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1.
Xylanase-induced reduction of chlorine dioxide consumption during elemental chlorine-free bleaching of different pulp types 总被引:4,自引:0,他引:4
Andreas M. Madlala Sadhvir Bissoon Suren Singh Lew Christov 《Biotechnology letters》2001,23(5):345-351
The potential of crude xylanase from Thermomyces lanuginosus and Xylanase P (a commercial xylanase) was evaluated in bleaching of various paper pulp types. Xylanases released chromophores and reducing sugars and decreased kappa number of pulps. Chlorine-bleached, alkali-extracted bagasse and post-oxygen kraft pulps, pretreated with enzymes, gained over 5 brightness points over controls. Biobleaching of soda-aq pulp with Xylanase P produced chlorine dioxide savings of up to 30% or 4.5 kg chlorine dioxide t–1 pulp. 相似文献
2.
Imines or Schiff bases formed through the interaction of reducing sugars with amino acids are known to play a critical role not only in initiating the Maillard reaction but also in its propagation through isomerization reactions initiated by the intermediate oxazolidin-5-one. FTIR spectroscopic evidence for the formation of this intermediate in a phenylalanine-glycolaldehyde model system was provided by taking advantage of a strong absorption band centered at 1778 cm−1. The identity of this peak was confirmed by observing a shift to 1736 cm−1 when [13C-1]phenylalanine was used. The intensity of this peak decreased over time with concomitant increase of two bands in the carbonyl absorption region, one centered at 1730 and the other at 1720 cm−1. The former band was shifted to 1685 cm−1, while the band at 1720 remained unchanged when [13C-1]phenylalanine was used. The simultaneous formation of a carboxylic acid and a carbonyl band is consistent with the formation of an Amadori rearrangement product. Furthermore, time-dependent analysis of the formation and decomposition of the oxazolidin-5-one intermediate suggests that it is an important precursor of the Amadori rearrangement product. In addition, through the use of appropriate model systems, [15N]phenylalanine and second-derivative spectral analysis, evidence was also provided for the formation of decarboxylated imines at 80 °C. 相似文献
3.
Flavius C. Pascut Spandan Kalra Vinoj George Nathan Welch Chris Denning Ioan Notingher 《Biochimica et Biophysica Acta (BBA)/General Subjects》2013
Background
Online label-free monitoring of in-vitro differentiation of stem cells remains a major challenge in stem cell research. In this paper we report the use of Raman micro-spectroscopy (RMS) to measure time- and spatially-resolved molecular changes in intact embryoid bodies (EBs) during in-vitro cardiogenic differentiation.Methods
EBs formed by aggregation of human embryonic stem cells (hESCs) were cultured in defined medium to induce differentiation towards cardiac phenotype and maintained in purpose-built micro-bioreactors on the Raman microscope for 5 days (between days 5 and 9 of differentiation) and spatially-resolved spectra were recorded at 24 h intervals.Results
The Raman spectra showed that the onset of spontaneous beating of EBs at day 7 coincided with an increase in the intensity of the Raman bands at 1340 cm− 1, 1083 cm− 1, 937 cm− 1, 858 cm− 1, 577 cm− 1 and 482 cm− 1. The spectral maps corresponding to these bands had a high positive correlation with the expression of the cardiac-specific α-actinin obtained by immuno-fluorescence imaging of the same EBs. The spectral markers obtained here are also in agreement with previous studies performed on individual live hESC-derived CMs.Conclusions
The intensity profile of these Raman bands can be used for label-free in-situ monitoring of EBs to estimate the efficacy of cardiogenic differentiation.General significance
As the acquisition of the time-course Raman spectra did not affect the viability or the differentiation potential of the hESCs, this study demonstrates the feasibility of using RMS for on-line non-invasive continuous monitoring of such processes inside bioreactor culture systems. 相似文献4.
Stephan BeilAnna Schamberger Wolfgang NaumannSusanne Machill Karl-Heinz van Pée 《Carbohydrate polymers》2012,87(1):117-122
A new method for the determination of the degree of N-acetylation (DA) of chitin and chitosan is described using first derivative diamond ATR FTIR spectroscopy. Applying the derivative values of the amide III band at 1327 cm−1 and the CH deformation band of the N-acetyl group at 1383 cm−1 as measure of the N-acetyl content of the sample in relation to the derivative value of the bridge oxygen vibration at 1163 cm−1 as internal standard, a linear correlation to the results of first derivative UV spectroscopy was obtained and confirmed by elemental analysis and Raman spectroscopy. The described method allows the determination of the degree of N-acetylation of chitosan and chitin in the presence of water thus making drying procedures unnecessary. 相似文献
5.
