Method of purifying glucose isomerase and a composition for stor

Details Method of purifying glucose isomerase and a composition for stor Method of purifying glucose isomerase and a composition for stor

1979-11-13

1980-12-02

Shapiro, Lionel M.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Isomerase

435814, 435816, 435827, 435886, 435890, 435853, C12N 992

Patent

active

042372313

ABSTRACT:
A process for purifying glucose isomerase comprises the steps of acid treatment and salt fractionation. An enzyme solution is treated with an acid, such as acetic acid, to a pH from about 3.5 to about 5.0. The proteinaceous solids are collected and extracted with a buffer, such as imidazole, whose solution has a pH of about 6 to about 8. The solution is then collected and a salt, such as ammonium sulfate, is dissolved therein from about 40% to about 50% of its saturation point. The proteinaceous solids which form are removed and additional ammonium sulfate is dissolved to attain from about 41% to about 60% of its saturation point, followed by collection of the solids containing purified enzyme. A composition which preserves enzyme activity upon storage of glucose isomerase and which imparts resistance to thermal deactivation of said enzyme comprises an aqueous solution of glycerol, a buffer whose solution is at a pH of about 6 to about 8, divalent cobalt ions and magnesium ions.

REFERENCES:
patent: 4077842 (1978-03-01), Cory
Palleroni et al., "Mannose Isomerase of Pseudomonas saccharophila, " Journal of Biological Chemistry, 1956, vol. 218, pp. 535-548.
Yamanaka, "D-Xylose Isomerase," Methods in Enzymology, vol. IX, pp. 588-593 (1966).
Takasaki et al., "Studies on Sugar Isomerizing Enzyme," Agr. Biol. Chem., vol. 33, No. 11, pp. 1527-1534 (1969).

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