Enzyme, its method of production and its application to the prep

Details Enzyme, its method of production and its application to the prep Enzyme, its method of production and its application to the prep

1986-10-21

1990-04-10

Warren, Charles F.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Enzymatic production of a protein or polypeptide

435183, 435859, C12P 2102, C12M 900, C12R 1265

Patent

active

049160621

ABSTRACT:
A polymeric enzyme having a molecular weight higher than or equal to 200,000 and not exhibiting any proteolytic activity on casein derived from a culture of Micrococcus caseolyticus is disclosed, as is a method of producing the enzyme by filtration on gel buffered with an aqueous buffer of an enzymatic system prepared by lyophilization of an aqueous concentrate formed by ultrafiltration of the solution obtained by dissolving, in an aqueous solution of calcium chloride, an enzymatic system precipitated from a supernatant of a micro-organism culture broth by ammonium sulphate, wherein the enzyme is not retained on a gel calibrated for proteins with a molecular weight of between 5,000 and 20,000, buffered with a 20 mM TRIS-HCl buffer, at pH 7.5, containing 1.5 mM calcium chloride, and the enzymatic system is derived from a culture broth of said Micrococcus caseolyticus. The enzyme may be used for the production of aspartame by condensation of L-aspartic acid with methyl DL-phenylalaninate.

REFERENCES:
patent: 4086139 (1978-04-01), Hoerle
patent: 4165311 (1979-08-01), Isowa et al.
patent: 4666838 (1987-05-01), Yokozeki et al.
Dezmazeaud et al., "Facteurs Intervenant dans la Production du Systeme Proteolytique Chez Micrococcus caseolyticus", Ann. Biol. Anim. Bioch. Biophys., 1968, 8, (3), pp. 419-429.
Dezmazeaud et al., "Isolement, Purification et Proprietes d'une Protease Exocellulaire de Micrococcus caseolyticus", Ann. Biol. Anim. Bioch. Biophys., 1968, 8 (4), pp. 505-577.
Mazur et al., "Structure--Taste Relationships of Some Dipeptides", J. Am. Chem. Soc., 91:10, May 7, 1969, pp. 2684-2691.
Dezmazeaud et al., "Specifite de la protease neutre de Micrococcus caseolyticus", Eur. J. Biochem., vol. 19 (1971), pp. 51-55.

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