puri † jörg rocholl ‡ and sascha steffen§

Báo cáo khoa học: Human recombinant prolidase from eukaryotic and prokaryotic sources Expression, puriﬁcation, characterization and long-term stability studies pptx

Ngày tải lên : 07/03/2014, 11:20

... duplication and of a mutation generating symptomatic and asymptomatic outcome within the same family J Med Genet, in press Fernandez-Espla MD, Martin-Hernandez MC & Fox PF (1997) Puri cation and characterization ... endogenous enzyme and the tagged and untagged recombinant prolidase was close to the activity measured on day (123%, 90% and 85%, respectively) This paper describes the puri cation and characterization ... similarity of the recombinant and endogenous prolidase in terms of substrate specificity, optimal pH and temperature for activity, and metal dependence is discussed, and a detailed analysis of long-term...

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Báo cáo khoa học: Puriﬁcation, microsequencing and cloning of spinach ATP-dependent phosphofructokinase link sequence and function for the plant enzyme pdf

Ngày tải lên : 07/03/2014, 11:20

... yielded 309-fold puri ed ATP-PFK, but was accompanied by the loss of 93% of total activity (Table 1) Results and Discussion Puri cation and microsequencing of spinach PFK The present puri cation protocol ... determined from the puri ed protein (Fig 2) New degenerate primers were designed against the peptides LSGNAVLGDIGVHFK and EIYFEPTK, and produced a PCR fragment of 750 bp that was cloned and sequenced ... Gly105 and Lys124, in the Esc coli enzyme [45] are conserved in the atypical and previously uncharacterized ATP-dependent PFK sequences [3,4,6,39], and they are also conserved in SoPFK1 and SoPFK2...

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Báo cáo khoa học: Puriﬁcation, characterization and molecular cloning of tyrosinase from the cephalopod mollusk, Illex argentinus docx

Ngày tải lên : 17/03/2014, 10:20

... and tyrosinases [7,9]; highly conserved amino acid sequences and residues around these copper ligands in molluscan hemocyanins and tyrosinases were also well conserved in ST94 On the other hand, ... literature [43] The Km and Vmax values for different substrates were obtained from the Hanes–Woolf equation Ó FEBS 2003 Extraction of RNA and ﬁrst strand cDNA preparation All DNA and RNA manipulations ... with sense and antisense primers corresponding to a portion of the N-terminal amino acid sequence of ST94 (MVDVSQSD) and that of ST94t (MSPQEYIQ), respectively Sense (TYF1 and TYF2) and antisense...

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Báo cáo Y học: Puriﬁcation, characterization and subunits identiﬁcation of the diol dehydratase of Lactobacillus collinoides pot

Ngày tải lên : 23/03/2014, 21:20

... four main bands with molecular masses of approximately 63, 59, 28 and 22 kDa (proteins 1, 2, and 4, respectively) (Fig 2) The molecular masses of three of these bands (proteins 1, and 4) were ... out under nondenaturing conditions and revealed one main band and two weaker bands Their dissociation by SDS in the second dimension showed that the main band was released into three subunits ... lanes 4,5,6), pQE30HisDD (lanes 7–9), total expression (lanes and 7), insoluble fraction (lanes and 8) and soluble fraction (lanes and 9) no activity was detected in the extracts even using the...

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Báo cáo khoa học: Brassinin oxidase, a fungal detoxifying enzyme to overcome a plant defense – puriﬁcation, characterization and inhibition potx

Ngày tải lên : 30/03/2014, 04:20

... Sigma prepared reagent and BSA as the standard Synthesis and spectroscopic characterization of phytoalexins and analogue library Compounds and phytoalexins shown in Tables and supplementary Table ... ‘blackleg’ fungi L maculans (L m.) and L biglobosa (L b.) Table Enzyme yields and puri cation factors for BO Recoveries are expressed as a percentage of initial activity and puri cation factors are calculated ... understand the role of BO and test potential inhibitors, the enzyme was puri ed, characterized and shown to be a novel enzyme, consistent with the unusual transformation it catalyzes (Fig 1) Puri ed...