The copper(II) complex with tolfenamic acid [Cu(tolf)2(H2O)]2 was studied by X-band and K-band EPR spectroscopies in the temperature range from 90 to 300 K. The Cu2+ ions in dinuclear complex show a strong antiferromagnetic exchange interaction with |J| = 292 cm−1. The EPR spectra, which were observed for [Cu(tolf)2(H2O)]2, are typical powder spectra of the copper pairs. The spectra exhibit the hyperfine structure in low temperature range. The values of the spin-Hamiltonian parameters were determined on the basis of the best fit for the simulated spectra at both K-band (0.75 cm−1) at T = 298 K and X-band (0.3 cm−1) at T = 93 K as compared with the experimentally observed spectra. These values show that the local environment around the copper species is distorted tetragonal pyramid. This EPR evidence is consistent with the crystallographic data. 相似文献
6.
V W Yang Z Zhuang G Elegir T W Jeffries 《Journal of industrial microbiology & biotechnology》1995,15(5):434-441
ABacillus sp (V1-4) was isolated from hardwood kraft pulp. It was capable of growing in diluted kraft black liquor at pH 11.5 and produced 49 IU (mol xylose min–1 ml–1) of xylanase when cultivated in alkaline medium at pH 9. Maximal enzyme activity was obtained by cultivation in a defined alkaline medium with 2% birchwood xylan and 1% corn steep liquor at pH 9, but high enzyme production was also obtained on wheat bran. The apparent pH optimum of the enzyme varied with the pH used for cultivation and the buffer system employed for enzyme assay. With cultivation at pH 10 and assays performed in glycine buffer, maximal activity was observed at pH 8.5; with phosphate buffer, maximal activity was between pH 6 and 7. The xylanase temperature optimum (at pH 7.0) was 55°C. In the absence of substrate, at pH 9.0, the enzyme was stable at 50°C for at least 30 min. Elecrophoretic analysis of the crude preparation showed one predominant xylanase with an alkaline pl. Biobleaching studies showed that the enzyme would brighten both hardwood and softwood kraft pulp and release chromophores at pH 7 and 9. Because kraft pulps are alkaline, this enzyme could be used for prebleaching with minimal pH adjustment. 相似文献
7.
Cadena EM Du X Gellerstedt G Li J Fillat A García-Ubasart J Vidal T Colom JF 《Bioresource technology》2011,102(4):3911-3917
Flax soda/AQ pulps were treated with different fungal laccase-mediator combinations followed by physical and chemical characterization of the pulps to obtain a thorough understanding of the laccase/mediator effects on hexenuronic acid (HexA) removal and the coupling of mediator onto pulps for fiber functionalization. Large differences were found and the presence of lauryl gallate (LG) during Trametes villosa laccase (TvL) treatment (TvL + LG) resulted in a much larger reduction of pulp-linked HexA than the combination of p-coumaric acid (PCA) and Pycnoporus cinnabarinus laccase (PcL). A major portion of LG became attached to the pulp as revealed by an increase in the kappa number and further confirmed by thioacidolysis and 1H NMR analysis of solubilized pulp fractions. Additional experiments with other chemical pulps and isolated pulp xylan and lignin revealed that HexA seems to be the sole pulp component attacked by TvL + LG. As a substrate for TvL, the reaction preference order is PCA > HexA > LG. 相似文献
8.
The electrochemical and photophysical properties of two bis-nitrilo ruthenium(II) complexes formulated as [Ru(bpy)2(L)2](PF6)2, where bpy is 2,2′-bipyridine and L is AN = CH3CN and sn = NC-CH2CH2-CN, have been investigated. Electrochemical data are typical of Ru-bpy complexes with two reversible reduction peaks located near −1.3 and −1.6 V assigned to each bipyridine ligand and one RuII/RuIII oxidation wave centered at approximately +1.5 V. The sn derivative is both IR and Raman active with its coordinated CN stretch appearing at 2277 cm−1 and 2273 cm−1, respectively. The UV/Vis absorption spectrum of the sn derivative is dominated by an intense (εmax ∼ 58700 M−1 cm−1) absorption band at 287 nm assigned as a LC (π → π∗) transition. The peak observed at 418 nm (ε ∼ 10 400 M−1 cm−1) is an MLCT band while the one at 244 nm (ε ∼ 23 600 M−1 cm−1) is of LMLCT character. The AN derivative behaves similarly. Both complexes show low-temperature emission at around 537 nm with a lifetime near 10.0 μs. 1H and 13C assignments are consistent with the formulation of the complexes. The complexes undergo photosubstitution of solvent with quantum efficiencies near one. Calculated and experimental results support replacement of the nitrile ligands by solvent. Based on DFT calculations, the electron density of the HOMO lies on the metal center, the bipyridine ligands and the nitrile ligands and electron density of the LUMO resides primarily on the bipyridine ligands. The electronic spectra obtained from TDDFT calculations closely match the experimental ones. 相似文献
9.
