EP0220921A2
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Heat stable alkaline proteases produced by a bacillus
Assignee: Genex Corp
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Filing: 1986-10-21
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Granted: —
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Expires: 2006-10-21
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, Maryland as ATCC No. 53278.
Inventors
- 👤 Edmund J. Stellwag
- 👤 Wayne E. Swann
- 👤 David B. Stewart
- 👤 Donald R. Durham
- 👤 Carol A. Nolf
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 21:17
CPC Classifications
C; C12; C12N; C12N1/00; C12N1/20; C12N1/205; C11; C11D; C11D3/00; C11D3/16; C11D3/38; C11D3/386; 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: 23
Claim 1
Independent Claim
52 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 a Bacillus strain having the identifying characteristics of strain GX6638 in a fermentation medium and recovering therefrom the enzyme or enzymes produced by said Bacillus srain.
Claim 2
Dependent Claim
52 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 a Bacillus strain having the identifying characteristics of strain GX6638 in a fermentation medium and recovering therefrom the enzyme or enzymes produced by said Bacillus srain.
Claim 3
Dependent Claim
38 words
A process as claimed in claim 1, wherein following cultivation of the Bacillus strain the fermentation medium is centrifuged to remove cellular material, and then the enzyme(s) is/are concentrated in the supernatant, precipitated from said supernatant and recovered.
Claim 4
Dependent Claim
32 words
A member of the group consisting of Bacillus strain GX6638 (deposited at the American Type Culture Collection, Rockville, Maryland under accession No. ATCC 53278) and its proteolytic enzyme producing mutants and variants.
Claim 5
Dependent Claim
32 words
A member of the group consisting of Bacillus strain GX6638 (deposited at the American Type Culture Collection, Rockville, Maryland under accession No. ATCC 53278) and its proteolytic enzyme producing mutants and variants.
Claim 6
Dependent Claim
15 words
A biologically pure culture of a Bacillus strain having the identifying characteristics of strain GX6638.
Claim 7
Dependent Claim
15 words
A biologically pure culture of a Bacillus strain having the identifying characteristics of strain GX6638.
Claim 8
Dependent Claim
47 words
A mixture of enzymes producible by cultivation of Bacillus strain GX6638, 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 9
Dependent Claim
47 words
A mixture of enzymes producible by cultivation of Bacillus strain GX6638, 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
141 words
An alkaline protease characterized by the following properties: a) a molecular weight 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 is 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 11
Dependent Claim
141 words
An alkaline protease characterized by the following properties: a) a molecular weight 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 is 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
126 words
An 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 sulfanilamide azocasein is used as the substrate; d) an optimal activity at 70°c at a pH of about 10.5, said optimal activity measure against a synthetic peptide having the sequence succinylL-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-p-nitroanilide; e) a half-life of at least 200 minutes at 50°c and 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 13
Dependent Claim
126 words
An 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 sulfanilamide azocasein is used as the substrate; d) an optimal activity at 70°c at a pH of about 10.5, said optimal activity measure against a synthetic peptide having the sequence succinylL-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-p-nitroanilide; e) a half-life of at least 200 minutes at 50°c and 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
19 words
The alkaline protease of claim 6 or claim 7 produced by cultivation of Bacillus strain GX6638 under protease-producing conditions.
Claim 15
Dependent Claim
31 words
An alkaline protease producible by cultivation of Bacillus strain GX6638 under protease-producing conditions and characterized by a) a molecular weight of about 22,000 and b) an isoelectric point of about 9.5.
Claim 16
Dependent Claim
31 words
An alkaline protease producible by cultivation of Bacillus strain GX6638 under protease-producing conditions and characterized by a) a molecular weight of about 22,000 and b) an isoelectric point of about 9.5.
Claim 17
Dependent Claim
20 words
A washing composition containing an effective amount of the enzyme mixture of claim 5 having high alkaline and thermal stability.
Claim 18
Dependent Claim
22 words
A washing composition containing an effective amount of the enzyme of claim 6 or claim 7 having high alkaline and thermal stability.
Claim 19
Dependent Claim
16 words
A washing composition as claimed in claim 10 or claim 11 which further comprises a detergent.
Claim 20
Dependent Claim
20 words
A washing composition as claimed in claim 12 wherein said detergent is an ethoxylated alcohol or a linear alkyl sulfonate.
Claim 21
Dependent Claim
22 words
A washing composition as claimed in 12 or claim 13 which further comprises a detergent builder, foam booster, fragrance or coloring agent.
Claim 22
Dependent Claim
17 words
A washing composition as claimed in claim 14 wherein the detergent builder nitriloacetate, trisodium salt is present.
Claim 23
Independent Claim
32 words
A method of cleaning textiles which comprises washing said textiles in an aqueous solution comprising an effective amount of a washing composition as claimed in any one of claims 10 to 15.
References Cited (Backward Citations)
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