scholarly journals Molecular weight of Escherichia coli β-galactosidase in concentrated solutions of guanidine hydrochloride

1970 ◽  
Vol 120 (2) ◽  
pp. 255-261 ◽  
Author(s):  
Robert P. Erickson

The molecular weight of Escherichia coli β-galactosidase was determined in 6m- and 8m-guanidine hydrochloride by meniscus-depletion sedimentation equilibrium, sedimentation velocity and viscosity. Sedimentation equilibrium revealed heterogeneity with the smallest component having a molecular weight of about 50000. At lower speeds, the apparent weight-average molecular weight is about 80000. By use of a calculation based on an empirical correlation for proteins that are random coils in 6m-guanidine hydrochloride, sedimentation velocity gave a molecular weight of 91000, and the intrinsic viscosity indicated a viscosity-average molecular weight of 84000. Heating in 6m-guanidine hydrochloride lowered the viscosity of β-galactosidase in a variable manner.

1969 ◽  
Vol 115 (4) ◽  
pp. 639-643 ◽  
Author(s):  
R. H. Villet ◽  
K. Dalziel

A method is described for the isolation of 6-phosphogluconate dehydrogenase from sheep liver. The product appears to be homogeneous in polyacrylamide-gel electrophoresis and in sedimentation-velocity and sedimentation-equilibrium studies in the ultracentrifuge. The molecular weight is estimated as 129000 from equilibrium sedimentation.


1977 ◽  
Vol 163 (3) ◽  
pp. 427-432 ◽  
Author(s):  
R H Swisher ◽  
M L Landt ◽  
F J Reithel

The weight-average molecular weight of the Mo-Fe protein isolated from Azotobacter vinelandii has been determined by sedimentation-equilibrium techniques. In buffer, the value is 245000+/-5000; in 8M-urea, the value is 61000+/-1000. The protein was separated into two components by chromatography on CM-cellulose in 7M-urea, pH 4.5. These components have similar molecular weights but were shown to differ in charge, amino acid content and arginine-containing peptides. It is proposed that the tetramer has the subunit composition (nalpha2nbeta2).


1956 ◽  
Vol 34 (4) ◽  
pp. 445-450 ◽  
Author(s):  
W. Donald Graham ◽  
Odette Patry ◽  
E. Helen Jackman

Failure to consider the presence of up to 16% by volume of methanol in solutions of dextran fractions had a very marked effect on apparent intrinsic viscosity determinations (the term apparent signifies that measurements were made assuming that the solvent was water only). Unless methanol were removed or otherwise taken into account, high erroneous results were obtained. Apparent weight average molecular weights determined by light scattering were not significantly affected at these alcohol concentrations. The relations found over the range 0 to 16% methanol for dextran samples with weight average molecular weights of 265,000, 155,000, and 72,000 held for the latter sample up to 44% methanol. In the higher range of alcohol concentration the apparent weight average molecular weight was depressed. The true intrinsic viscosity of dextran solutions decreased as methanol concentration was increased.


1973 ◽  
Vol 135 (1) ◽  
pp. 93-99 ◽  
Author(s):  
Barry J. Kitchen ◽  
Colin J. Masters ◽  
Donald J. Winzor

A purified arylesterase preparation from bovine plasma was characterized to the extent that it has a partial specific volume of 0.91ml/g and an apparent z-average molecular weight of 440000. The relatively large magnitude of the former reflects the presence of phospholipids, cholesterol, triglycerides and β-carotene, the last-named being responsible for the pronounced yellow colour of the preparation. Removal of the lipid material is accompanied by a decrease in the apparent z-average molecular weight to 120000, the size of the smallest species detected by high-speed sedimentation equilibrium being in the vicinity of 70000 daltons: denaturation of the lipid-free preparation with 6m-guanidine hydrochloride caused essentially complete breakdown into subunits of this size. In kinetic studies on the enzyme the maximal velocity for the hydrolysis of phenyl acetate was found to increase by 60% on addition of 1 mm-Ca2+, with the Km showing a concomitant decrease from 6.6 to 2.1 mm. Removal of lipid had no detectable effect on Vmax. or Km in either the presence or the absence of Ca2+. It is concluded that the bovine plasma arylesterase preparation is either a lipoprotein or an enzyme–lipoprotein complex with properties very similar to those of the α1-lipoprotein or high-density lipoprotein (HDL2) fraction of serum.


