US2009181875A1PendingUtilityA1
Subtilase variants
Est. expiryNov 6, 2022(expired)· nominal 20-yr term from priority
Inventors:Henriette DraborgPeter Kamp HansenMads Eskelund BjornvadMads Norregaard-MadsenMikael Mikkelsen
C11D 3/386C12N 9/54
64
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Claims
Abstract
The present invention relates to novel subtilase variants exhibiting alterations relative to the parent subtilase in one or more properties including: wash performance, thermal stability, storage stability or catalytic activity. The variants of the invention are suitable for use in, e.g., cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions.
Claims
exact text as granted — not AI-modified1 - 15 . (canceled)
16 . A variant of a parent subtilase comprising substitutions at positions 9 and 15, wherein the variant has protease activity and each position corresponds to a position of the amino acid sequence of SEQ ID NO: 1.
17 . The variant of claim 16 , wherein the substitutions are S9R+A15T.
18 . The variant of claim 16 , further comprising a substitution at position 68.
19 . The variant of claim 18 , wherein the substitution at position 68 is V68A.
20 . The variant of claim 17 , further comprising a substitution at position 68.
21 . The variant of claim 20 , wherein the substitution at position 68 is V68A.
22 . The variant of claim 16 , comprising a set of substitutions selected from the group consisting of:
S3T + S9R + P14H + A15M + R19L + S99SD + V139I
V4A + S9R + A15T + G97GV + H120D
S9L + A15T + T22TV + V139L + Q245F
S9L + A15T + T22TV + V139L + Q245F + L262S
S9R + A13V + A15T + I35V + N62D + Q245F
S9R + A15M + A16P + T22S + S99AD
S9R + A15T
S9R + A15T + A16P + G97GA + P131S + Q137D + N204S
S9R + A15T + G20A + G97GV + H120D + P131H
S9R + A15T + G20* + L21F + P52T + N62D + Q245R
S9R + A15T + G20* + L21F + G61E + *61aP + Q245R
S9R + A15T + G20* + L21F + *61aA + V68A + Q245R
S9R + A15T + G20* + L21F + *61aS + V68A + G160D + Q245R
S9R + A15T + G20* + L21F + N62D + Q245N
S9R + A15T + G20* + L21F + N62D + Q245R
S9R + A15T + G20* + L21F + N62D + Q245R + S259G
S9R + A15T + G20* + L21F + N62E + Q245R
S9R + A15T + G20* + L21F + *62aS + N218D + Q245R
S9R + A15T + G20* + L21F + *63aG + Q245R + N272V
S9R + A15T + L21LC + V139L + R186H + Q245M
S9R + A15T + L21LG + T22TV + V139L + N204D + Q245N
S9R + A15T + L21LP + T22TV + M119I + N218D + Q245I
S9R + A15T + L21LP + T22TY + V139L + G160D + Q245L
S9R + A15T + L21LW + G100S + V139L + Q245V
S9R + A15T + T22A + N62D
S9R + A15T + T22A + V139L + Q245E
S9R + A15T + T22G + V139L + Q245L
S9R + A15T + T22L + V139L + Q245V + A254S
S9R + A15T + T22TG + N62D + V139L + Q245G
S9R + A15T + T22TG + N62D + V139L + Q245S
S9R + A15T + T22TG + N62D + V139L + Q245V
S9R + A15T + T22TL + G61E + L96LG + Q137D + Q245R
S9R + A15T + T22TL + N62D + I107V + V139L + Q245W
S9R + A15T + T22TL + N62D + Q245W
S9R + A15T + T22TQ + S101P
S9R + A15T + T22TW + N204D + Q245I
S9R + A15T + S24P + G61E + A85T + P239S + Q245A
S9R + A15T + V28I + V68A + Q245R + N252A
