Patent EP0265832A2 successfully refreshed from Google Patents.
EP0265832A2
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EP
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Novel alkaline cellulases and a microorganism for producing the same
Assignee: Kao Corp
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Filing: 1987-10-21
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Granted: —
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Expires: 2007-10-21
Abstract
Abstract Novel alkaline cellulase K, CMCase I and CMCase II are obtained by isolation from a culture product of Bacillus sp KSM-635. These enzymes stably work in a wide range including an alkaline side, and their activity is shown even at low temperatures. Further, they have a strong resistance to surfce active agents, chelating agents and proteinases. Therefore, they can be effectively utilized not only as an additive for clothing detergents, but also as a biomass and in other fields.
Inventors
- 👤 Susumu Ito
- 👤 Tomokazu Sato
- 👤 Katsuya Ozaki
- 👤 Shitsuw Shikata
- 👤 Kikuhiko Okamoto
- 👤 Shigeo Inoue
- 👤 Kenzo Koike
- 👤 Yuichi Ota
- 👤 Akira Takei
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-05 17:31
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/24; C12N9/2402; C12N9/2405; C12N9/2434; C12N9/2437; C12Y; C12Y302/00; C12Y302/01; C12Y302/01004; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
239 words
Alkaline cellulase K having the following physical and chemical properties. (1) Activity Having a Cx enzymatic activity of acting on carboxymethyl cellulose along with a weak C₁ enzymatic activity and a weak beta-glucoxidase activity. (2) Specificity on Substrates Acting on carboxymethyl cellulose(CMC), crystalline cellulose, Avicell, cellobiose, and p-nitrophenyl cellobioside(PNPC). (3) Working pH and optimum pH Having a working pH in the range of 4 to 12 and an optimum pH in the range of 9 to 10. (4) Stable pH Having stable pH values of 4.5 to 10.5 and 6.8 to 10 when allowed to stand at 40°C for 10 minutes and 30 minutes, respectively. (5) Working temperature range and working optimum temperature Working in a wide temperature range of from 10 to 65°C with an optimum temperature being recognized at about 40°C. (6) Influences of chelating agents The activity not impeded with ethylenediamine tetraacetic acid (EDTA), ethyleneglycol-bis-(β-aminoethylether) N,N,Nʹ,Nʹʹ-tetraacetic acid (EGTA), N,N-bis(carboxymethyl)glycine (nitrilotriacetic acid) (NTA), sodium tripolyphosphate (STPP) and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of surface active agents such as sodium linear alkylbenzenesulfonates (LAS), sodium alkylsulfates (AS), sodium polyoxyethylene alkylsulfates (ES), sodium alphaolefinsulfonates (AOS), sodium alpha-sulfonated aliphatic acid esters (alpha-SFE), sodium alkylsulfonates (SAS), polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), and dimethyldialkylammonium chloride. (8) Influences of proteinases Having a strong resistance to proteinases. (9) Molecular weight (determined by gel chromatography) Having a maximum peak at 180,000 ± 10,000.
Claim 2
Dependent Claim
239 words
Alkaline cellulase K having the following physical and chemical properties. (1) Activity Having a Cx enzymatic activity of acting on carboxymethyl cellulose along with a weak C₁ enzymatic activity and a weak beta-glucoxidase activity. (2) Specificity on Substrates Acting on carboxymethyl cellulose(CMC), crystalline cellulose, Avicell, cellobiose, and p-nitrophenyl cellobioside(PNPC). (3) Working pH and optimum pH Having a working pH in the range of 4 to 12 and an optimum pH in the range of 9 to 10. (4) Stable pH Having stable pH values of 4.5 to 10.5 and 6.8 to 10 when allowed to stand at 40°C for 10 minutes and 30 minutes, respectively. (5) Working temperature range and working optimum temperature Working in a wide temperature range of from 10 to 65°C with an optimum temperature being recognized at about 40°C. (6) Influences of chelating agents The activity not impeded with ethylenediamine tetraacetic acid (EDTA), ethyleneglycol-bis-(β-aminoethylether) N,N,Nʹ,Nʹʹ-tetraacetic acid (EGTA), N,N-bis(carboxymethyl)glycine (nitrilotriacetic acid) (NTA), sodium tripolyphosphate (STPP) and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of surface active agents such as sodium linear alkylbenzenesulfonates (LAS), sodium alkylsulfates (AS), sodium polyoxyethylene alkylsulfates (ES), sodium alphaolefinsulfonates (AOS), sodium alpha-sulfonated aliphatic acid esters (alpha-SFE), sodium alkylsulfonates (SAS), polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), and dimethyldialkylammonium chloride. (8) Influences of proteinases Having a strong resistance to proteinases. (9) Molecular weight (determined by gel chromatography) Having a maximum peak at 180,000 ± 10,000.
