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121.
Minoru Isobe Prof. Dr. Masakuni Kurono Dr. Katsunori Tsuboi Dr. Akira Takai Prof. Dr. 《化学:亚洲杂志》2007,2(3):377-385
We have accomplished the synthesis of 13C‐labeled tautomycin at the C18, C19, C21, and C22 positions starting from 100 % [13C]triethylphosphonoacetate for the purpose of elucidating the dynamics and conformation of the C17–C26 moiety. NMR spectroscopy of 13C‐labeled tautomycin revealed strong binding with protein phosphatase type 1 and new features in the 13C NMR spectrum, such as the very small three‐bond coupling constants (2J). 相似文献
122.
We studied thermal transitions and physical stability
of oil-in-water emulsions containing different milk fat compositions, arising
from anhydrous milk fat alone (AMF) or in mixture (2:1 mass ratio) with a
high melting temperature (AMF–HMT) or a low melting temperature (AMF–LMT)
fraction. Changes in thermal transitions in bulk fat and emulsion samples
were monitored by differential scanning calorimetry (DSC) under controlled
cooling and reheating cycles performed between 50 and –45°C (5°C
min–1). Comparison between bulk fat samples
and emulsions indicated similar values of melting completion temperature,
whereas initial temperature of fat crystallization (Tonset)
seemed to be differently affected by storage temperature depending on triacylglycerols
(TAG) composition. After storage at 4°C, Tonset
values were very similar for emulsified and non-emulsified AMF–HMT blend,
whereas they were lower (by approx. 6°C) for emulsions containing AMF
or mixture of AMF–LMT fraction. After storage at –30°C, Tonset values of re-crystallization
were higher in emulsion samples than in bulk fat blends, whatever the TAG
fat composition. Light scattering measurements and fluorescence microscopic
observations indicated differences in fat droplet aggregation-coalescence
under freeze-thaw procedure, depending on emulsion fat composition. It appeared
that under quiescent freezing, emulsion containing AMF–LMT fraction
was much less resistant to fat droplet aggregation-coalescence than emulsions
containing AMF or AMF–HMT fraction. Our results indicated the role of
fat droplet liquid-solid content on emulsion stability. 相似文献
123.
Thereza Christina Vessoni Penna Eb Chiarini Adalberto Pessoa Junior 《Applied biochemistry and biotechnology》2003,107(1-3):481-491
Transformed cells of Escherichia coli expressing recombinant green fluorescent protein (GFPuv) were subjected to two methods of extraction: (1) freezing/thawing/sonication
(FTS) cycles prior to the three-phase partitioning (TPP) method, or (2) directly to TPP extraction. The amount of GFPuv released
by the FTS plus TPP method varied: 374μg/mL (first cycle), 93–442 μg/mL (second cycle), 32–359 μg/mL (third cycle), 18–115
μg/mL (fourth cycle). The GFPuv yields by the second method (TPP only) were, 23–54 μg/mL for the first extract and 33–91 μg/mL
for the second. The FTS plus TPP method released similar amounts of GFPuv to that extracted by TPP; and provided a better
mixture elution through the hydrophobic interaction column: 13–63 μg/mL for FTS plus TPP methods, and 2.5–13 μg/mL for TPP.
The results showed that although selective permeation is a more laborious methodology, it was more efficient for obtaining
of GFPuv in relation to the direct extraction of the cells for TPP. 相似文献
124.