1976 ◽  
Vol 29 (6) ◽  
pp. 481
Author(s):  
EF Woods

Fractionated samples of the soluble S-carboxymethyl proteins from kookaburra beak (Frenkel and Gillespie 1976) were examined by equilibrium sedimentation. The molecular weight was found to be 11 300 when the photoelectric scanning absorption optical system was employed and 13 700 when Rayleigh interference optics were used. Possible explanations for this difference are considered and it is concluded that it must arise from heterogeneity of the protein. Optical rotatory dispersion measurements indicate that the proteins probably exist as random coils in dilute aqueous buffer.


1995 ◽  
Vol 60 (3) ◽  
pp. 489-497 ◽  
Author(s):  
Hynek Balcar ◽  
Jan Sedláček ◽  
Marta Pacovská ◽  
Vratislav Blechta

Catalytic activity of the tungsten aryloxo complexes WCl5(OAr) and WOCl3(OAr), where Ar = 4-t-C4H9C6H4, 2,6-(t-C4H9)2C6H3, 2,6-Cl2C6H3, 2,4,6-Cl3C6H2, and 2,4,6-Br3C6H2 in polymerization of phenylacetylene (20 °C, monomer to catalyst molar ratio = 1 000) was studied. The activity of WCl5(OAr) as unicomponent catalysts increases with increasing electron withdrawing character of the -OAr ligand. Addition of two equivalents of organotin cocatalysts (Me4Sn, Bu4Sn, Ph4Sn, Bu3SnH) to WCl5(O-C6H2Cl3-2,4 ,6) has only slight positive effect (slightly higher polymer yield and/or molecular weight of poly(phenylacetylene)s was achieved). However, in the case of WOCl3(O-C6H3Cl2-2, 6) catalyst, it enhances the activity considerably by eliminating the induction period. Poly(phenylacetylene)s prepared with the catalysts studied have weight-average molecular weight ranging from 100 000 to 200 000. They are trans-prevailing and have relatively low molar fraction of monomer units comprised in cyclohexadiene sequences (about 6%).


1942 ◽  
Vol 20c (3) ◽  
pp. 130-159 ◽  
Author(s):  
A. G. McCalla ◽  
Nils Gralén

The molecular characteristics of gluten in sodium salicylate solutions were studied by means of sedimentation velocity, sedimentation equilibrium, and diffusion measurements. The proportion of total gluten protein molecularly dispersed increased with increase in concentration of sodium salicylate up to 12%, but the dispersed portions had essentially the same sedimentation constant (2.5 ± 0.15) regardless of the concentration of the dispersing medium.The most soluble 25 per cent of the gluten was all molecularly dispersed, but was definitely inhomogeneous. The weight-average molecular weight of this fraction was 44,000, but there is reason to believe the minimum weight may be about 35,000. None of the other fractions was entirely molecularly dispersed, the proportion decreasing with decreasing solubility of the fractions. Aggregates of many sizes existed in all of these fractions, but only the most insoluble contained aggregates large enough to cause opacity. Sedimentation constants of the molecularly dispersed portions increased slightly with decreasing solubility, while diffusion constants decreased markedly. None of the fractions yielded normal curves (diffusion diagrams) but the more soluble the fraction, the more nearly normal the curve. The inhomogeneity responsible for the varying rates of diffusion was due partly to differences in proportion and properties of the molecularly dispersed gluten and partly to aggregates.All properties showed progressive changes both within and between the arbitrarily produced fractions. These results, therefore, support the hypothesis that gluten is a protein system showing progressive and regular changes in properties with change in solubility.


BioResources ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. 4137-4151 ◽  
Author(s):  
Aikfei Ang ◽  
Zaidon Ashaari ◽  
Edi Suhaimi Bakar ◽  
Nor Azowa Ibrahim

An alkali lignin (OL) with a weight-average molecular weight (Mw) of 11646 g/mol was used to prepare low-molecular weight lignin for resin synthesis. The low-molecular weight lignin feedstock was obtained via base-catalysed depolymerisation (BCD) treatments at different combined severity factors. Sequential fractionation of the OL and BCD-treated lignins using organic solvents with different Hildebrand solubility parameters were used to alter the homogeneity of the OL. The yield and properties of OL itself and OL and BCD-treated OL dissolved in propan-1-ol (F1), ethanol (F2), and methanol (F3) were determined. Regardless of the treatment applied, a small amount of OL was dissolved in F1 and F2. The BCD treatment did not increase the yield of F1 but did increase the yields of F2 and F3. Gel permeation chromatography (GPC) showed that the repolymerization reaction occurred in F3 for all BCD-treated OL, so these lignins were not suitable for use as feedstocks for resin production. The GPC, 13Carbon-nuclear magnetic resonance, and Fourier transform infrared spectroscopy analyses confirmed that the F3 in OL exhibited the optimum yield, molecular weight distribution, and chemical structure suitable for use as feedstocks for resin synthesis.


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