US4771003A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Heat stable alkaline proteases produced by a bacillus
Assignee: Genex Corp
•
Filing: 1985-10-22
•
Granted: 1988-09-13
•
Expires: 2005-10-22
Abstract
Abstract Novel enzymes exhibiting proteolytic activity in alkaline media and stability at high temperatures and under alkaline conditions are produced by a novel Bacillus strain designated GX6638 or its mutants or variants. These enzymes are especially well-suited for inclusion in washing compositions. A culture of GX6638 has been deposited with the American Type Culture Collection, Rockville, Md. as ATCC No. 53278.
Inventors
- 👤 Edmund J. Stellwag
- 👤 Donald R. Durham
- 👤 Wayne E. Swann
- 👤 Carol A. Nolf
- 👤 David B. Stewart
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 10:31
CPC Classifications
C; C11; C11D; C11D3/00; C11D3/16; C11D3/38; C11D3/386; C12; C12N; C12N1/00; C12N1/20; C12N1/205; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/52; C12N9/54; C12R; C12R2001/00; C12R2001/01; C12R2001/07; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
54 words
A process for the preparation of one or more enzymes having proteolytic activity in alkaline media and exhibiting high alkaline and thermal stability which comprises cultivating Bacillus strain GX6638 or an alkaline protease producing mutant or variant thereof in a fermentation medium and recovering therefrom the enzyme or enzymes produced by said Bacillus strain.
Claim 2
Dependent Claim
54 words
A process for the preparation of one or more enzymes having proteolytic activity in alkaline media and exhibiting high alkaline and thermal stability which comprises cultivating Bacillus strain GX6638 or an alkaline protease producing mutant or variant thereof in a fermentation medium and recovering therefrom the enzyme or enzymes produced by said Bacillus strain.
Claim 3
Dependent Claim
19 words
The process of claim 1, wherein the fermentation medium comprises carbon, nitrogen, magnesium salts, calcium salts and trace elements.
Claim 4
Dependent Claim
37 words
The process of claim 1, wherein following cultivation of the Bacillus strain the fermentation medium is centrifuged to remove cellular material, then the enzyme or enzymes is concentrated in the supernatant, precipitated from said supernatant and recovered.
Claim 5
Dependent Claim
19 words
A member of the group consisting of Bacillus novel species GX6638 and its proteolytic enzyme producing mutants and variants.
Claim 6
Dependent Claim
19 words
A member of the group consisting of Bacillus novel species GX6638 and its proteolytic enzyme producing mutants and variants.
Claim 7
Dependent Claim
17 words
A biologically pure culture of Bacillus strain GX6638 or an alkaline protease producing mutant or variant thereof.
Claim 8
Dependent Claim
17 words
A biologically pure culture of Bacillus strain GX6638 or an alkaline protease producing mutant or variant thereof.
Claim 9
Dependent Claim
57 words
A substantially pure mixture of enzymes produced by cultivation. Bacillus strain GX6638 or an alkaline protease producing mutant or variant thereof, said mixture comprising at least three separate enzymes having proteolytic activity, wherein said three enzymes have molecular weights of about 22,000, about 27,500 and about 36,000, and isoelectric points of about 9.5, 5.2 and 4.2, respectively.
Claim 10
Dependent Claim
57 words
A substantially pure mixture of enzymes produced by cultivation. Bacillus strain GX6638 or an alkaline protease producing mutant or variant thereof, said mixture comprising at least three separate enzymes having proteolytic activity, wherein said three enzymes have molecular weights of about 22,000, about 27,500 and about 36,000, and isoelectric points of about 9.5, 5.2 and 4.2, respectively.
Claim 11
Dependent Claim
146 words
A substantially pure alkaline protease characterized by the following properties: (a) a molecular weight of about 27,500; (b) an isoelectric point of about 5.2; (c) an optimal proteolytic activity at a pH of about 9.5 at a temperature of 40°-50° C., said optimal activity measured in an assay wherein sulfanilamide azocasein was used as the substrate; (d) an optimal activity at 60° C. at a pH of about 10.5, said optimal activity measured against a synthetic peptide having the sequence succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-p-nitroanilide; (e) a half-life of about 9 minutes at 50° C. and a pH of about 10.5; (f) a half-life of about 1.8 minutes at 60° C. and a pH of about 10.5; (g) a half-life of about 14 minutes at 40° C. and a pH of about 12; and (h) a half-life of about 42 minutes at 40° C. and a pH of about 10.8.
