe prospectively recruited 26 severe asthma, 18 non-severe asthma and 12 healthy subjects. DNA was extracted from induced sputum and PCR amplification of the V3-V5 region of bacterial 16S rRNA gene was performed.ResultsWe obtained 138,218 high quality sequences which were rarefied at 133 sequences/sample. Twenty OTUs had sequences ≥1% of total. There were marked differences in the distribution of Phyla between groups (P = 2.8x10-118). Bacteroidetes and Fusobacteria were reduced in non-severe and severe asthmatic groups. Proteobacteria were more common in non-severe asthmatics compared to controls (OR = 2.26; 95% CI = 1.94–2.64) and Firmicutes were increased in severe asthmatics compared to controls (OR = 2.15; 95%CI = 1.89–2.45). Streptococcal OTUs amongst the Firmicutes were associated with recent onset asthma, rhinosinusitis and sputum eosinophilia.ConclusionsSputum microbiota in severe asthma differs from healthy controls and non-severe asthmatics, and is characterized by the presence of Streptococcus spp with eosinophilia. Whether these organisms are causative for the pathophysiology of asthma remains to be determined. 相似文献
7.
Dolby V Lundqvist A Fröberg T Lüllau E Shaw J Tjerneld F Cronet P 《Journal of biochemical and biophysical methods》2004,58(3):195-205
The G-protein coupled melanocortin 4 receptor (MC4r) plays an important role in the energy metabolism. We overexpressed the MC4r in CHO cells and performed characterisation studies on the cell membranes to determine functional stability and ligand binding properties of the receptor. The affinity for the ligands [Nle4, d-Phe7]-alphaMSH and MTII was lost below pH 6 but could be restored by returning to physiological pH. Increasing NaCl concentration up to 1 M had little influence on the binding of either ligand. At neutral pH, physiological salt concentration and 4 degrees C the ligand affinity of the receptor was stable for up to 6 days. These findings will facilitate design of purification methods for the receptor. 相似文献
8.
The presence of lignin has shown to play an important role in the enzymatic degradation of softwood. The adsorption of enzymes, and their constituent functional domains on the lignocellulosic material is of key importance to fundamental knowledge of enzymatic hydrolysis. In this study, we compared the adsorption of two purified cellulases from Trichoderma reesei, CBH I (Cel7A) and EG II (Cel5A) and their catalytic domains on steam pretreated softwood (SPS) and lignin using tritium labeled enzymes. Both CBH I and its catalytic domain exhibited a higher affinity to SPS than EG II or its catalytic domain. Removal of cellulose binding domain decreased markedly the binding efficiency. Significant amounts of CBH I and EG II also bound to isolated lignin. Surprisingly, the catalytic domains of the two enzymes of T. reesei differed essentially in the adsorption to isolated lignin. The catalytic domain of EG II was able to adsorb to alkaline isolated lignin with a high affinity, whereas the catalytic domain of CBH I did not adsorb to any of the lignins tested. The results indicate that the cellulose binding domain has a significant role in the unspecific binding of cellulases to lignin. 相似文献
9.
Melatonin induces oscillations in the peroxidase-oxidase (PO) reaction catalyzed by horseradish peroxidase. We present here studies of the effect of pH, enzyme concentration, and concentration of melatonin on the oscillation frequency. We also present a mechanistic model to explain the experimentally observed changes in oscillation frequency. Using the data obtained here we are able to predict that oscillations will also occur in the PO reaction catalyzed by myeloperoxidase. Myeloperoxidase is an important protein in activated neutrophils and we provide evidence that the oscillations of NAD(P)H, superoxide and hydrogen peroxide in these cells may involve this enzyme. Thus, our experimental system can be considered a model system for the nonrespiratory oxygen metabolism in activated neutrophils and other similar cells participating in the defence against invading pathogens. 相似文献
10.
Berggren K Wolf A Asenjo JA Andrews BA Tjerneld F 《Biochimica et biophysica acta》2002,1596(2):253-268
It is of great interest and importance how different amino acid residues contribute to and affect the properties of a protein surface. Partitioning in aqueous two-phase systems has the potential to be used as a rapid and simple method for studying the surface properties of proteins. The influence on partitioning of the surface exposed amino acid residues of eight structurally determined monomeric proteins has been studied. The proteins were characterized in terms of surface exposed residues with a computer program, Graphical Representation and Analysis of Surface Properties (GRASP), and partitioned in two EO30PO70-dextran aqueous two-phase systems, only differing in polymer concentrations (system I: 6.8% EO30PO70, 7.1% dextran; system II: 9% EO30PO70, 9% dextran). We show for the first time that the partitioning behaviour of different monomeric proteins can be described by the differences in surface exposed amino acid residues. The contribution to the partition coefficient of the residues was found to be best characterized by peptide partitioning in the aqueous two-phase system. Compared to hydrophobicity scales available in the literature, each amino acid contribution is characterized by the slope given by the graph of log K against peptide chain length, for peptides of different length containing only one kind of residue. It was also shown that each amino acid contribution is relative to the total protein surface and the other residues on the surface. Surface hydrophobicity calculations realized for systems I and II gave respectively correlation coefficients of 0.961 and 0.949 for the linear relation between log K and calculated hydrophobicity values. To study the effect on the partition coefficient of different amino acids, they were grouped into classes according to common characteristics: the presence of an aromatic group, a long aliphatic chain or the presence of charge. Using these groups it was possible to confirm that aromatic residues have the strongest effect on the partition coefficient, giving preference to the upper EO30PO70 phase of the system; on the other hand the presence of charged amino acids on the protein surface enhances the partition of the protein to the lower dextran phase. It is also important to note that the sensitivity of the EO30PO70-dextran system for the surface exposed residues was increased by increasing the polymer concentrations. The partition coefficient of a monomeric protein can thus be predicted from its surface exposed amino acid residues and the system can also be used to characterize protein surfaces of monomeric proteins in general. 相似文献