共查询到20条相似文献,搜索用时 15 毫秒
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Soybean gamma-conglycinin was isolated by isoelectric precipitation and ammonium sulphate fractionation. The crude protein was purified by ion-exchange chromatography on DEAE-Sepharose CL-6B and gel filtration on Sepharose CL-6B. The purified gamma-conglycinin was homogeneous on two kinds of gel electrophoresis and an ultracentrifugal analysis. A subunit band, distinguishable from other subunit bands of beta-conglycinin and glycinin, was detected by sodium dodecyl sulphate electrophoresis. Amino acid composition was similar to those of the other storage proteins of soybean. Some physical properties were also studied. 相似文献
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A new aminopeptidase was isolated from Agave americana by fractionation, chromatography and gel filtration. The mw of the enzyme, determined by different procedures was 86000 ± 1500; the enzyme had a sedimentation coefficient of 4.96 S, a diffusion coefficient of 5.2 × 10?7 cm2/sec, a Stokes radius of 3.8 nm, a partial specific volume of 0.733 cm3/g, a frictional ratio of 1.40, a molecular absorbancy index at 280 nm of 8.36 × 104, an isoelectric point of 4.53 and contained 1.25% carbohydrate. The amino acid composition of the enzyme was determined and the aminoterminal residue was identified as lysine whilst the carboxylterminal residue was either leucine or isoleucine. No subunit structure was observed for the enzyme. 相似文献
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F W Wagner L E Ray M A Ajabnoor P E Ziemba R L Hall 《Archives of biochemistry and biophysics》1979,197(1):63-72
The extracellular aminopeptidase from Bacillus subtilis was purified 300-fold by a simple procedure which gave a high recovery of enzyme. The native enzyme was shown to be a monomer of molecular weight 46,500 and to contain 1 g-atom of Zn2+ per mole of protein. Amino acid analyses demonstrated the protein to be rich in acidic residues and Lys, to possess about 3 residues of Met, and to be devoid of Cys. When activated with 5 mm Co(NO3)2 for 90 min the activity of the native enzyme was increased; the amount of activation depended on the identity of the substrate. Cobalt activation involved the reversible binding of 1 g-atom of Co2+ per mole of protein, without displacing the native Zn2+; KCo was 1.25 mm. Zinc ions competed with Co2+ during activation, a process characterized by a KZn of 28 μm. Ions other than Co2+ did not appreciably activate the enzyme. 相似文献
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The preparation and some physical properties of sulfhemoglobin. 总被引:1,自引:0,他引:1
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Human prolactin: isolation and some properties 总被引:5,自引:0,他引:5
U J Lewis R N Singh B K Seavey 《Biochemical and biophysical research communications》1971,44(5):1169-1176
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The absorption spectrum of cobalt(II)-substituted Aeromonas aminopeptidase is markedly perturbed by the presence of equimolar concentrations of D-amino acid hydroxamates and acyl hydroxamates that have previously been shown to be powerful inhibitors of this enzyme (Wilkes, S. H., and Prescott, J. M. (1983) J. Biol. Chem. 258, 13517-13521). D-Valine hydroxamate produces the most distinctive perturbation, splitting the characteristic 527 nm absorption peak of the cobalt enzyme to form peaks at 564, 520, and 487 nm with molar extinction values of 126, 98, and 67 M-1 cm-1, respectively. A qualitatively similar perturbation, albeit with lower extinction values, results from the addition of D-leucine hydroxamate, whereas D-alanine hydroxamate perturbs the spectrum, but does not evoke the peak at 564 nm. In contrast, hydroxamates of L-valine and L-leucine in concentrations equi-molar to that of the enzyme produce only faint indications of change in the spectrum, but the hydroxamates of several other L-amino acids perturb the spectrum essentially independently of the identity of the side chain and in a qualitatively different manner from that of D-valine hydroxamate and D-leucine hydroxamate. At the high enzyme:substrate ratios used in the spectral experiments, L-leucine