Patent CA1076983A successfully refreshed from Google Patents.
US4980288A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Subtilisin with increased thermal stability
Assignee: Enzon Labs Inc
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Filing: 1987-12-14
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Granted: 1990-12-25
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Expires: 2007-12-14
Abstract
Abstract Cloned DNA is mutated by creating a single-stranded target region in a cloned DNA segment, and introducing a mutation into the single-stranded target region by treating the target region with a chemical or biological mutagenizing agent capable of introducing mutations into single-stranded DNA. The mutated target region then is rendered double-stranded and a microorganism is transformed with the mutated double-stranded DNA present in an expression vector. The transformed microorganism is cultivated under conditions wherein the mutated DNA is expressed to form an expression product, and the expression product is screened to identify a desired mutation in the DNA segment. Mutant subtilisins of enhanced thermal stability are also disclosed.
Inventors
- 👤 Philip N. Bryan
- 👤 Michele L. Rollence
- 👤 Michael W. Pantoliano
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 13:12
CPC Classifications
C; C11; C11D; C11D3/00; C11D3/16; C11D3/38; C11D3/386; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/102; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/52; C12N9/54; C12Q; C12Q1/00; C12Q1/34; C12Q1/37; C12Q1/68
Claims Analysis
Total Claims: 45
Claim 1
Independent Claim
25 words
A cloned mutant subtilisin gene coding for a subtilisin polypeptide comprising serine or aspartic acid substituted for asparagine at amino acid position 218 of subtilisin.
Claim 2
Dependent Claim
25 words
A cloned mutant subtilisin gene coding for a subtilisin polypeptide comprising serine or aspartic acid substituted for asparagine at amino acid position 218 of subtilisin.
Claim 3
Dependent Claim
34 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 4
Dependent Claim
34 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 5
Dependent Claim
33 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and alanine substituted for threonine at amino acid position 254 of subtilisin.
Claim 6
Dependent Claim
33 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and alanine substituted for threonine at amino acid position 254 of subtilisin.
Claim 7
Dependent Claim
31 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and serine substituted for glycine at amino acid position 166.
Claim 8
Dependent Claim
31 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and serine substituted for glycine at amino acid position 166.
Claim 9
Dependent Claim
44 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 and alanine substituted for threonine at amino acid position 254 of subtilisin.
Claim 10
Dependent Claim
44 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 and alanine substituted for threonine at amino acid position 254 of subtilisin.
Claim 11
Dependent Claim
24 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 12
Dependent Claim
24 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 13
Dependent Claim
23 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising proline substituted for serine at amino acid position 188 of subtilisin.
Claim 14
Dependent Claim
23 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising proline substituted for serine at amino acid position 188 of subtilisin.
Claim 15
Dependent Claim
22 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising glutamic acid substituted for alanine at amino acid position 116.
Claim 16
Dependent Claim
22 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising glutamic acid substituted for alanine at amino acid position 116.
Claim 17
Dependent Claim
23 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising isoleucine substituted for leucine at amino acid position 126 of subtilisin.
Claim 18
Dependent Claim
23 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide comprising isoleucine substituted for leucine at amino acid position 126 of subtilisin.
Claim 19
Dependent Claim
23 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide having threonine substituted for serine at amino acid position 53 of subtilisin.
Claim 20
Dependent Claim
23 words
A cloned mutant subtilisin gene coding for a mutant subtilisin polypeptide having threonine substituted for serine at amino acid position 53 of subtilisin.
Claim 21
Dependent Claim
19 words
A mutant substilisin polypepetide comprising serine or aspartic acid substituted for asparagine at amino acid position 218 of substilisin.
Claim 22
Dependent Claim
19 words
A mutant substilisin polypepetide comprising serine or aspartic acid substituted for asparagine at amino acid position 218 of substilisin.
Claim 23
Dependent Claim
27 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 24
Dependent Claim
27 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 25
Dependent Claim
26 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and alanine substituted for threonine at amino acid position 254 of subtilisin
Claim 26
Dependent Claim
26 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and alanine substituted for threonine at amino acid position 254 of subtilisin
Claim 27
Dependent Claim
24 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and serine substituted for glycine at amino acid position 166.
Claim 28
Dependent Claim
24 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and serine substituted for glycine at amino acid position 166.
Claim 29
Dependent Claim
37 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 and alanine substituted for threonine at amino acid position 254 of subtilisin.
Claim 30
Dependent Claim
37 words
A mutant subtilisin polypeptide having serine substituted for asparagine at amino acid position 218 and aspartic acid substituted for glycine at amino acid position 131 and alanine substituted for threonine at amino acid position 254 of subtilisin.
Claim 31
Dependent Claim
17 words
A mutant subtilisin polypeptide having aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 32
Dependent Claim
17 words
A mutant subtilisin polypeptide having aspartic acid substituted for glycine at amino acid position 131 of subtilisin.
Claim 33
Dependent Claim
16 words
A mutant subtilisin polypeptide having proline substituted for serine at amino acid position 188 of substilisin.
Claim 34
Dependent Claim
16 words
A mutant subtilisin polypeptide having proline substituted for serine at amino acid position 188 of substilisin.
Claim 35
Dependent Claim
15 words
A mutant substilisin polypeptide having glutamic acid substituted for alanine at amino acid position 116.
Claim 36
Dependent Claim
15 words
A mutant substilisin polypeptide having glutamic acid substituted for alanine at amino acid position 116.
Claim 37
Dependent Claim
16 words
A mutant subtilisin polypeptide having isoleucine substituted for leucine at amino acid position 126 of subtilisin.
Claim 38
Dependent Claim
16 words
A mutant subtilisin polypeptide having isoleucine substituted for leucine at amino acid position 126 of subtilisin.
Claim 39
Dependent Claim
16 words
A mutatnt subtilisin polypeptide having threonine substituted for serine at amino acid position 53 of subtilisin.
Claim 40
Dependent Claim
16 words
A mutatnt subtilisin polypeptide having threonine substituted for serine at amino acid position 53 of subtilisin.
Claim 41
Dependent Claim
17 words
The mutant subtilisin polypeptide of claim 15 produced by cultivating B. subtilis strain GX7150 (ATCC No. 53459).
Claim 42
Dependent Claim
7 words
B. subtilis strain GX7150 (ATCC No. 53459).
Claim 43
Dependent Claim
7 words
B. subtilis strain GX7150 (ATCC No. 53459).
Claim 44
Dependent Claim
18 words
A B. Subtilisin strain comprising the mutant gene of any one of claims 1 or 2 to 10.
Claim 45
Dependent Claim
17 words
A washing preparation comprising the subtilisin polypeptide of any one of claims 11 or 12 to 21.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-12-24 | 1994-06-25 | 1994-12-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-12-24 | 1998-06-25 | 1998-12-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-12-24 | 2002-06-25 | 2002-12-25 | $7700.00 | Window Closed / Lapsed |