Claim 12
Dependent Claim
146 words
A substantially pure alkaline protease characterized by the following properties: (a) a molecular weight of about 27,500; (b) an isoelectric point of about 5.2; (c) an optimal proteolytic activity at a pH of about 9.5 at a temperature of 40°-50° C., said optimal activity measured in an assay wherein sulfanilamide azocasein was used as the substrate; (d) an optimal activity at 60° C. at a pH of about 10.5, said optimal activity measured against a synthetic peptide having the sequence succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-p-nitroanilide; (e) a half-life of about 9 minutes at 50° C. and a pH of about 10.5; (f) a half-life of about 1.8 minutes at 60° C. and a pH of about 10.5; (g) a half-life of about 14 minutes at 40° C. and a pH of about 12; and (h) a half-life of about 42 minutes at 40° C. and a pH of about 10.8.
Claim 13
Dependent Claim
132 words
A substantially pure alkaline protease characterized by the following properties: (a) a molecular weight of about 36,000; (b) an isoelectric point of about 4.2; (c) an optimal proteolytic activity at a pH of about 9.5 at a temperature of about 40°-50° C., said optimal activity measured in an assay wherein sulfanilamideazocasein was used as the substrate; (d) an optimal activity at 70° C. at a pH of about 10.5, said optimal activity measured against a synthetic peptide having the sequence succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-p-nitroanilide; (e) a half-life of at least 200 minutes at 50° C. at a pH of 10.5; (f) a half-life of about 25 minutes at 60° C. and a pH of 10.5; and (g) a half-life of at least 420 minutes at a pH of 10.8, and a temperature of 40° C.
Claim 14
Dependent Claim
132 words
A substantially pure alkaline protease characterized by the following properties: (a) a molecular weight of about 36,000; (b) an isoelectric point of about 4.2; (c) an optimal proteolytic activity at a pH of about 9.5 at a temperature of about 40°-50° C., said optimal activity measured in an assay wherein sulfanilamideazocasein was used as the substrate; (d) an optimal activity at 70° C. at a pH of about 10.5, said optimal activity measured against a synthetic peptide having the sequence succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-p-nitroanilide; (e) a half-life of at least 200 minutes at 50° C. at a pH of 10.5; (f) a half-life of about 25 minutes at 60° C. and a pH of 10.5; and (g) a half-life of at least 420 minutes at a pH of 10.8, and a temperature of 40° C.
Claim 15
Dependent Claim
20 words
A washing composition containing an effective amount of the enzyme mixture of claim 6 having high alkaline and thermal stability.
Claim 16
Dependent Claim
19 words
A washing composition containing an effective amount of the enzyme of claim 9 having high alkaline and thermal stability.
Claim 17
Dependent Claim
13 words
The washing composition of claim 9 or 10, which further comprises a detergent.
Claim 18
Dependent Claim
18 words
The washing composition of claim 11, which further comprises a detergent builder, foam booster, fragrance or coloring agent.
Claim 19
Dependent Claim
18 words
The washing composition of claim 11 wherein said detergent is an ethoxylated alcohol or a linear alkyl sulfonate.
Claim 20
Dependent Claim
14 words
The washing composition of claim 12 wherein the detergent builder is nitriloacetate, trisodium salt.
References Cited (Backward Citations)
US3723250A
Cited
GB1240058A
Cited
US3790482A
Cited
US3931034A
Cited
GB1247292A
Cited
US4052262A
Cited
US4252663A
Cited
US3655513A
Cited
US3827938A
Cited
US3855064A
Cited
US3871963A
Cited
US4287101A
Cited
US4480037A
Cited
US4529525A
Cited
US4429044A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-09-13 | 1992-03-14 | 1992-09-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-09-13 | 1996-03-14 | 1996-09-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-09-13 | 2000-03-14 | 2000-09-13 | $7700.00 | Window Closed / Lapsed |