S9R + A15T + T38S + A98R + S99C + G100S +
H120N + P131S + Q137H
S9R + A15T + G46D + V68A + N218D + Q245R
S9R + A15T + F50S + H120D + P131H
S9R + A15T + D60DG
S9R + A15T + G61E + V68A + A98S + S99G + Q245R
S9R + A15T + G61E + A85T + Q137H + Y209C + Q245G
S9R + A15T + G61E + A85T + N218D + P239S + Q245L
S9R + A15T + G61E + A85T + P239L + Q245C
S9R + A15T + G61E + A85T + P239S + Q245C
S9R + A15T + G61E + A85T + P239S + Q245V
S9R + A15T + G61E + A98S + S99M + Q245R
S9R + A15T + G61E + H120S + Q137D + V139L + N218D
S9R + A15T + G61GP + Q245L
S9R + A15T + N62D + H120N + P131T
S9R + A15T + N62D + Q245R + N252M
S9R + A15T + N62D + Q245W + N252S
S9R + A15T + N62D + Q245W + N252V
S9R + A15T + N62ND + V139L + Q245E
S9R + A15T + N62ND + V139L + N261D
S9R + A15T + N62NG + Q245T
S9R + A15T + N62S + H120N + P131T + N218D
S9R + A15T + V68A
S9R + A15T + V68A + A98G + S99I + K237R + Q245R
S9R + A15T + V68A + A98G + S99V + Q245R
S9R + A15T + V68A + A98L + S99G + Q245R
S9R + A15T + V68A + A98L + Q245R
S9R + A15T + V68A + A98M + S99G + Q245R + T274A
S9R + A15T + V68A + A98M + Q245R + N248D
S9R + A15T + V68A + S99G + Q245R + N261D
S9R + A15T + V68A + H120D + P131S + Q137H + Q245R
S9R + A15T + V68A + H120N + P131S + Q137H + Q245M
S9R + A15T + V68A + H120N + P131S + Q137H + Q245R
S9R + A15T + V68A + H120N + N218D + Q245R
S9R + A15T + V68A + Q137D
S9R + A15T + V68A + A174V + Q245R
S9R + A15T + V68A + A194T + Q245R + N252E
S9R + A15T + V68A + N218D + Q245R
S9R + A15T + V68A + Q245R
S9R + A15T + V68A + Q245R + N252S
S9R + A15T + N76S + L111V + P131H + Q137D
S9R + A15T + N77S + L96LG + H120D + P131Q
S9R + A15T + A88V + A98R + S99G + G100C +
H120N + P131S + Q137H
S9R + A15T + K94N + H120N + P131H
S9R + A15T + L96LG + H120D + P131H + R186L
S9R + A15T + L96LG + H120D + P131S + Q137E
S9R + A15T + L96LG + H120D + G160D
S9R + A15T + L96LG + H120N + P131H + Q137E
S9R + A15T + L96LG + H120N + P131S
S9R + A15T + L96LG + H120N + P131S + Q137H
S9R + A15T + L96LG + H120N + P131S + Q137H + M222S
S9R + A15T + L96LG + H120Q + Q137E
S9R + A15T + L96LG + S130*
S9R + A15T + L96LG + P131H + Q137D
S9R + A15T + L96LG + P131Q + Q137D
S9R + A15T + L96LG + P131T + Q137H
S9R + A15T + L96LG + Q137E + Y209H
S9R + A15T + G97GA + H120D + P131H + Q137E
S9R + A15T + G97GA + H120D + P131S + Q137E
S9R + A15T + G97GA + H120D + Q137D
S9R + A15T + G97GA + H120D + Q137H + M222V
S9R + A15T + G97GA + H120N + Q137D + N248D
S9R + A15T + G97GA + H120N + Q137E
S9R + A15T + G97GA + H120N + S212L
S9R + A15T + G97GA + H120Q + P131H + Q137E
S9R + A15T + G97GA + H120Y + Q137H
S9R + A15T + G97GA + Q137D
S9R + A15T + G97GA + Q137E
S9R + A15T + G97GA + Q137E + L262V
S9G + A15T + G97GA + Q137H
S9R + A15T + G97GA + Q137H + N218S
S9R + A15T + G97GA + R186C
S9R + A15T + G97GG + L111I + P131T + Q137H
S9R + A15T + G97GG + H120D + P131H + Q137H
S9R + A15T + G97GG + H120D + Q137E
S9R + A15T + G97GG + H120N + Q137D
S9R + A15T + G97GG + P131S + Q137H
S9R + A15T + G97GV + S106A + P131H
S9R + A15T + G97GV + H120D + Q137H
S9R + A15T + G97GV + H120E + Q137H
S9R + A15T + G97GV + H120N
S9R + A15T + G97GV + H120Q
