US 12,247,237 B2
Polypeptide variants
Lars Henrik Oestergaard, Charlottelund (DK); Annette Helle Johansen, Bronshoj (DK); Steffen Danielsen, Stenlose (DK); Roland Alexander Pache, Valby (DK); Johanne Moerch Jensen, Brighton (AU); Frank Winther Rasmussen, Roskilde (DK); Martin Gudmand, Holte (DK); and Kasper Damgaard Tidemand, Copenhagen (DK)
Assigned to Novozymes A/S, Bagsvaerd (DK)
Appl. No. 17/437,114
Filed by Novozymes A/S, Bagsvaerd (DK)
PCT Filed Apr. 6, 2020, PCT No. PCT/EP2020/059712
§ 371(c)(1), (2) Date Sep. 8, 2021,
PCT Pub. No. WO2020/207944, PCT Pub. Date Oct. 15, 2020.
Claims priority of application No. 19168512 (EP), filed on Apr. 10, 2019.
Prior Publication US 2022/0364138 A1, Nov. 17, 2022
Int. Cl. C12P 21/02 (2006.01); C11D 3/386 (2006.01); C12N 1/20 (2006.01); C12N 9/24 (2006.01)
CPC C12P 21/02 (2013.01) [C11D 3/38636 (2013.01); C12N 1/20 (2013.01); C12N 9/2402 (2013.01); C12Y 302/01052 (2013.01)] 20 Claims
 
1. A dispersin variant, comprising one or more substitutions selected from the group consisting of:
D2A, D2L, D2N, D2R, D2V, or D2W,
Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, or Q3T,
S12A,
H15F or H15Y,
T17W, T17C, T17E, T17F, T17M, T17R, or T17V,
V18L,
E19D, E19K, E19N, or E19P,
K22A, K22M, or K22V,
S23A, S23C, S23E, S23I, S23L, S23R, S23T, or S23V,
L24I,
V25R,
D26M,
Y30A, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, or Y30V,
G32L, G32M, or G32R,
N34D,
N43H or N43L,
N45D, N45L, or N45V,
A49W or A49Y,
Y52M,
G54L, G54M, or G54N,
S56T, S56W, or S57W,
E58N,
N59A, N59C, N59D, N59E, N59F, N59M, N59R, N59V, or N59W,
T60V,
N62C, N62D, N62H, N62Q, or N62W,
T63C, T63D, T63L, T63N, T63R, or T63V,
K67A or K67L,
N68L or N68Q,
L71H, L71N, L71R, L71V, or L71W,
S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, or S72Y,
I74L,
S77A,
D79V,
K80E, K80H, K80L, K80N, K80Q, K80R, K80V, or K80W,
D81A, D81G, D81L, D81N, D81R, D81S, D81T, D81V, or D81W,
I82V,
L90F,
E99Q or E99R,
L100S,
K103A, K103R, or K103V,
K104N or K104W,
D105N,
V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106R, V106W, or V106Y,
K107A, K107C, K107L, K107M, K107T, K107V, or K107W,
N110M, N110R, or N110V,
D111A, D111E, D111M, D111N, D111Q, D111R, D111V, or D111W,
V113T,
T114C or T114S,
Y116D, Y116N, or Y116R,
S117D, S117H, S117N, or S117P,
E118A, E118D, E118G, or E118L,
E119G or E119W,
T120I, T120L, T120M, T120V, or T120W,
D122H or D122R,
Y123W,
Y124C, Y124H, Y1241, Y124K, Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, or Y124W,
D125C, D125G, D125H, D125K, D125Q, or D125R,
N126V,
R127D, R127H, R127K, R127L, R127M, R127Q, or R127W,
V128A, V128C, V128D, V128L, or V128T,
D131V,
Q135A, Q135D, Q135E, Q135K, Q135M, or Q135Y,
D138K, D138L, D138M, D138N, D138Q, D138R, D138S, D138V, or D138W,
E139W,
V140I,
D142R or D142W,
Y145H, Y145L, Y145N, or Y145V,
P147A, P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S, P147T, P147V,
K148A, K148D, K148L, or K148V,
F149L, F149M, or F149N,
E150A, E150D, E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W, E150Y,
