Patent US4416150A successfully refreshed from Google Patents.
Patent EP0251446A2 successfully refreshed from Google Patents.
EP0251446A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Non-human Carbonyl hydrolase mutants, DNA sequences and vectors encoding same and hosts transformed with said vectors
Assignee: Danisco US Inc
•
Filing: 1987-04-28
•
Granted: —
•
Expires: 2007-04-28
Abstract
Abstract Novel carbonyl hydrolase mutants derived from the amino acid sequence of naturally-occurring or recombinant non-human carbonyl hydrolases and DNA sequences encoding the same. The mutant carbonyl hydrolases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant carbonyl hydrolase to encode the substitution, insertion or deletion of one or more amino acids in the amino acid sequence of a precursor carbonyl hydrolase. Such mutants have one or more properties which are different than the same property of the precursor hydrolase.
Inventors
- 👤 James Allen Wells
- 👤 Brian C. Cunningham
- 👤 Robert Mark Caldwell
- 👤 Richard Ray Bott
- 👤 David Aaron Estell
- 👤 Scott Douglas Power
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 17:39
CPC Classifications
C; C11; C11D; C11D3/00; C11D3/16; C11D3/38; C11D3/386; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/52; C12N9/54
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
109 words
A carbonyl hydrolase mutant having at least one property which is substantially different from the same property of a precursor carbonyl hydrolase from which the amino acid sequence of said carbonyl hydrolase mutant is derived, said property being selected from the group consisting of thermal stability and alkaline stability wherein said precursor carbonyl hydrolase is selected from the group consisting of naturally occurring carbonyl hydrolases and recombinant carbonyl hydrolases and said carbonyl hydrolase mutant amino acid sequence is derived by a method selected from the group consisting of the substitution, deletion and insertion of at least one amino acid in said amino acid sequence of said precursor carbonyl hydrolase.
Claim 2
Dependent Claim
109 words
A carbonyl hydrolase mutant having at least one property which is substantially different from the same property of a precursor carbonyl hydrolase from which the amino acid sequence of said carbonyl hydrolase mutant is derived, said property being selected from the group consisting of thermal stability and alkaline stability wherein said precursor carbonyl hydrolase is selected from the group consisting of naturally occurring carbonyl hydrolases and recombinant carbonyl hydrolases and said carbonyl hydrolase mutant amino acid sequence is derived by a method selected from the group consisting of the substitution, deletion and insertion of at least one amino acid in said amino acid sequence of said precursor carbonyl hydrolase.
Claim 3
Dependent Claim
135 words
A carbonyl hydrolase mutant having at least one property which is substantially different from the same property of a precursor carbonyl hydrolase from which the amino acid sequence of said carbonyl hydrolase mutant is derived, said property being selected from the group consisting of oxidative stability, substrate specificity, catalytic activity, thermal stability, alkaline stability and pH activity profile wherein said precursor carbonyl hydrolase is selected from the group consisting of naturally occurring carbonyl hydrolases and recombinant carbonyl hydrolases and said carbonyl hydrolase mutant amino acid sequence is derived by a method selected from the group consisting of deletion and insertion of at least one amino acid in said amino acid sequence of said precursor carbonyl hydrolase and substitution of more than one amino acid residue of said amino acid sequence of said precursor carbonyl hydrolase.
Claim 4
Dependent Claim
135 words
A carbonyl hydrolase mutant having at least one property which is substantially different from the same property of a precursor carbonyl hydrolase from which the amino acid sequence of said carbonyl hydrolase mutant is derived, said property being selected from the group consisting of oxidative stability, substrate specificity, catalytic activity, thermal stability, alkaline stability and pH activity profile wherein said precursor carbonyl hydrolase is selected from the group consisting of naturally occurring carbonyl hydrolases and recombinant carbonyl hydrolases and said carbonyl hydrolase mutant amino acid sequence is derived by a method selected from the group consisting of deletion and insertion of at least one amino acid in said amino acid sequence of said precursor carbonyl hydrolase and substitution of more than one amino acid residue of said amino acid sequence of said precursor carbonyl hydrolase.