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Báo cáo khoa học: Puriﬁcation, characterization and biosynthesis of parabutoxin 3, a component of Parabuthus transvaalicus venom pptx

Ngày tải lên : 31/03/2014, 09:20

... to different ion channels and receptors could be performed and a structural model for this novel toxin has been proposed MATERIALS AND METHODS Venom collection and puri cation P transvaalicus ... Kv1.2 and Kv1.3 peak currents by 54% and 53% at )30 mV (n ¼ 3), and by 53% and 54% (n ¼ 3) at 20 mV In the presence of 70 lM rPBTx3, the Kv1.1 peak current was blocked by 45% (n ¼ 3) at )30 mV and ... Kv1.2 and Kv1.3, V1/2 is not shifted by rPBTx3 as illustrated by the dashed lines Slope values (s) for the control and the toxin curves are, respectively, 7.5 and 9.6 for Kv1.2, and 9.3 and 8.5...

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Tài liệu Báo cáo khoa học: A facile method for expression and puriﬁcation of the Alzheimer’s disease-associated amyloid b-peptide pdf

Ngày tải lên : 18/02/2014, 13:20

... wild-type (Fig 2E) Puri cation of Ab(M1–40) and Ab(M1–42) The present work describes a rapid and inexpensive puri cation scheme to produce high-purity Ab(M1–40) and Ab(M1–42) in 24 h The puri cation ... supplied with the enzyme, and contained Aba, Abb and Abc at 40 nm each, and the start and stop primers Abstart and Abstop at 600 nm each, and 200 lm each of dATP, dCTP, dGTP and dTTP The product was ... (2% gel), and the cleaved product was puri ed using the GFX PCR and gel band puri cation kit The puri ed cut gene was ligated into PetSac vector (a modiﬁed from of Pet3a with NdeI and SacI cloning...

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Tài liệu Báo cáo khoa học: Puriﬁcation and structural characterization of a D-amino acid-containing conopeptide, conomarphin, from Conus marmoreus docx

Ngày tải lên : 18/02/2014, 17:20

... kinetics of the Trp14 and Thr15 amide protons The two observed small 3JHN-Ha coupling constants for residues Asn11 and Ser12, and the dNN(i,i + 2), daN(i,i + 2) [Phe13(CaH)–Thr15(NH)] and daN(i,i + 3) ... small 3JHN-Ha coupling constants for residues Asn11 and Ser12 and the dNN(i,i + 2), daN(i,i + 2) [Ser12(CaH)–Trp14(NH) and Asn11(CaH)–Phe13(NH)] and daN(i,i + 3) [Asn11 (CaH)–Trp14(NH)] NOEs in ... sequencing Puri cation The puri cation procedure and conditions were exactly the same as previously described [6] Brieﬂy, the crude venom was ﬁrst separated on a Sephadex G-25 column and the second...

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Tài liệu Báo cáo khoa học: Puriﬁcation and characterization of glutamate N-acetyltransferase involved in citrulline accumulation in wild watermelon doc

Ngày tải lên : 20/02/2014, 03:20

... and centrifuged Negligible loss of enzymatic activity and 24-fold puri cation were achieved Heat treatment was followed by six chromatography steps comprising hydrophobic interaction, anion and ... in wild watermelon plants Images and represent photographs of wild watermelon plants watered daily and subjected to drought for days, respectively, and images and show their thermal distributions ... Takahara et al had been strongly induced This paper reports the puri cation and characterization of GAT from wild watermelon leaves, and discusses its function during drought ⁄ strong-light stress...

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Tài liệu Báo cáo khoa học: The tungsten-containing formate dehydrogenase from Methylobacterium extorquens AM1: Puriﬁcation and properties docx

Ngày tải lên : 20/02/2014, 23:20

... describe, for the first time, puri cation and properties of a tungsten-containing NAD-dependent FDH from the aerobic bacterium, M extorquens AM1 Materials and methods Organism and growth conditions M ... Haloferax volcanii and Methanothermobacter thermautotrophicus, and a function was suggested for these genes in the specific uptake of Fig Emission fluorescence spectra of puri ed FDH1 and milk xanthine ... (both under oxic and anoxic conditions, see below) Protein was determined by the Bradford assay using Bio-Rad reagent with BSA as a standard [26] NAD-dependent FDH (FDH1) was puri ed from M extorquens...

