LINGCOD MUSCLE PHOSPHORIBOISOMERASE AND RIBULOSE 5′-PHOSPHATE 3′-EPIMERASE

1959 ◽  
Vol 37 (1) ◽  
pp. 961-973
Author(s):  
H. L. A. Tarr
Keyword(s):  
Acid Phosphatase ◽  
Ion Exchange ◽  
Ammonium Sulphate ◽  
Simple Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Water Extraction ◽  
Enzyme Preparations

Comparatively pure phosphoriboisomerase and ribulose 5′-phosphate 3′-epimerase enzyme preparations were obtained from lingcod muscle by a simple procedure involving water extraction, saturation of the extract with ammonium sulphate, dialysis, brief heating to 55 °C, lyophilization of the solution, and final separation by ion exchange chromatography, using diethylamiuoethyl cellulose columns. Both enzymes have broad pH optima above pH 7.0, but are rapidly inactivated below this value. The following equilibria were established and compared with those obtained by other investigators: [Formula: see text], 1.35:1.0; [Formula: see text], 1: 1.5 and [Formula: see text], 1:0.58:0.66. The ketopentulose phosphate resulting from the action of phosphoriboisomerase on D-ribose 5-phosphate was isolated and identified as D-ribulose5-phosphate. Both D-ribulose and D-xylulose were demonstrated after subjecting a product of epimerase action to hydrolysis by acid phosphatase and ion exchange chromatography.

LINGCOD MUSCLE PHOSPHORIBOISOMERASE AND RIBULOSE 5′-PHOSPHATE 3′-EPIMERASE

1959 ◽  
Vol 37 (8) ◽  
pp. 961-973 ◽  
Author(s):  
H. L. A. Tarr
Keyword(s):  
Acid Phosphatase ◽  
Ion Exchange ◽  
Ammonium Sulphate ◽  
Simple Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Water Extraction ◽  
Enzyme Preparations

Comparatively pure phosphoriboisomerase and ribulose 5′-phosphate 3′-epimerase enzyme preparations were obtained from lingcod muscle by a simple procedure involving water extraction, saturation of the extract with ammonium sulphate, dialysis, brief heating to 55 °C, lyophilization of the solution, and final separation by ion exchange chromatography, using diethylamiuoethyl cellulose columns. Both enzymes have broad pH optima above pH 7.0, but are rapidly inactivated below this value. The following equilibria were established and compared with those obtained by other investigators: [Formula: see text], 1.35:1.0; [Formula: see text], 1: 1.5 and [Formula: see text], 1:0.58:0.66. The ketopentulose phosphate resulting from the action of phosphoriboisomerase on D-ribose 5-phosphate was isolated and identified as D-ribulose5-phosphate. Both D-ribulose and D-xylulose were demonstrated after subjecting a product of epimerase action to hydrolysis by acid phosphatase and ion exchange chromatography.

scholarly journals The synthesis and purification of 2′-carboxy-d-arabinitol 1-phosphate, a natural inhibitor of ribulose 1,5-bisphosphate carboxylase, investigated by31P n.m.r

1989 ◽  
Vol 260 (3) ◽  
pp. 711-716 ◽  
Author(s):  
S Gutteridge ◽  
G S Reddy ◽  
G Lorimer
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Ion Exchange ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzymic Hydrolysis ◽  
Bisphosphate Carboxylase ◽  
Natural Inhibitor

2′-Carboxy-D-arabinitol 1-phosphate (2CA1P), a natural inhibitor of ribulose 1,5-bisphosphate carboxylase was synthesized from 2′-carboxy-D-arabinitol 1,5-bisphosphate (2CABP). The selective dephosphorylation of 2CABP with either acid phosphatase or alkaline phosphatase was investigated by using 31P n.m.r. The n.m.r. spectra of the progress of the reactions indicated that both phosphatases preferentially removed the 5-phosphate from the bisphosphate. After the consumption of all of the bisphosphate, alkaline phosphatase generated a mixture of 2′-carboxy-D-arabinitol 1- and 5-monophosphates in the ratio of about 4:1, along with Pi. The enzyme also hydrolysed the monophosphates to 2′-carboxyarabinitol, thus decreasing the yield of 2CA1P further. In contrast, acid phosphatase catalysed almost quantitative conversion of 2CABP into 2CA1P, preferring to hydrolyse only the 5-phosphate. In either case, separation of the 2CA1P from Pi or other products of enzymic hydrolysis was readily accomplished by conventional ion-exchange chromatography or h.p.l.c.

