PURIFICATION AND PARTIAL CHARACTERIZATION OF THERMOACTINOMYCES VULGARIS AMYLASES

1967 ◽  
Vol 13 (9) ◽  
pp. 1157-1163 ◽  
Author(s):  
M. J. Kuo ◽  
P. A. Hartman
Keyword(s):  
Ion Exchange ◽  
Column Chromatography ◽  
Maximum Rate ◽  
Amylase Activity ◽  
Enzyme Preparations ◽  
Ph Values ◽  
Ion Exchange Column ◽  
Optimum Ph ◽  
Thermoactinomyces Vulgaris

An α-amylase from Thermoactinomyces vulgaris has been purified about 100-fold. Its optimum pH was between 5.9 and 7.0, and the maximum rate was achieved at 60 °C. In the absence of substrate, the enzymes were more stable at pH 5.9 than at higher or lower pH values; inactivation was rapid at pH 7.0. Temperatures of 70 °C or greater also caused rapid denaturation of the enzyme in the absence of substrate. Three major peaks of amylase activity were detected when purified enzyme preparations were passed through Sephadex G-75 columns. At least two of these amylases were interconvertible. Four or five T. vulgaris proteinases also were separated, using ion exchange column chromatography.

scholarly journals PURIFICATION AND CHARACTERIZATION OF POLYGALACTURONASE OF `FUJI' APPLES.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 653e-653
Author(s):  
Seung-Ryeul Shin ◽  
Jae-Kyun Byun ◽  
Kyung-Ho Chang
Keyword(s):  
Ion Exchange ◽  
Column Chromatography ◽  
Malus Domestica ◽  
Gel Filtration ◽  
Reducing Sugar ◽  
Purification And Characterization ◽  
Stable Temperature ◽  
Ion Exchange Column ◽  
Optimum Ph

Polygalacturonase (PG) was purified from apple, Malus domestica Borkh, cv. Fuji by gel filtration, CM cellulose ion exchange column chromatography and characterized by means of several biochemical methods. Two forms of isozymes, PG-I and PG-II, were detected and the activities of PG-I were found to be higher than PG-II. The Km and Vmax values were calculated to be 1.54 mg/ml and 0.25 μM with reducing sugar 1 ml/30min., respectively. The PG was active between pH 3 and 8 with the optimum pH of about 4-5. The stable temperature for the PG was below 55°C with 30°C optimum. The PG activities were increased by Na* and Cu**, but were inhibited by Ag*, EDTA and SDS.

scholarly journals AMINO ACID COMPOSITION AND ELECTROPHORETIC PROPERTIES OF HYPERTENSIN I

1955 ◽  
Vol 102 (4) ◽  
pp. 435-440 ◽  
Author(s):  
Leonard T. Skeggs ◽  
Walton H. Marsh ◽  
Joseph R. Kahn ◽  
Norman P. Shumway
Keyword(s):  
Amino Acids ◽  
Quantitative Analysis ◽  
Amino Acid ◽  
Ion Exchange ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Isoelectric Point ◽  
Column Chromatography ◽  
Isoleucine Leucine ◽  
Ion Exchange Column

A preparation of hypertensin I was purified by countercurrent distribution and was shown to migrate as a single component in starch blocks at pH 9.3 and 4.2. It had an isoelectric point of 7.7. Quantitative analysis by ion exchange column chromatography showed eight amino acids in approximately unimolar proportion: aspartic, proline, valine, isoleucine, leucine, tyrosine, phenylalanine, and arginine. There were in addition two moles of histidine.

Quantitative Method for Estimating Fecal Amino Acids

1975 ◽  
Vol 21 (10) ◽  
pp. 1437-1440 ◽  
Author(s):  
Christopher W I Owens ◽  
Walfredo Padovan
Keyword(s):  
Amino Acids ◽  
Ion Exchange ◽  
Column Chromatography ◽  
Quantitative Method ◽  
Normal Values ◽  
Amino Nitrogen ◽  
Exchange Column ◽  
Ion Exchange Column ◽  
Fresh Stool

Abstract We describe a method for quantitatively estimating 24 ninhydrin-reacting substances, including the commoner amino acids, in fecal dialysate prepared from ingested dialysis bags retrieved from fresh stool. It is accurate to 2 µmol of α-amino nitrogen per liter of fecal dialysate, and for most substances recovery of added standards is 100%. It involves dilution, ultrafiltration, and automated ion-exchange column chromatography of the dialysate. Some normal values are provided.