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Tài liệu Báo cáo khoa học: Puriﬁcation and cDNA cloning of a cellulase from abalone Haliotis discus hannai ppt

Ngày tải lên : 20/02/2014, 23:20

... lyophilized and subjected to gel-filtration through Sephacryl S-200 HR Consequently, the HdEG66 was eluted in a single major peak and showed a single band of 66 000 Da in both the SDS/PAGE and zymogram ... indicates that HdEG66 is a monomeric enzyme The yield and purity of the HdEG66 in respective puri cation steps are summarized in Table The puri ed HdEG66 showed a specific activity of 13.9 UÆmg)1, ... in Table 2, and Hd5RACE-DNA and Hd3RACE-DNA for 5¢- and 3¢-terminal regions were amplified, respectively By combining the nucleotide sequences of the Hd5RACE-DNA, Hd1-DNA, Hd2DNA and the Hd3RACE-DNA...

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Tài liệu Báo cáo khoa học: Puriﬁcation and characterization of a sialic acid speciﬁc lectin from the hemolymph of the freshwater crab Paratelphusa jacquemontii pdf

Ngày tải lên : 21/02/2014, 00:20

... [30] and [63] lectin Horse and rabbit erythrocytes, which have a high content of 4-O-Ac-NeuAc and 9-O-Ac-NeuAc, respectively, showed the highest titers with the lectin Human A and O, sheep and ... 2-mercaptoethanol and 5% (v/v) glycerol] Gels were ﬁxed and stained with a solution containing 0.05% (w/v) Coomassie Blue R-250, 10% (v/v) acetic acid, and 25% (v/v) isopropyl alcohol, and destained ... CaCl2, at °C for 18 h, and then for h with fresh mM CaCl2 The dialysate was then aliquoted, lyophilized and stored at )20 °C The elution proﬁle (Fig 1) and a summary of puri cation (Table 1) give...

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Tài liệu Báo cáo Y học: A novel meta-cleavage dioxygenase that cleaves a carboxyl-groupsubstituted 2-aminophenol Puriﬁcation and characterization of 4-amino-3-hydroxybenzoate 2,3-dioxygenase from Bordetella sp. strain 10d doc

Ngày tải lên : 21/02/2014, 01:21

... 1,2,4-trihydroxybenzene and pyrogallol) in aqueous solution were always freshly prepared and used immediately Effect of various compounds on the enzyme activity The effect of metal salts, and chelating and sulfhydryl ... nicotinamide was required for growth and potassium tellurite inhibited growth Thus, strain 10d was identiﬁed as a species of Bordetella Puri cation and properties of the puri ed enzyme Fig Growth of ... at 294 nm The enzyme was puri ed 110-fold with an overall yield of 3% (Table 1) After electrophoresis, the puri ed enzyme exhibited a single protein band on both native and denaturing polyacrylamide...

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Tài liệu Báo cáo khoa học: Puriﬁcation and characterization of a membrane-bound enzyme complex from the sulfate-reducing archaeon Archaeoglobus fulgidus related to heterodisulﬁde reductase from methanogenic archaea pdf

Ngày tải lên : 21/02/2014, 03:20

... the a band at 557 nm absorption maxima at 420 nm (c band), 530 nm (b band) and 557 nm (a band) are characteristic of cytochrome b Heme was extracted from the protein with acidic acetone and the ... detected with g values at 2.031, 1.994, and 1.951 The resonance started to develop at potentials ‡ mV and was stable at potentials up to +350 mV The loss and formation of the resonance was associated ... hydrogenase and their function in fumarate and polysulﬁde respiration Arch Microbiol 170, 50–58 46 Keltjens, J.T & Vogels, G.D (1993) Conversion of methanol and methylamines to methane and carbon...

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Tài liệu Báo cáo Y học: Puriﬁcation, characterization, cloning, and expression of the chicken liver ecto-ATP-diphosphohydrolase pot

Ngày tải lên : 22/02/2014, 04:20

... Ecto-Atpases and Related Ectonucleotidases (Vanduﬀel, L & Lemmens, R., eds), pp 1–8 Shaker Publishing B.V., Maastricht, the Netherlands Handa, M & Guidotti, G (1996) Puri cation and cloning of ... pooled and desalted on Sephadex G-25 column (1.5 · 48 cm) The desalted and partially puri ed chicken liver ectoATPDase fraction ( 20 mL) was added to mL of MC22-hydrazide resin [7], and equilibrated ... primers and thermal cycling conditions but with Advantage Taq DNA polymerase (Clontech) for subsequent TA cloning The 1.5-kb PCR product was gel puri ed and ligated to pCR2.1 (Invitrogen) and an...