Stavroula Theodorolea 《Analytica chimica acta》2005,547(1):132-137
A method was developed for the determination of selenium in human milk using electrothermal atomic absorption spectrometry. The use of chemical modifiers as well as their implications during the pyrolysis step was examined. The chemical modifiers that were studied were Zr, Ir as well as the mixed modifier Zr-Ir. The Ir modifier stabilized selenium at 1000 °C, Zr at 800 °C, while the mixed modifier at 1200 °C. The effect of modifier mass was studied and was found that better results are achieved with addition of 2 μg Zr and 2 μg Ir. The characteristic masses of selenium in the presence of Zr, Ir and the mixed modifier were found to be 73.3, 18.0 and 14.7 pg, respectively, while the corresponding limits of detection were found 2.0, 0.50 and 0.41 μg l−1. Consequently better results were obtained with the mixed modifier. The developed method was applied for the determination of selenium in human milk, which was digested with a HNO3 + H2O2 mixture in a microwave oven. The limit of detection of the method was 1.37 μg l−1, the characteristic mass, m0, was 48.8 pg and the repeatability was less than 5% as R.S.D.(%). Matrix matched calibration was used. Recoveries were estimated to be 93-105%. The method was applied to breast milk of Greek women (n = 9) and the Se content was found to be in the range 16.7-42.6 μg l−1 with mean value 27.4 ± 5.5 μg l−1. 相似文献
125.
Fipronil, a phenylpyrazole insecticide introduced for pest control on a broad range of crops, can also affect non-target insects such as honeybees. More widely, non-target environment such as milk produced by dairy cows fed with maize silage from treated seeds (=silage T) can be affected. To assess the potential transfer of fipronil residues (sulfone, sulfide, fipronil, desulfinyl and amide), a methodology including gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) analysis was developed and validated according to the 2002/657/EC decision, in order to reach a level of quantification below 0.1 μg L−1 in milk and 0.1 μg kg−1 in plants. Twelve dairy cows were fed with silage T during 4 months. Concentration of fipronil in treated seeds was estimated at 1 g kg−1, whereas silage from these seeds contained 0.30 ± 0.05 μg kg−1 of dry material of fipronil, 0.13 ± 0.03 μg kg−1 of dry material of sulfone. Sulfide residues were below the limit of quantification. Silage from untreated seeds (=silage U) presented traces of fipronil and sulfone, respectively at 0.04 ± 0.06 and 0.02 ± 0.03 μg kg−1 of dry material. Contribution of fipronil residues from supplies was insignificant. During administration of silage T, only sulfone residues were quantified in milk. The average concentration was 0.14 ± 0.05 μg L−1. Before and after administration, sulfone residues were detected but not quantifiable (<0.025 μg L−1). Our results suggest a transfer of fipronil from feed to milk under its sulfone form. Moreover, traces of fipronil residues in maize U, soya, wheat and straw show a diffuse contamination of this pesticide in the environment. 相似文献
126.
127.
Khaja Basheeruddin Vicki Rothman Simeon Margolis 《Applied biochemistry and biotechnology》1985,11(2):133-140
We have immobilized E.coli alkaline phosphatase (EC 3.1.3.1) by linking it covalently to sepharose 4B. This preparation has several advantages over
the soluble enzyme. The immobilized enzyme is easily separable from other constituents in incubation mixtures. The immobilized
enzyme can be reused repeatedly and is more stable than the soluble enzyme to heat treatment in the presence of 10 mM Mg2+. The insoluble and soluble phosphatases removed 75 and77%, respectively, of the inorganic phosphorus from casein. The immobilized enzyme inactivated two enzymes believed to be active
in the phosphorylated state, acyl-CoA : cholesterol acyltransferase (ACAT) by 39% and NADPH-cytochrome P-450 reductase by
89%. The utility of immobilized alkaline phosphatase for studying the phosphorylation and dephosphorylation of soluble or
membrane-bound enzymes and proteins is discussed. 相似文献
128.