G151A, G151C, G151D, G151L, G151N, G151P, G151S, or G151W,
K152D, K152L, or K152R,
S163P,
G164D, G164E, G164H, G164S, or G164V,
V167A, V167D, V167E, V167L, V167P, V167Q, V167R, or V167W,
H168N,
L170A, L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V, L170W, or L170Y,
D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R, D171V, D171W, or D171Y,
I173C,
D174H, D174M, D174N, D174V, or D174W,
F175Y,
N177M,
Q178A, Q178K, Q178R, or Q178W,
I179T,
S181C, S181D, S181F, S181G, S181K, S181N, S181P, S181Q, S181T, S181V, or S181W,
E185A, E185M, E185R, E185V, or E185W,
S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q, S186R, S186V, or S186W,
K187C, K187D, K187G, K187R, K187S, K187V, or K187W,
Y188P,
E189L, E189V, or E189W,
S199C, S199L, S199M, or S199Y,
E200D, E200F, E200K, E200L, E200M, E200N, E200R, or E200W,
A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q, A203R, A203S, A203T, A203V, or A203W,
N204L, N204M, N204V, N204W, or N204Y,
L205I,
D207A, D207C, D207E, D207G, D207K, D207N, D207Q, D207R, D207S, D207V, or D207W,
S208A, S208C, S208D, S208G, S208L, S208Q, S208T, S208V, or S208W,
S210T,
Q215K, Q215R, Q215M, or Q215L,
S217V,
T218A, T218L, T218Q, T218R, or T218V,
S221N,
G222D,
E224A or E224P,
S225G,
N227A, N227Q, N227R, N227S, N227T, or N227K,
D230N, D230R, D230T, or D230W,
E232D or E232V,
N233D, N233E, N233H, N233Q, N233R, or N233W,
W234R,
G235A, G235W, G235E, G235F, G235H, G235I, G235L, G235M, G235N, G235P, G235S, or G235V,
S237C, S237G, S237M, S237N, S237W, or S237Y,
Y244C, Y244E, or Y244M,
L249H, L249K, L249Q, L249R, L249W, or L249Y,
S251A, S251L, S251N, S251R, or S251W,
N252P or N252C,
G253D or G253W,
F2541, F254L, F254M, or F254Y,
Q256D, Q256E, or Q256R,
N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R, N260T, N260V, N260W, or N260Y,
E261A, E261D, E261R, or E261W,
Q262F, Q262H, Q262W, or Q262Y,
M263K, M263L, or M263Q,
D264C, D264E, or D264N,
Y265F,
N267S or N267T,
W268C, W268E, W268M, or W268R,
Y270F,
A271D or A271G,
H272D, H272I, H272M, H272P, H272V, or H272W,
N273W,
K274R, K274A, or K274H,
F276A, F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V, or F276W,
I278A, I278K, 1278N, 1278Q, or I278V,
S279C, S279D, S279E, S279G, or S279N,
D280C or D280E,
Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, or Y281R,
Y282E or Y282N,
H283I,
A284I, A284L, A284N, A284P, A284T, or A284V,
T287N,
S288P, S288D, S288K, or S288N,
V290I,
K291L, K291R, or K291V,
T296C,
E300A or E300D,
H301C, H301N, or H301R,
T303A, T303C, T303G, T303K, T303Q, T303R, or T303W,
D304C or D304M,
L305M or L305N,
S306C,
K308A, K308D, K308E, K308G, K308I, K308L, K308Q, K308S, K308T, K308V, or K308Y,
K309A, K309C, K309D, K309E, K309G, K309H, K309L, K309M, K309N, K309Q, K309S, K309T, or K309I,
K312A, K312E, K312L, K312M, K312N, K312Q, K312S, or K312W,
E314I, E314L, or E314V,
L315I or L315V,
R319A,
Y321F and
N323R,
wherein the dispersin variant has at least 80% sequence identity to SEQ ID NO: 1, hexosaminidase activity, and increased stability compared to the polypeptide of SEQ ID NO: 1.