Two Different Chalcone Synthase Activities from Spinach

1985 ◽  
Vol 40 (3-4) ◽  
pp. 160-165 ◽  
Author(s):  
L. Beerhues ◽  
R. Wiermann
Keyword(s):  
Ion Exchange ◽  
Substrate Specificity ◽  
Chalcone Synthase ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzyme Preparations ◽  
Assay Mixture ◽  
Suitable Substrate ◽  
Malonyl Coa ◽  
Young Leaves

Chalcone synthase activity was found in enzyme preparations from spinach. In homogenates of young leaves two different activities of the enzyme could be separated by DEAE-ion exchange chromatography and chromatofocusing. Both activities formed naringenin with [2-14C] malonyl- CoA and 4-coumaroyl-CoA as substrates. They exhibited only slight differences in substrate specificity. For both activities 4-coumaroyl-CoA proved to be the most suitable substrate at both pH 6.8 and 8.0. Eriodictyol and homoeriodictyol formation from caffeoyl-CoA and feruloyl-CoA, respectively, only occured at pH 6.8. The formation of naringenin by the two activities was maximal at pH 7.5-8.0 and dependent upon the DTE-concentration in the assay mixture.

scholarly journals A simple procedure for the isolation of protein disulphide-isomerase

1987 ◽  
Vol 247 (1) ◽  
pp. 237-239 ◽  
Author(s):  
J Koivu ◽  
R Myllylä ◽  
K I Kivirikko
Keyword(s):  
Ion Exchange ◽  
Affinity Chromatography ◽  
Simple Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Beta Subunit ◽  
Protein Disulphide Isomerase ◽  
Step Procedure ◽  
Beta 2

A two-step procedure is described for the purification of protein disulphide-isomerase (PDI). This procedure is based on the previous finding that the beta-subunit of the prolyl 4-hydroxylase tetramer (alpha 2 beta 2) is identical with PDI [Koivu, Myllylä, Helaakoski, Pihlajaniemi, Tasanen & Kivirikko (1987) J. Biol. Chem. 262, 6447-6449; Pihlajaniemi, Helaakoski, Tasanen, Myllylä, Huhtala, Koivu & Kivirikko (1987) EMBO J. 6, 643-649]. The procedure involves purification of the prolyl 4-hydroxylase tetramer by a simple affinity chromatography and subsequent isolation of the beta-subunit from the dissociated tetramer by ion-exchange chromatography.

Studies on an Inhibitor of Plasminogen Activation in Human Serum

1973 ◽  
Vol 30 (02) ◽  
pp. 414-424 ◽  
Author(s):  
Ulla Hedner
Keyword(s):  
Ion Exchange ◽  
Human Serum ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Specific Adsorption ◽  
Partial Purification ◽  
Plasminogen Activation ◽  
Preparative Electrophoresis

SummaryA procedure is described for partial purification of an inhibitor of the activation of plasminogen by urokinase and streptokinase. The method involves specific adsorption of contammants, ion-exchange chromatography on DEAE-Sephadex, gel filtration on Sephadex G-200 and preparative electrophoresis. The inhibitor fraction contained no antiplasmin, no plasminogen, no α1-antitrypsin, no antithrombin-III and was shown not to be α2 M or inter-α-inhibitor. It contained traces of prothrombin and cerulo-plasmin. An antiserum against the inhibitor fraction capable of neutralising the inhibitor in serum was raised in rabbits.

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