hydroxamate and L-valine hydroxamate proved to be rapidly hydrolyzed, hence their inability to perturb the spectrum of the cobalt-substituted enzyme during the time course of a spectral experiment. Values of kcat for L-amino acid hydroxamates, all of which are good reversible inhibitors of the hydrolysis of L-leucine-p-nitroanilide by Aeromonas aminopeptidase, were found to range from 0.01 min-1 to 5.6 min-1 for the native enzyme and from 0.27 min-1 to 108 min-1 for the cobalt-substituted enzyme; their km values toward the cobalt aminopeptidase range from 1.2 X 10(-7) M to 1.9 X 10(-5) M. The mutual exclusivity of binding for hydroxamate inhibitors and 1-butaneboronic acid, previously shown by kinetics (Baker, J. O., Wilkes, S. H., Bayliss, M. E., and Prescott, J. M. (1983) Biochemistry 22, 2098-2103), was reflected in the characteristic spectra produced by these two types of inhibitors. 相似文献
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H-lysin from Aeromonas salmonicida has been purified 1770-fold by freeze fractionation, ammonium sulphate precipitation, ion exchange chromatography and gel filtration chromatography. The purified material was predominantly H-lysin, devoid of detectable T-lysin, caseinase or gelatinase activity, although glycerophospholipid: cholesterol acyltransferase (GCAT) activity was present. The results suggested that H-lysin and GCAT activities were due to different extracellular products. Studies of the kinetics of haemolysis indicated that the H-lysin had an enzymic mode of action, and that initial erythrocyte damage appeared to precede lysis of the cell. The H-lysin was lethal to cultured rainbow trout gonad cells and leucocytes, but when it was injected intravenously in rainbow trout no pathological effects were observed. 相似文献
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Pear malic enzyme: some physical and immunochemical properties. 总被引:1,自引:0,他引:1
C J Hartmann A G Drouet 《Revue canadienne de biologie / éditée par l'Université de Montréal》1979,38(2):101-104
Some physical and immunochemical characteristics of NADP malic enzyme from climacteric pear (Pyrus communis L. var Passe-Crassane) are described. The enzyme which has a molecular weight of about 224 000 d and a sedimentation coefficient of about 9.6 S, is formed by the association of four sub-units. We were not able to isolate isozymes by electrophoresis or by immunochemistry. 相似文献
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C Kettner J Rodriguez-Absi G I Glover J M Prescott 《Archives of biochemistry and biophysics》1974,162(1):56-63
A competitive inhibitor for Aeromonas aminopeptidase has been prepared from the bromomethyl ketone derived from t-butyloxycarbonyl-l-leucine and successfully coupled to aminomethyl cellulose to form an adsorbent for affinity chromatography. The blocked form of the inhibitor was coupled to aminomethyl cellulose and then deblocked in aqueous trifluoroacetic acid to yield an insolubilized analog of an NH2-terminal l-leucyl residue. This material was effective in binding the aminopeptidase and separating it from a contaminating endopeptidase, which has a similar isoelectric point and size. Separation of the aminopeptidase and endopeptidase was shown to be due to the specificity of the affinity adsorbent for the aminopeptidase, inasmuch as separation of the enzymes did not occur on a typical anion exchange column or on a hydrophobic column lacking a free amino group. 相似文献
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A reproducible procedure for the isolation, from human placenta, of a cathepsin B1 in a homogeneous state, demonstrated by electrophoretic, ultracentrifugal and enzymic criteria, was carried out. The pH optimum was near pH5.5. The placental enzyme catalysed the release of acid-soluble u.v.-dense products from haemoglobin and myoglobin. It was inhibited by heavy metals and several compounds which react with the thiol groups. The optimum temperature was between 37° and 42°C. The molecular weight of the enzyme was calculated to be 24250. 相似文献
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Enrichment, isolation and some properties of methane-utilizing bacteria 总被引:83,自引:0,他引:83