The effects of high external pressures on the principal IR and Raman bands of crystalline dichloro(1,5-cycloctadiene)platinum(II), Pt(COD)Cl2 (COD = η4-C8H12), have been investigated for pressures up to ∼30 kbar by diamond-anvil cell microspectroscopy. This square-planar Pt(II) complex does not undergo any pressure-induced structural change throughout the pressure range investigated and the pressure dependences (dν/dP) for the vibrational modes range from −0.18 to 0.79 cm−1 kbar−1. The negative dν/dP value observed for the IR band at 1426 cm−1 (−0.18 cm−1 kbar−1) suggests that this band is chiefly associated with a CC stretching mode of the Pt-COD group. This observation provides yet another indication that high-pressure vibrational spectroscopy may indeed be a general method for establishing the presence of π-backbonding in organometallic complexes, such as metal carbonyls and alkenes. 相似文献
10.
Joshua Telser Kun-Yuan Chen Dmitry Smirnov J. Krzystek 《Journal of inorganic biochemistry》2009,103(4):487-495
Aminocarboxylate complexes of vanadium(III) are of interest as models for biologically and medicinally relevant forms of this interesting and somewhat neglected ion. The V(III) ion is paramagnetic, but not readily suited to conventional EPR, due to its integer-spin ground state (S = 1) and associated large zero-field splitting (zfs). High-frequency and -field EPR (HFEPR), however, has the ability to study such systems effectively. Three complexes, all previously structurally characterized: Na[V(trdta)] · 3H2O, Na[V(edta)(H2O)] · 3H2O, and [V(nta)(H2O)3] · 4H2O (where trdta stands for trimethylenediamine-N,N,N′,N′-tetraacetate and nta stands for nitrilotriacetate) were studied by HFEPR. All the investigated complexes produced HFEPR responses both in the solid state, and in aqueous solution, but those of [V(nta)(H2O)3] · 4H2O were poorly interpretable. Analysis of multi-frequency HFEPR spectra yielded a set of spin Hamiltonian parameters (including axial and rhombic zfs parameters: D and E, respectively) for these first two complexes as solids: Na[V(trdta)] · 3H2O: D = 5.60 cm−1, E = 0.85 cm−1, g = 1.95; Na[V(edta)(H2O)] · 3H2O: D = 1.4 cm−1, E = 0.14 cm−1, g = 1.97. Spectra in frozen solution yielded similar parameters and showed multiple species in the case of the trdta complex, which are the consequence of the flexibility of this ligand. The EPR spectra obtained in frozen aqueous solution are the first, to our knowledge, of V(III) in solution in general and show the applicability of HFEPR to these systems. In combination with very insightful previous studies of the electronic absorption of these complexes which provided ligand-field parameters, it has been possible to describe the electronic structure of V(III) in [V(trdta)]− and [V(edta)(H2O)]−; the quality of data for [V(nta)(H2O)3] does not permit analysis. Qualitatively, six-coordinate V(III) complexes with O,N donor atoms show no electronic absorption band in the NIR region, and exhibit relatively large magnitude zfs (D ? 5 cm−1), while analogous seven-coordinate complexes do have a NIR absorption band and show relatively small magnitude zfs (D < 2 cm−1). 相似文献
11.
Investigation of lignin-carbohydrate complexes in kraft pulps by selective enzymatic treatments 总被引:1,自引:0,他引:1
The occurrence of covalent bonds between residual lignin and polysaccharides in birch and pine kraft pulps was investigated
by specific enzymatic treatments. Pure enzymes degrading cellulose, xylan and mannan were used both separately and in combination.
