Oral hygiene enzymes and method for preparation
AssigneeMerck & Co., Inc.
InventorsThomas H. Stoudt, Karl H. Nollstadt
Life of this patent
Estimating the term…
Abstract
Enzymes called cariogenanases, elaborated by microorganisms are isolated and characterized as polysaccharases capable of degrading cariogenan, a newly discovered glucan of dental plaque composed of .alpha.-(1.fwdarw.3) and .alpha.-(1.fwdarw.2) linkages. The enzymes are isolable from culture broths of various organisms by protein precipitation methods, and have utility as an anti-dental plaque agent, either as the sole enzymatic agent or in combination with dextranases and/or plaque dispersing proteases.
Claims7
A dental treating composition comprising a dental treating carrier, dextranase, and an effective amount of an enzyme capable of hydrolyzing a streptococci derived dental plaque glucan having a majority of .alpha.-(1.fwdarw.3)glucosidic bonds.
A dental treating composition comprising a dental treating carrier, dextranase, protease and an effective amount of an enzyme capable of hydrolyzing a streptococci derived dental plaque glucan having a majority of .alpha.-(1.fwdarw.3)glucosidic bonds.
A process for preparing an enzyme capable of hydrolyzing a streptococci devised dental plaque glucan having a majority of .alpha.-(1-3)glucosidic bonds which comprises:
- (a) growing a microorganism selected from the the group consisting of a fungus, NRRL No. 5305;
- a Streptomyces sp., NRRL No. 5306;
- a Streptomyces sp., NRRL no. 5307;
- a Streptomyces sp., NRRL No. 5308;
- a Streptomyces sp., NRRL No. 5309;
- a Bacillus sp., NRRL No. B-5300;
- a Bacillus sp., NRRL No. B-5301;
- a Bacillus sp., NRRL No. B-5302 a Bacillus sp., NRRL No. B-5303;
- or a Corynebacterium sp., NRRL No. B-5310;
- in a suitable nutrient medium.
- (b) removing insolubles from the broth;
- and
- (c) isolating the protein from the clarified broth, which protein comprises the desired enzyme.
A process for preparing an enzyme capable of hydrolyzing a streptococci derived dental plaque glucan having a majority of .alpha.-(1-3)glucosidic bonds which comprises:
- (a) growing a microorganism selected from the group consisting of a fugus, NRRL No. 5305;
- a Streptomyces sp., NRRL No. 5306;
- a Streptomyces sp., NRRL No. 5307;
- a Streptomyces sp., NRRL No. 5308;
- a Streptomyces sp., NRRL No. 5309 a Bacillus sp., NRRL No. B-5300;
- a Bacillus sp., NRRL No. B-5301;
- a Bacillus sp., NRRL No. 5302;
- a Bacillus sp, B-5303;
- or a Corynebacterium sp., NRRL No. B-5310;
- in a suitable mutrient medium.
- (b) removing insolubles from the broth;
- (c) concentrating the clear solution;
- (d) precipitating by addition of acetone;
- (e) suspending the precipitate in a buffer and separating insolubles;
- (f) precipitating with ammonium sulfate;
- and
- (g) dialyzing the precipitate, which dialyzed precipitate comprises the desired enzyme.
A process for preparing an enzyme capable of hydrolyzing a streptococci derived dental plaque glucan having a majority of .alpha.-(1-3)glucosidic bonds which comprises:
- (a) growing Bacillus sp., NRRL No. B-5300 in a suitable nutrient medium containing cariogenan as the only carbohydrate source until available cariogenan has been consumed;
- (b) removing insolubles from the broth;
- (c) concentrating the clear solution about 10 fold.
- (d) adding acetone at about -60.degree. C. to a final concentration of about 60%;
- (e) dissolving the precipitate in a phosphate buffer of about pH 7 and separating insolubles;
- (f) adding ammonium sulfate to saturation;
- and
- (g) dialyzing the precipitate, which dialyzed precipitate comprises the desired enzyme.
Dependencies are inferred from the claim text.