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Tài liệu Báo cáo Y học: Puriﬁcation, characterization, immunolocalization and structural analysis of the abundant cytoplasmic b-amylase from Calystegia sepium (hedge bindweed) rhizomes ppt

Ngày tải lên : 22/02/2014, 07:20

... Extraction and puri cation of b-amylase from rhizomes of C sepium The b-amylase was puri ed by classical protein puri cation techniques Fresh rhizomes (100 g) were cut into small pieces and homogenized ... no 210) and isoelectric focusing standards (pI 3.5–9.3) were used Total neutral sugar content of the puri ed protein was determined by the phenol/H2SO4 method [18], with D -glucose as standard ... Ireland) To determine the pH stability of the b-amylase, aliquots of a solution of the puri ed protein (4.06 mg·mL21 in water) were adjusted to different pH values in a range between and 12, and...

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Báo cáo khoa học: Puriﬁcation and characterization of zebraﬁsh hatching enzyme – an evolutionary aspect of the mechanism of egg envelope digestion pot

Ngày tải lên : 07/03/2014, 04:20

... min, bands with molecular masses of 43 and 39 kDa were observed in addition to undigested bands of ZP2 and ZP3 After incubation for 10 min, three major bands with molecular masses of 43, 39 and ... 2), 10 (lane 3) and 40 (lane 4) Numbers on the right show the molecular masses of the major bands 5938 39 kDa band disappeared, and only two bands with the same mobility as the 43 and 36.5 kDa products ... ZHE1, MHCE or MLCE, and their SDS ⁄ PAGE patterns were compared SDS ⁄ PAGE of the MHCE digest after a 10 and 40 incubation gave two bands (43 and 39 kDa), and an additional band (36.5 kDa) was...

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Báo cáo khoa học: Puriﬁcation and characterization of the cysteine proteinases in the latex of Vasconcellea spp. ppt

Ngày tải lên : 07/03/2014, 11:20

... Science and Technology in Flanders (IWT-Vlaanderen)’ and by FWO-Vlaanderen (project number 3G005100) We thank W Peumans for his extensive assistance during the protein puri cation I Vandenberghe ... CC-III and CC23 These results conﬁrm the hypothesis of Walraevens et al [17] that CC-III is a chymopapain homologue and predict VXH-C and VS-B to be the corresponding genes in V · heilbornii and ... divided into pools A and B Whereas the later fractions (pool B) revealed two protein bands of 27 and 30 kDa after SDS ⁄ PAGE, the earlier fractions (pool A) showed an extra protein band of higher molecular...

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Báo cáo khoa học: Puriﬁcation and structural analysis of the novel glycoprotein allergen Cyn d 24, a pathogenesis-related protein PR-1, from Bermuda grass pollen pot

Ngày tải lên : 07/03/2014, 12:20

... allergic to BGP require individual diagnosis and therapy, and an understanding of the structure of these allergens is essential to the improvement of diagnosis and the design of adequate therapeutic ... combination of CM-TSK and RP-HPLC The ﬁnal material was homogenous, as shown by the presence of a single band on SDS ⁄ PAGE, a single sharp peak on RP-HPLC, and a single band on immunoblotting ... Chow et al isoforms and these have high pI values ranging from 9.5 to 10.5 [10–15] Similarly, Cyn d is a calcium-binding protein containing two Ca2+, and its allergenicity and cross-reactivity...

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Báo cáo khóa học: Localization, puriﬁcation and properties of a tetrathionate hydrolase from Acidithiobacillus caldus pot

Ngày tải lên : 07/03/2014, 14:20

... Filter YM-3, and resolved by SDS/PAGE One major band with a molecular mass of 54 kDa and a few faint bands with molecular mass < 40 kDa were also detected To remove the low molecular mass impurities, ... of this work was to purify and characterize tetrathionate hydrolase from A caldus The products of the enzymatic reaction were also determined Materials and methods Bacteria and growth conditions ... demonstrated a 17-fold puri cation and about 8% recovery of the enzyme activity (Table 2) This active enzyme preparation showed only one band when stained by GelCode Blue Stain Reagent and SilverXpress...

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