Comparison of the molar masses of polysaccharides and lignin in the orginal pulps and in the residual pulps after enzymatic
treatments showed that residual lignin in birch kraft pulp is linked at least to xylan. A minor portion may also be linked
to cellulose. In pine kraft pulp some of the residual lignin appears to be linked to cellulose, glucomannan and xylan. The
linkages between lignin and cellulose and hemicelluloses may be either native or formed during pulp processing. The results
also provided new information on the synergistic action of cellulose- and hemicellulose-degrading enzymes on pulp fibres.
The synergism appears to be mainly due to the structure of the pulp fibres, with different layers of cellulose sheets, hemicelluloses
and lignin. On the other hand the results also provided information about fibre structure. The degradation of xylan clearly
enhanced the action of enzymes on cellulose, suggesting that xylan partially covers the cellulose. A similar phenomenon was
not observed in the simultaneous hydrolysis of glucomannan and cellulose. However, the results suggest that glucomannan does
interact with cellulose, possibly by non-covalent linkages.
Received: 8 July 1998 / Received revision: 7 October 1998 / Accepted: 11 October 1998 相似文献
12.
A. Flores-MoralesR. Mora-Escobedo 《Carbohydrate polymers》2012,87(1):61-68
The nixtamalization, production and storage of tortillas in refrigeration cause several changes on the starch structure, resulting in an increased crystallinity and therefore a higher content of resistant starch. The IR analysis for resistant starch (RS) showed a band at 1047 cm−1 associated to the retrogradation process; this band was due to the weakening of the intermolecular H-bonds. These associated together to form ordered regions. The Raman analysis shows a characteristic band at 856 cm−1 corresponding to C-C skeletal modes of glucose of α-1,4 glycosidic linkage starches, and a band at 480 cm−1 attributed to skeletal vibrations of the pyranose ring in the glucose unit of starches. These changes may be related to the polymerization degree of the starch molecules, as well as to the retrogradation of amylose and amylopectin. The spectrum of 13C CP-MAS/NMR for RS3 supports the results obtained by IR and Raman. Lipidic and proteic groups were observed which may be in the form of complexes with amylose. One can proclaim that the existence of the salt form is induced and stabilized by the interactions dominating the V amylose structure in the solid state. 相似文献
13.
Fluorescence line-narrowing (FLN) spectroscopy at 10 K was used to study the effect of high pressure through the prosthetic group in horseradish peroxidase (HRP), which was Mg-mesoporphyrin (MgMP) replacing the heme of the enzyme. The same measurement was performed on MgMP in a solid-state amorphous organic matrix, dimethyl sulfoxide (DMSO). Series of FLN spectra were registered to determine the (0, 0) band shape through the inhomogeneous distribution function (IDF). In the range of 0-2 GPa a red-shift of the IDF was determined, and yielded the isothermal compressibility of MgMP-HRP as 0.066 GPa−1, which is significantly smaller than that found earlier as 0.106 GPa−1 by fine-tuning the pressure in the range up to 1.1 MPa. The vibrational frequencies also shifted with pressure increase, as expected. The compressibility in the DMSO matrix was smaller, 0.042 GPa−1, both when the pressure was applied at room temperature before cooling to 10 K, or at 10 K. At 200 K or above, the bimodal (0, 0) band shape in DMSO showed a population conversion under pressure that was not observed at or below 150 K. A significant atomic rearrangement was estimated from the volume change, 3.3 ± 0.7 cm3/mol upon conversion. The compressibility in proteins and in amorphous solids seems not to significantly depend on the temperature and in the protein it decreases toward higher pressures. 相似文献
14.
Hiroshi Sakiyama Tetsuya Suzuki Rie Ito Mikio Yamasaki 《Inorganica chimica acta》2005,358(6):1897-1903
Dinuclear nickel(II) complexes [Ni2(bomp)(MeCO2)2]BPh4 (1) and [Ni2(bomp)(PhCO2)2]BPh4 (2) were synthesized with the dinucleating ligand 2,6-bis[bis(2-methoxyethyl)aminomethyl]-4-methylphenol [H(bomp)]. X-Ray analysis revealed that the complex 1 · 0.5CHCl3 contains two nickel(II) ions bridged by phenolic oxygen and two acetate groups, forming a μ-phenoxo-bis(μ-acetato)dinickel(II) core. Electronic spectra were investigated for 1 and 2 in the range of 400-1800 nm, and the data were typical for the octahedral high-spin nickel(II) complexes. Obtained spectral components were well simulated based on the angular overlap model assuming the trigonally distorted octahedral geometry. Magnetic susceptibility was measured for 1 and 2 over a temperature range of 4.5-300 K. The optimized magnetic data were J = 1.75 cm−1, zJ′ = −0.234 cm−1, g = 2.21, D = 15.1 cm−1, and TIP = 370 × 10−6 cm−1 for complex 1 and J = 3.55 cm−1, zJ′ = −0.238 cm−1, g = 2.23, D = 21.8 cm−1, and TIP = 470 × 10−6 cm−1 for complex 2. The data revealed ferromagnetic interactions between the two nickel(II) ions. 相似文献
15.