Claim 3
Dependent Claim
20 words
Alkaline cellulase K according to Claim 1, which is obtained by isolation from a culture product of Bacillus sp KSM-635.
Claim 4
Independent Claim
30 words
A method for producing alkaline cellulase K which comprises cultivating alkaline cellulase K-producing bacteria belonging to the genus Bacillus , and collecting alkaline cellulase K from the resulting culture product.
Claim 5
Independent Claim
30 words
A method for producing alkaline cellulase K which comprises cultivating alkaline cellulase K-producing bacteria belonging to the genus Bacillus , and collecting alkaline cellulase K from the resulting culture product.
Claim 6
Dependent Claim
221 words
CMCase I having the following physicochemical properties. (1) Activity Having the Cx enzymatic activity of acting on CMC along with a weak C₁ enzymatic activity and a weak beta-glucosidase activity. (2) Specificity on substrates Acting on CMC, crystalline cellulose, Avicel, cellubiose, and PNPC. (3) Working pH and optimum pH Having a working pH in the range of 3 to 12.5 and an optimum pH in the range of 6 to 11.5. (4) Stable pH Not inactivated at a pH of 5 to 12 when allowed to stand at 30°C for 1 hour. (5) Working temperature and working optimum temperature Working in a temperature range of from 10 to 60°C with an optimum temperature being at 22 to 53°C. (6) Influences of chelating agents The activity not impeded with EDTA, EGTA, NTA, STPP and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of LAS, AS, ES, AOS, alpha-SFE, SAS, polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), dimethyldialkylammonium chloride, and taurocholic acid. (8) Resistance to proteinases Having a strong resistance to proteinases. (9) Molecular weight (determined by gel chromatography) Having a molecular weight of this enzyme of 145,000 ± 10,000. (10) UV absorption spectrum This enzyme has a maximum absorption at 280 nm and a shoulder absorption at 290 nm, thus sugar components being contained.
Claim 7
Dependent Claim
221 words
CMCase I having the following physicochemical properties. (1) Activity Having the Cx enzymatic activity of acting on CMC along with a weak C₁ enzymatic activity and a weak beta-glucosidase activity. (2) Specificity on substrates Acting on CMC, crystalline cellulose, Avicel, cellubiose, and PNPC. (3) Working pH and optimum pH Having a working pH in the range of 3 to 12.5 and an optimum pH in the range of 6 to 11.5. (4) Stable pH Not inactivated at a pH of 5 to 12 when allowed to stand at 30°C for 1 hour. (5) Working temperature and working optimum temperature Working in a temperature range of from 10 to 60°C with an optimum temperature being at 22 to 53°C. (6) Influences of chelating agents The activity not impeded with EDTA, EGTA, NTA, STPP and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of LAS, AS, ES, AOS, alpha-SFE, SAS, polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), dimethyldialkylammonium chloride, and taurocholic acid. (8) Resistance to proteinases Having a strong resistance to proteinases. (9) Molecular weight (determined by gel chromatography) Having a molecular weight of this enzyme of 145,000 ± 10,000. (10) UV absorption spectrum This enzyme has a maximum absorption at 280 nm and a shoulder absorption at 290 nm, thus sugar components being contained.
Claim 8
Dependent Claim
20 words
CMCase I according to Claim 4, which is obtained by isolation from a culture product of Bacillus sp KSM 635.
Claim 9
Dependent Claim
212 words
CMCase II having the following physical and chemical properties. (1) Activity Having the Cx enzymatic activity of acting on CMC along with a weak C₁ enzymatic activity and a weak beta-glucosidase activity. (2) Specificity on substrates Acting on CMC, crystalline cellulose, Avicel, cellubiose and PNPC. (3) Working pH and optimum pH Having working pH in the range of 3 to 12.5 and an optimum pH in the range of 6 to 11.5. (4) pH stability Not inactivated at a pH of 5 to 12 when allowed to stand at 30°C for 1 hour. (5) Working temperature and working optimum temperature Working in a temperature range of from 5 to 58°C with an optimum temperature being at 14 to 45°C. (6) Influences of chelating agents The activity not impeded with EDTA, EGTA, NTA, STPP and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of LAS, AS, ES, AOS, alpha-SFE, SAS, polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), and dimethyldialkylammonium chloride. (8) Resistance to proteinases Having a strong resistance to proteinases (9) Molecular weight (determined by gel chromatography) The molecular weight of CMCase II is 170,000 + 20,000. (10) UV absorption spectrum Having a maximum absorption at 280 nm and a shoulder absorption at 290 nm.