S9R + A15T + G97GV + H120Q + Y263F
S9R + A15T + G97GV + P131H + Q137H
S9R + A15T + G97GV + P131S
S9R + A15T + G97GV + P131T + Q137H
S9R + A15T + G97GV + Q137H
S9R + A15T + A98C + G100S + H120N + P131S + Q137H
S9R + A15T + A98G + S99C + H120N + P131S + Q137H
S9R + A15T + A98S + S99G + G100S +
H120N + P131S + Q137H
S9R + A15T + A98S + G100S + H120N + P131S + Q137H
S9R + A13T + S99A + S99SD + P131F
S9R + A15T + S99C + H120N + P131S + Q137H + M222S
S9R + A15T + S99G + G100S + H120N + P131S + Q137H
S9R + A15T + G102S + M175T + Q245R + N252D
S9R + A15T + L111I + H120N + Q137E
S9R + A15T + L111I + Q137E
S9R + A15T + L111V + Q137E + G211D
S9R + A15T + H120D + Q137D
S9R + A15T + H120G + Q137E + N218D
S9R + A15T + H120N + P131T + N218D
S9R + A15T + H120N + P131T + N218D + N269T
S9R + A15T + H120Q + P131C + Q137H
S9R + A15T + H120R + Q137D + N173S
S9R + A15T + G127E + P131R + Q137H
S9R + A15T + S130P
S9R + A15T + P131H + S144P
S9R + A15T + Y167I + R170L
S9R + A15T + N218D
S9R + A15T + T224A
S9R + A15T + Q245F
S9R + A15T + Q245L
S9R + A15T + Q245S
S9R + A15T + Q245V and
S9R + A15T + T260M.
23 . The variant of claim 16 , wherein the parent subtilase belongs to the sub-group I-S1.
24 . The variant of claim 16 , wherein the parent subtilase belongs to the sub-group I-S2.
25 . The variant of claim 16 , wherein the parent subtilase is subtilisin 309.
26 . The variant of claim 16 , further comprising one or more of the modifications K27R, *36D, S56P, N62D, V68A, N76D, S87N, G97N, S99SE, S101G, S103A, V104A, V104I, V104N, V104Y, S106A, H120D, H120N, N123S, G159D, Y167A, R170S, R170L, A194P, N204D, V205I, Q206E, L217D, N218S, N218D, M222S, M222A, T224S, A232V, K235L, Q236H, Q245R, N248D, N252K, T274A, S101G+V104N, S87N+S101G+V104N, K27R+V104Y+N123S+T274A, N76D+S103A+V104I, S99D+S101R+S103A+V1041+G160S, S3T+V4I+S99D+S100R+S103A+V104I+G160S+V199M+V205I+L217D, S3T+V4I+S99D+S100R+S103A+V104I+G160S+A194P+V199M+V205I+L217D, S3T+V4I+S99D+S100R+S103A+V104I+G160S+V2051 and N76D+V104A.
27 . The variant of claim 16 , further comprising the following substitutions: S101G+S103A+V104I+G159D+A232V+Q236H+Q245R+N248D+N252K.
28 . A cleaning or detergent composition, comprising a variant of claim 16 and a surfactant.
29 . A composition of claim 28 , which additionally comprises one or more of an amylase, cellulase, cutinase, esterase, beta-galactosidase, glycoamylase, hemicellulase, lactase, ligninase, lipase, polygalacturonase, and protease.
30 . An isolated DNA sequence encoding a protease variant of claim 16 .
31 . An expression vector comprising the isolated DNA sequence of claim 30 .
32 . A microbial host cell transformed with the expression vector of claim 31 .
33 . A microbial host cell of claim 32 , which is a Bacillus.
34 . A method for producing a protease variant, comprising
(a) culturing a host of claim 32 under conditions conducive to the expression and secretion of the variant, and (b) recovering the protease variant.Join the waitlist — get patent alerts
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