A novel assay method was investigated for urease (EC 3.5.1.5) from Pseudomonas aeruginosa and Canavalia ensiformis by Fourier transform infrared spectroscopy. This enzyme catalyzed the hydrolysis of urea in phosphate buffer in deuterium oxide (2H2O). The intensities of the bicarbonate bands maxima at 1625 and 1365 cm−1 and of the amide I band at 1605 cm−1 were measured as a function of time to study the kinetics of urea hydrolysis. The extinction coefficients ε of urea and bicarbonate were determined to be 0.72, 0.48, and 0.56 mM−1 cm−1 at 1625, 1605, and 1365 cm−1, respectively. The initial velocity is proportional to the enzyme concentration by using the ureases from both C.ensiformis and P. aeruginosa. The kinetic constants (Vmax, Km, and Kcat) determined by Lineweaver-Burk plot were 532.2 U mg−1 protein, 6.4 mM, and 806.36 s−1, respectively. These data are in agreement with the results obtained by a spectrophotometric method using a linked assay based on glutamate dehydrogenase in aqueous media. Therefore, this spectroscopic method is highly suited to assay for urease activity and its kinetic parameters by using either cell-free extracts or purified enzyme preparations with an additional advantage of performing a real-time measurement of urease activity. 相似文献
16.
Rupenyan A van Stokkum IH Arents JC van Grondelle R Hellingwerf K Groot ML 《Biophysical journal》2008,94(10):4020-4030
Proteorhodopsin is an ion-translocating member of the microbial rhodopsin family. Light absorption by its retinal chromophore initiates a photocycle, driven by trans/cis isomerization, leading to transmembrane translocation of a proton toward the extracellular side of the cytoplasmic membrane. Here we report a study on the photoisomerization dynamics of the retinal chromophore of proteorhodopsin, using femtosecond time-resolved spectroscopy, by probing in the visible- and in the midinfrared spectral regions. Experiments were performed both at pH 9.5 (a physiologically relevant pH value in which the primary proton acceptor of the protonated Schiff base, Asp97, is deprotonated) and at pH 6.5 (with Asp97 protonated). Simultaneous analysis of the data sets recorded in the two spectral regions and at both pH values reveals a multiexponential excited state decay, with time constants of ∼0.2 ps, ∼2 ps, and ∼20 ps. From the difference spectra associated with these dynamics, we conclude that there are two chromophore-isomerizaton pathways that lead to the K-state: one with an effective rate of ∼(2 ps)−1 and the other with a rate of ∼(20 ps)−1. At high pH, both pathways are equally effective, with an estimated quantum yield for K-formation of ∼0.7. At pH 6.5, the slower pathway is less productive, which results in an isomerization quantum yield of 0.5. We further observe an ultrafast response of residue Asp227, which forms part of the counterion complex, corresponding to a strengthening of its hydrogen bond with the Schiff base on K-state formation; and a feature that develops on the 0.2 ps and 2 ps timescale and probably reflects a response of an amide II band in reaction to the isomerization process. 相似文献
17.