Claim 5
Dependent Claim
100 words
A carbonyl hydrolase mutant derived by the replacement of at least one amino acid residue of a precursor carbonyl hydrolase with a different amino acid, said one amino acid residue being selected from the group of amino acid residues of Bacillus amyloliquefaciens subtilisin consisting of Tyr2l, Thr22, Ser24, Ser33, Asp36, Ala45, Gly46, Ala48, Ser49, Met50, Asn77, Ser87, Lys94, Va195, Leu96, Tyr104, Ilel07, Gly110, Met124, Asn155, Glul56, Lys170, Tyr171, Pro172, Phe189, Asp197, Met199, Ser204, Lys213, Tyr217, Ser221, His67, Leu126, Leu135, Gly97, Asp99, Ser101, Gly102, G1u103, Leu126, Gly127, Glyl28, Prol29, Tyr214, Gly215, and equivalent amino acid residues in other precursor carbonyl hydrolases.
Claim 6
Dependent Claim
100 words
A carbonyl hydrolase mutant derived by the replacement of at least one amino acid residue of a precursor carbonyl hydrolase with a different amino acid, said one amino acid residue being selected from the group of amino acid residues of Bacillus amyloliquefaciens subtilisin consisting of Tyr2l, Thr22, Ser24, Ser33, Asp36, Ala45, Gly46, Ala48, Ser49, Met50, Asn77, Ser87, Lys94, Va195, Leu96, Tyr104, Ilel07, Gly110, Met124, Asn155, Glul56, Lys170, Tyr171, Pro172, Phe189, Asp197, Met199, Ser204, Lys213, Tyr217, Ser221, His67, Leu126, Leu135, Gly97, Asp99, Ser101, Gly102, G1u103, Leu126, Gly127, Glyl28, Prol29, Tyr214, Gly215, and equivalent amino acid residues in other precursor carbonyl hydrolases.
Claim 7
Dependent Claim
124 words
A carbonyl hydrolase mutant having an amino acid sequence derived from the amino acid sequence of a precursor carbonyl hydrolase by the substituion of a differnt amino acid for more than one amino acid residue of said amino acid sequence of said precursor carbonyl hydrolase, said amino acid residues being selected from the group of amino acid residues of Bacillus amyloliquefaciens subtilisin consisting of Tyr21, Thr22, Ser24, Asp32, Ser33, Asp36, Ala45, Gly46, Ala48, Ser49, Met50, Asn77, Ser87, Lys94, Va195, Leu96, Tyr104, Ile107, Gly110, Met124, Alal52, Asn-155, Glul56, Gly166, Gly169, Lys170, Tyr171, Pro172, Phe189, Asp197, Met199, Ser204, Lys213, Tyr217, Ser221, Met222, His67, Leu126, Leu135, Gly97, Asp99, Ser101, Gly102, Glu103, Leu126, Gly127, Gly128, Pro129, Tyr214, Gly215, and equivalent amino acid residues in other precursor carbonyl hydrolases.
Claim 8
Dependent Claim
124 words
A carbonyl hydrolase mutant having an amino acid sequence derived from the amino acid sequence of a precursor carbonyl hydrolase by the substituion of a differnt amino acid for more than one amino acid residue of said amino acid sequence of said precursor carbonyl hydrolase, said amino acid residues being selected from the group of amino acid residues of Bacillus amyloliquefaciens subtilisin consisting of Tyr21, Thr22, Ser24, Asp32, Ser33, Asp36, Ala45, Gly46, Ala48, Ser49, Met50, Asn77, Ser87, Lys94, Va195, Leu96, Tyr104, Ile107, Gly110, Met124, Alal52, Asn-155, Glul56, Gly166, Gly169, Lys170, Tyr171, Pro172, Phe189, Asp197, Met199, Ser204, Lys213, Tyr217, Ser221, Met222, His67, Leu126, Leu135, Gly97, Asp99, Ser101, Gly102, Glu103, Leu126, Gly127, Gly128, Pro129, Tyr214, Gly215, and equivalent amino acid residues in other precursor carbonyl hydrolases.