Recent experimental work carried out in this laboratory on the ultrafast dynamics of myoglobin (Mb) is summarized with a stress on structural and vibrational energy relaxation. Studies on the structural relaxation of Mb following CO photolysis revealed that the structural change of heme itself, caused by CO photodissociation, is completed within the instrumental response time of the time-resolved resonance Raman apparatus used (approximately 2 ps). In contrast, changes in the intensity and frequency of the iron-histidine (Fe-His) stretching mode upon dissociation of the trans ligand were found to occur in the picosecond regime. The Fe-His band is absent for the CO-bound form, and its appearance upon photodissociation was not instantaneous, in contrast with that observed in the vibrational modes of heme, suggesting appreciable time evolution of the Fe displacement from the heme plane. The band position of the Fe-His stretching mode changed with a time constant of about 100 ps, indicating that tertiary structural changes of the protein occurred in a 100-ps range. Temporal changes of the anti-Stokes Raman intensity of the v4 and v7 bands demonstrated immediate generation of vibrationally excited heme upon the photodissociation and decay of the excited populations, whose time constants were 1.1 +/- 0.6 and 1.9 +/- 0.6 ps, respectively. In addition, the development of the time-resolved resonance Raman apparatus and prospects in this research field are described. 相似文献
129.
J. L. Gmez-Ariza V. Bernal-Daza M. J. Villegas-Portero 《Analytica chimica acta》2004,520(1-2):229-235
A comparison of chiral separation and analysis of selenomethionine in breast and formula milk, using high performance liquid chromatography (HPLC) on a glycopeptide teicoplanin-based chiral stationary phase (Chirobiotic T), coupled to atomic fluorescence spectrometry (AFS) and inductively coupled plasma (ICP) MS detectors has been performed. The coupling HPLC-microwave-assisted digestion hydride generation requires on-line post-column analytes treatment, and a severe sample clean-up for fat and proteins elimination using centrifugation and ultrafiltration. Underivatized
-selenomethionine enantiomers were completely resolved in 10 min using unbuffered water mobile phase at 1 ml min−1 flow. Good selectivity and sensitivities (detection limits 3.1 and 3.5 ng ml−1 as Se for
- and
-selenomethionine, respectively) were obtained, and method robustness and simplicity, together to the low cost of AFS detector, makes it suitable for infant milk routine analysis. HPLC–ICP-MS coupling exhibits very low detection limits (0.9 ng ml−1, as Se) for each
-selenomethionine enantiomers, but the method suffers from matrix influence, that produces a poor S/N ratio and low reliability.
The methods were applied to breast and formula milk samples with recoveries of 80% of the total selenium presence, which is attributable to the existence of other unknown species. -Selenomethionine was the only isomer present in breast milk, but a 30% of -selenomethionine was also detected in formula milk. 相似文献
130.
Molecular chaperones, folding catalysts, and the recovery of active recombinant proteins fromE. coli
Jeffrey G. Thomas Amanda Ayling François Baneyx 《Applied biochemistry and biotechnology》1997,66(3):197-238
The high-level expression of recombinant gene products in the gramnegative bacteriumEscherichia coli often results in the misfolding of the protein of interest and its subsequent degradation by cellular proteases or its deposition
into biologically inactive aggregates known as inclusion bodies. It has recently become clear that in vivo protein folding
is an energy-dependent process mediated by two classes of folding modulators. Molecular chaperones, such as the DnaK-DnaJ-GrpE
and GroEL-GroES systems, suppress off-pathway aggregation reactions and facilitate proper folding through ATP-coordinated
cycles of binding and release of folding intermediates. On the other hand, folding catalysts (foldases) accelerate rate-limiting
steps along the protein folding pathway such as thecis/trans isomerization of peptidyl-prolyl bonds and the formation and reshuffling of disulfide bridges. Manipulating the cytoplasmic
folding environment by increasing the intracellular concentration of all or specific folding modulators, or by inactivating
genes encoding these proteins, holds great promise in facilitating the production and purification of heterologous proteins.
Purified folding modulators and artificial systems that mimic their mode of action have also proven useful in improving the
in vitro refolding yields of chemically denatured polypeptides. This review examines the usefulness and limitations of molecular
chaperones and folding catalysts in both in vivo and in vitro folding processes. 相似文献