Investigations of vibrational coherence in the low-frequency region of ferric heme proteins
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Femtosecond coherence spectroscopy is applied to a series of ferric heme protein samples. The low-frequency vibrational spectra that are revealed show dominant oscillations near 40 cm−1. MbCN is taken as a typical example of a histidine-ligated, six-coordinate, ferric heme and a comprehensive spectroscopic analysis is carried out. The results of this analysis reveal a new heme photoproduct species, absorbing near 418 nm, which is consistent with the photolysis of the His93 axial ligand. The photoproduct undergoes subsequent rebinding/recovery with a time constant of ∼4 ps. The photoproduct lineshapes are consistent with a photolysis quantum yield of 75-100%, although the observation of a relatively strong six-coordinate heme coherence near 252 cm−1 (assigned to ν9 in the MbCN Raman spectrum) suggests that the 75% lower limit is much more likely. The phase and amplitude excitation profiles of the low-frequency mode at 40 cm−1 suggest that this mode is strongly coupled to the MbCN photoproduct species and it is assigned to the doming mode of the transient penta-coordinated material. The absolute phase of the 40 cm−1 mode is found to be π/2 on the red side of 418 nm and it jumps to 3π/2 as excitation is tuned to the blue side of 418 nm. The absolute phase of the 40 cm−1 signal is not explained by the standard theory for resonant impulsive stimulated Raman scattering. New mechanisms that give a dominant momentum impulse to the resonant wavepacket, rather than a coordinate displacement, are discussed. The possibilities of heme iron atom recoil after photolysis, as well as ultrafast nonradiative decay, are explored as potential ways to generate the strong momentum impulse needed to understand the phase properties of the 40 cm−1 mode. 相似文献
18.
Priti KatroliaPeng Zhou Pan ZhangQiaojuan Yan Yinan LiZhengqiang Jiang Haibo Xu 《Carbohydrate polymers》2012,87(1):480-490
A novel β-mannanase gene (CsMan5A) was cloned from Chaetomium sp. CQ31 and expressed in Pichia pastoris. It had an open reading frame of 1251 bp encoding 416 amino acids and contained two introns. The deduced amino acid sequence shared the highest similarity (73%) with the β-mannanase from Emericella nidulans and belongs to glycosyl hydrolase family 5. The recombinant β-mannanase (CsMan5A) was secreted at extremely high levels of 50,030 U mL−1 and 6.1 mg mL−1 in high cell density fermentor. The purified enzyme was optimally active at pH 5.0 and 65 °C and displayed broad pH stability (pH 5.0-11.0) and exhibited specificity towards locust bean gum (Km = 3.1 mg mL−1), guar gum (Km = 9.3 mg mL−1) and konjac powder (Km = 10.5 mg mL−1). It efficiently degraded mannan polysaccharides into mannose and mannooligosacccharides, and also hydrolyzed mannotriose and mannotetraose. These properties make CsMan5A highly useful in food, feed and paper/pulp industries. 相似文献
19.
Cristian Enachescu Itana Krivokapic Jose Antonio Real Andreas Hauser 《Inorganica chimica acta》2007,360(13):3945-3950
The spin transition of the [Co(terpy)2]2+ complex (terpy = 2,2′:6′,2″-terpyridine) is analysed based on experimental data from optical spectroscopy and magnetic susceptibility measurements. The single crystal absorption spectrum of [Co(terpy)2](ClO4)2 shows an asymmetric absorption band at 14 400 cm−1 with an intensity typical for a spin-allowed d-d transition and a temperature behaviour typical for a thermal spin transition. The single crystal absorption spectra of suggest that in this compound, the complex is essentially in the high-spin state at all temperatures. However, the increase in intensity observed in the region of the low-spin MLCT transition with increasing temperature implies an unusual partial thermal population of the low-spin state of up to about 10% at room temperature. Finally, high-spin → low-spin relaxation curves following pulsed laser excitation for [Co(terpy)2](ClO4)2 dispersed in KBr discs, and as a comparison for the closely related [Co(4-terpyridone)2](ClO4)2 spin-crossover compound are given. 相似文献
20.
Piotr K. Grzyska Robert P. Hausinger Denis A. Proshlyakov 《Analytical biochemistry》2010,399(1):64-548
The addition of divalent metal ions or substrate taurine to TauD, an α-ketoglutarate-dependent dioxygenase, alters its UV absorption, as clearly observed by monitoring the protein’s difference spectra. Binding of metal ions leads to a decrease in absorption at ∼297 nm and modulation of other features. A separate signature with enhanced absorption at ∼295 nm is identified for binding of taurine. These narrow (∼700 cm−1) and intense (∼0.5 mM−1 cm−1) spectral changes are attributed to ligand-induced protein conformational changes affecting the environment of aromatic residues. The changes in the UV difference spectra were exploited to assess directly the thermodynamics and kinetics of ligand interactions in wild-type TauD and selected variants. This approach holds promise as a new tool to probe ligand-induced conformational changes in a wide range of other proteins. Experimental and quantification approaches for a reliable analysis of protein absorption below 320 nm are presented. 相似文献