Claim 10
Dependent Claim
212 words
CMCase II having the following physical and chemical properties. (1) Activity Having the Cx enzymatic activity of acting on CMC along with a weak C₁ enzymatic activity and a weak beta-glucosidase activity. (2) Specificity on substrates Acting on CMC, crystalline cellulose, Avicel, cellubiose and PNPC. (3) Working pH and optimum pH Having working pH in the range of 3 to 12.5 and an optimum pH in the range of 6 to 11.5. (4) pH stability Not inactivated at a pH of 5 to 12 when allowed to stand at 30°C for 1 hour. (5) Working temperature and working optimum temperature Working in a temperature range of from 5 to 58°C with an optimum temperature being at 14 to 45°C. (6) Influences of chelating agents The activity not impeded with EDTA, EGTA, NTA, STPP and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of LAS, AS, ES, AOS, alpha-SFE, SAS, polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), and dimethyldialkylammonium chloride. (8) Resistance to proteinases Having a strong resistance to proteinases (9) Molecular weight (determined by gel chromatography) The molecular weight of CMCase II is 170,000 + 20,000. (10) UV absorption spectrum Having a maximum absorption at 280 nm and a shoulder absorption at 290 nm.
Claim 11
Dependent Claim
20 words
CMCase II according to Claim 6, which is obtained by isolation from a culture product of Bacillus sp KSM 635.
Claim 12
Dependent Claim
224 words
Bacteria capable of producing alkali cellulase K, belonging to the genus Bacillus and having the following enzymatic properties. (1) Activity Having a Cx enzymatic activity of acting on CMC along with a weak C₁ enzymatic activity and a weak beta-glucoxidase activity. (2) Specificity on Substrates Acting on CMC, crystalline cellulose, Avicel, cellubiose, and PNPC. (3) Working pH and optimum pH Having a working pH in the range of 4 to 12 and an optimum pH in the range of 9 to 10 (Fig. 3). (4) pH stability Having stable pH values of 4.5 to 10.5 and 6.8 to 10 when allowed to stand at 40°C for 10 minutes and 30 minutes, respectively (Fig. 4). (5) Working temperature range and working optimum temperature Working in a wide temperature range of from 10 to 65°C with an optimum temperature being recognized at about 40°C (Fig. 5). (6) Influences of chelating agents The activity not impeded with EDTA, EGTA, NTA, STPP and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of surface active agents such as LAS, AS, ES, AOS, alpha-SFE, SAS, polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), and dimethyldialkylammonium chloride. (8) Influences of proteinases Having a strong resistance to proteinases. (9) Molecular weight (determined by gel chromatography) Having a maximum peak at 180,000 ± 10,000 (Fig. 7).
Claim 13
Dependent Claim
224 words
Bacteria capable of producing alkali cellulase K, belonging to the genus Bacillus and having the following enzymatic properties. (1) Activity Having a Cx enzymatic activity of acting on CMC along with a weak C₁ enzymatic activity and a weak beta-glucoxidase activity. (2) Specificity on Substrates Acting on CMC, crystalline cellulose, Avicel, cellubiose, and PNPC. (3) Working pH and optimum pH Having a working pH in the range of 4 to 12 and an optimum pH in the range of 9 to 10 (Fig. 3). (4) pH stability Having stable pH values of 4.5 to 10.5 and 6.8 to 10 when allowed to stand at 40°C for 10 minutes and 30 minutes, respectively (Fig. 4). (5) Working temperature range and working optimum temperature Working in a wide temperature range of from 10 to 65°C with an optimum temperature being recognized at about 40°C (Fig. 5). (6) Influences of chelating agents The activity not impeded with EDTA, EGTA, NTA, STPP and zeolite. (7) Influences of surface active agents Undergoing little inhibition of activity by means of surface active agents such as LAS, AS, ES, AOS, alpha-SFE, SAS, polyoxyethylene secondary alkyl ethers, fatty acid salts (sodium salts), and dimethyldialkylammonium chloride. (8) Influences of proteinases Having a strong resistance to proteinases. (9) Molecular weight (determined by gel chromatography) Having a maximum peak at 180,000 ± 10,000 (Fig. 7).
References Cited (Backward Citations)
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