Claim 9
Dependent Claim
45 words
The mutant of Claim 4 wherein said combinations are selected from the group consisting of Thr22/Ser87, S er24/Ser87, Ala45/Ala48, Ser49/Lys94, Ser49/Va195, Met50/Va195, Met50/Glyl10, Met50/Met124, Met50/Met222, Met124/Met222, Glu156/Gly166, Glu156/Gly169, Gly166/Met222, Gly169/Met222, Tyr21/Thr22, Met50/Met124/Met222, Tyr21/Thr22/Ser87, Met50/Glul56/ Glyl66/Tyr217, Met50/Glu156/Tyr217, Glu156/Gly169/ Tyr217, Ile170/Lys213, Ser204/Lys213, Met50/Ilel07/ Lys213 and Ser24/Met50/Ile107/Glu156/Gly166/Gly169/ Ser204/Lys213/Gly215/Tyr217.
Claim 10
Dependent Claim
135 words
A carbonyl hydrolase mutant derived by the replacement of at least one amino acid residue of a precursor carbonyl hydrolase with a different amino acid, said one amino acid residue being selected from the group of amino acid residues of of Bacillus amyloliquefaciens subtilisin consisting of Tyr21, Thr22, Ser24, Asp32, Ser33, Asp36, Ala45, Gly46, Ala48, Ser49, Met50, Asn77, Ser87, Lys94, Val95, Leu96, Tyrl04, Ile107, GlyllO, Met124, Alal52, Asn-155, Glu156, , Gly166, Glyl69, Lys170, Tyr171, Pro172, Phe189, Asp197, Metl99, Ser204, Lys213, Tyr217, Ser221, Met222, His67, Leu126, Leu135, Gly97, Asp99, Ser101, Gly102, Glu103, Leu126, Gly127, Gly128, Pro129, Tyr214, Gly2l5, and equivalent amino acid residues in other precursor carbonyl hydrolases, wherein said at least one amino acid residue of said precursor carbonyl hydrolase is replaced with the amino acid residues listed in TABLE I and TABLE II herein.
Claim 11
Dependent Claim
135 words
A carbonyl hydrolase mutant derived by the replacement of at least one amino acid residue of a precursor carbonyl hydrolase with a different amino acid, said one amino acid residue being selected from the group of amino acid residues of of Bacillus amyloliquefaciens subtilisin consisting of Tyr21, Thr22, Ser24, Asp32, Ser33, Asp36, Ala45, Gly46, Ala48, Ser49, Met50, Asn77, Ser87, Lys94, Val95, Leu96, Tyrl04, Ile107, GlyllO, Met124, Alal52, Asn-155, Glu156, , Gly166, Glyl69, Lys170, Tyr171, Pro172, Phe189, Asp197, Metl99, Ser204, Lys213, Tyr217, Ser221, Met222, His67, Leu126, Leu135, Gly97, Asp99, Ser101, Gly102, Glu103, Leu126, Gly127, Gly128, Pro129, Tyr214, Gly2l5, and equivalent amino acid residues in other precursor carbonyl hydrolases, wherein said at least one amino acid residue of said precursor carbonyl hydrolase is replaced with the amino acid residues listed in TABLE I and TABLE II herein.
Claim 12
Dependent Claim
34 words
The mutant of Claim 6 wherein the amino acid replacing said at least one amino acid residue in said precursor carbonyl hydrolase is selectd from the replacement amino acids listed in TABLE I herein.
Claim 13
Dependent Claim
11 words
Mutant DNA sequence encoding the mutant of claims 1 through 7.
Claim 14
Dependent Claim
10 words
Expression vector containing the mutant DNA sequence of claim 8.
Claim 15
Dependent Claim
10 words
Host cell .transformed with the expression vector of Claim 9.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.