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US4480037A Expired / Lapsed US ⏱️ 0.0 years remaining

Alkaline protease and preparation method thereof

Assignee: Novozymes AS
•
Filing: 1983-02-08
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Granted: 1984-10-30
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Expires: 2003-02-08
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Abstract

Abstract Alkaline protease API-21 having the following physicochemical properties: (1) Activity: Hydrolyzes casein under alkaline conditions, optimum pH range is 10 to 11, and has optimum temperature range for activity of 45 DEG C. to 50 DEG C.; (2) Stability: No decrease in enzymatic activity observed up to 40 DEG C. upon heating without substrated for 10 minutes at a pH of 10, but 90% or more decrease of enzymatic activity observed at 50 DEG C. for 10 minutes at a pH of 10. Deactivation occurs at 50 DEG C. for 10 minutes at a pH of 8 and complete deactivation substantially occurs at 60 DEG C. for 10 minutes at a pH of 8. The protease is stable under alkaline conditions, especially within a pH range of 7 to 11.5, and is stable against freezing and freeze-drying; and (3) Enzyme molecule: The protease is protein having a molecular weight of about 22000 as estimated by a gel filtration method, an isoelectric point of 7.4 as estimated by an electrofocusing method, an absorption peak of 275 to 282 nm in a UV absorption spectrum, IR absorption characteristics as shown in FIG. 5, and a serine residue is the active site thereof. This alkaline protease is prepared by cultivating novel species of NKS-21 belonging to genus Bacillus.

Inventors

  • 👤 Fiji Ichishima
  • 👤 Takashi Onouchi

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-02 17:35
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/52; C12N9/54; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832

Claims Analysis

Total Claims: 4
Claim 1
Independent Claim 200 words
Alkaline protease API-21 having the following physicochemical properties: (1) Activity: Hydrolyzes casein under alkaline conditions, with an optimum pH range of 10 to 11, and has optimum temperature range for activity of 45° C. to 50° C.; (2) Stability: No decrease in enzymatic activity observed up to 40° C. upon heating without temperature substrate for 10 minutes at a pH of 10, but a 90% or more decrease of enzymatic activity observed at 50° C. for 10 minutes at a pH of 10. Deactivation occurs at 50° C. upon heating for 10 minutes at a pH of 8 and complete deactivation substantially occurs at 60° C. for 10 minutes at a pH of 8. The protease is stable under alkaline conditions, especially within a pH range of 7 to 11.5, and is stable against freezing and freeze-drying; and (3) Enzyme molecule: The protease is protein having a molecular weight of about 22000 as estimated by a gel filtration method, an isoelectric point of 7.4 as estimated by an electrofocusing method, an absorption peak of 275 to 282 nm in a UV absorption spectrum, IR absorption characteristics as shown in FIG. 5, and a serine residue in the active site thereof.
Claim 2
Dependent Claim 200 words
Alkaline protease API-21 having the following physicochemical properties: (1) Activity: Hydrolyzes casein under alkaline conditions, with an optimum pH range of 10 to 11, and has optimum temperature range for activity of 45° C. to 50° C.; (2) Stability: No decrease in enzymatic activity observed up to 40° C. upon heating without temperature substrate for 10 minutes at a pH of 10, but a 90% or more decrease of enzymatic activity observed at 50° C. for 10 minutes at a pH of 10. Deactivation occurs at 50° C. upon heating for 10 minutes at a pH of 8 and complete deactivation substantially occurs at 60° C. for 10 minutes at a pH of 8. The protease is stable under alkaline conditions, especially within a pH range of 7 to 11.5, and is stable against freezing and freeze-drying; and (3) Enzyme molecule: The protease is protein having a molecular weight of about 22000 as estimated by a gel filtration method, an isoelectric point of 7.4 as estimated by an electrofocusing method, an absorption peak of 275 to 282 nm in a UV absorption spectrum, IR absorption characteristics as shown in FIG. 5, and a serine residue in the active site thereof.
Claim 3
Independent Claim 40 words
A method for preparing alkaline protease API-21 comprising the steps of: (a) cultivating species of NKS-21 of genus Bacillus capable of producing alkaline protease API-21 in an alkaline culture medium; and (b) recovering alkaline protease API-21 from the cultivated product.
Claim 4
Independent Claim 40 words
A method for preparing alkaline protease API-21 comprising the steps of: (a) cultivating species of NKS-21 of genus Bacillus capable of producing alkaline protease API-21 in an alkaline culture medium; and (b) recovering alkaline protease API-21 from the cultivated product.

References Cited (Backward Citations)

US4052262A Cited
GB1263765A Cited
US3827938A Cited
US3871963A Cited
US3905869A Cited
US4002572A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1987-10-30 1988-04-30 1988-10-30 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1991-10-30 1992-04-30 1992-10-30 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1995-10-30 1996-04-30 1996-10-30 $7700.00 Window Closed / Lapsed

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