Induced binding of proteins by ammonium sulfate in affinity and ion-exchange column chromatography

2007 ◽  
Vol 70 (3) ◽  
pp. 493-498 ◽  
Author(s):  
Tsutomu Arakawa ◽  
Kouhei Tsumoto ◽  
Daisuke Ejima ◽  
Yoshiko Kita ◽  
Yasushi Yonezawa ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Ammonium Sulfate ◽  
Column Chromatography ◽  
Exchange Column ◽  
Ion Exchange Column

scholarly journals Fractionnement sur échangeur anionique des colloïdes glucidiques solubles (gommes et pectines) du jus de raisin sain

OENO One ◽  
1980 ◽  
Vol 14 (1) ◽  
pp. 29
Author(s):  
Denis Dubourdieu ◽  
Pascal Ribéreau-Gayon
Keyword(s):  
Ion Exchange ◽  
Column Chromatography ◽  
Galacturonic Acid ◽  
Exchange Column ◽  
Ion Exchange Column ◽  
Neutral Sugars

<p style="text-align: justify;">Les gommes d'un moût de raisins sains sont fractionnés par chromatographie d'échange anionique sur D.E.A.E. Sephadex A 25. On peut ainsi caractériser des gommes neutres qui ne contiennent que des oses neutres dans leur molécule et des gommes acides qui comportent des oses neutres et de l'acide galacturonique.</p><p style="text-align: justify;">+++</p><p style="text-align: justify;">Gums of juice from healthy grappes are fractionnated by ion exchange column chromatography on D.E.A.E. Sephadex A 25. So are separated neutral gums, containing only neutral sugars in their molecules, from acidic gums, that contains neutral sugars and galacturonic acid.</p>

scholarly journals Column chromatography of human small-intestinal maltase, isomaltase and invertase activities

1969 ◽  
Vol 111 (2) ◽  
pp. 139-146 ◽  
Author(s):  
A. Dahlqvist ◽  
U. Telenius
Keyword(s):  
Ion Exchange ◽  
Column Chromatography ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Heat Inactivation ◽  
Enzyme Preparations ◽  
Small Intestinal ◽  
Two Peaks ◽  
Second Peak ◽  
Intestinal Disaccharidases

1. The maltase, isomaltase and invertase (sucrase) activities of solubilized mucosal preparations from human jejunum and ileum were studied with column chromatography on anion-exchange (diethylaminoethyl- and triethylaminoethyl-)cellulose and Sephadex G-200 gel. 2. On ion-exchange cellulose columns both kinds of enzyme preparations yielded two major disaccharidase peaks. The first peak contained maltase Ia (=isomaltase) and maltase Ib (=invertase). The second peak contained maltase II and maltase III. 3. On Sephadex G-200 gel columns jejunal preparations yielded the corresponding peaks as on ion-exchange columns, but the peaks appeared in the reverse order in the effluent. The ileal preparation studied yielded a single peak on gel columns, containing all the activities studied and eluted with the ‘void volume’. 4. Precipitation with ethanol did not affect the behaviour of the enzymes during ion-exchange chromatography. When gel filtration was performed after ethanol precipitation of the enzymes, however, two peaks were obtained also with the ileal preparation, and subfractionation of the invertase was obtained with both kinds of preparations. 5. The second peak from ion-exchange chromatograms, containing maltase II and maltase III, on concentration was found to have very weak isomaltase activity, probably exerted by these enzymes as such. This activity accounts for only about 1% of the total isomaltase activity of the mucosa. 6. The results support the concept of the specificity of the human small-intestinal disaccharidases previously described after heat-inactivation experiments. The subfractionation of the invertase that under certain conditions is seen on Sephadex G-200 columns appears most likely to be an artifact. Consequently the nomenclature for the human maltose-, isomaltose- and sucrose-splitting enzymes proposed by another research group after gel-filtration chromatography studies should be abandoned. It seems more logical to keep the nomenclature based on heat inactivation [maltase Ia (=isomaltase), maltase Ib (=invertase or sucrase), maltase II and maltase III] until increased knowledge about the specificity and structure of these enzymes makes possible a more rational nomenclature.

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