Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Oxidoreductase
Patent
1980-05-13
1982-07-27
Shapiro, Lionel M.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Oxidoreductase
435192, 435232, 435259, 435816, C12N 902, C12N 908, C12N 988
Patent
active
043418678
ABSTRACT:
Various enzymes, in particular Cu,Zn-superoxide dismutase (SOD), catalase and carbonic acid anhydrase, are recovered from blood by admixing wholly or partly isolated blood cells with ethanol or a homologous alcohol until a concentration of 10 to 70% by volume and allowing them to stand for hemolysis of the blood cells and denaturation of the hemoglobin, and then adding water up to the double volume or more, and removing the precipitate of cell residuals, hemoglobin and other denatured proteins from the suspension. Then the desired enzymes are isolated from the solution obtained.
In particular, SOD and catalase can both be isolated by chromatography of the solution at a pH of 4.7 to 5.5 on a cation exchange resin of the same polarity as SOD in the pH range used and elution of the resin with a buffer solution which has a pH in the range 4.7 to 7.5 and an ionic strength in the range 0.01 to 1.0 M, SOD being eluted at the lowest pH and/or the lowest ionic strength.
The process allows both the hemolysis of the blood cells and the precipitation of the hemoglobin to be carried out in one step while retaining the activity of the desired enzymes, so that they can be obtained in a high yield in a manner which lends itself to use on an industrial scale. Further, SOD and catalase may be isolated and purified in a single chromatography step.
REFERENCES:
patent: 3579495 (1971-05-01), Huber
patent: 3763137 (1973-10-01), Huber et al.
Ku et al., Journal of Pharmaceutical Science, 1974, vol. 63, No. 1, pp. 60-64.
Briggs et al., Biochimica et Biophysica Acta, vol. 537, pp. 100-109, (1978).
De Forende Bryggerier A/S
Shapiro Lionel M.
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