Dnase variants

Abstract

The present invention relates to polypeptide variants and methods for obtaining variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

1 . A DNase variant, comprising an substitution at one or more positions corresponding to positions 4, 17, 19, 36, 38, 40, 41, 45, 51, 53, 54, 55, 57, 64, 66, 67, 68, 69, 70, 71, 72, 74, 75, 77, 82, 83, 84, 85, 86, 88, 91, 99, 101, 105, 105, 106, 115, 116, 135, 136, 138, 139, 140, 141, 151, 152, 153, 154, 162, 163, 164, 166, 168, 169, 173, 182, 183, 184, 185, 186, 189, 189, 212, and 215 of SEQ ID NO: 1, wherein the variant has a sequence identity to the polypeptide shown in SEQ ID NO: 1 of at least 80%, wherein the variant has DNase activity and wherein the variant has improved stability as compared to the SEQ ID NO 1. 
     
     
         2 . The DNase variant of  claim 1 , wherein the variant comprises one or more substitutions selected from the group consisting of N4E, L17E, T19A, T19G, T19I, K36P, Q38P, A40P, A40H, L41T, L41H, V45H, L51G, K53T, K53P, G54P, A55P, N57H, E64A, E64Q, E64R, E64T, E64I, E64S, T66H, K67A, K67T, N68V, N68P, N68I, N68H, S69A, S69D, S69E, S69K, S69L, S69W, S69Y, S69Q, N70T, N70H, N70G, R71T, D72E, S74H, S74G, G75I, N77T, K82P, K82I, D83T, D83P, D83I, D83H, D83G, P84H, Q85T, Q85P, Q85H, K86T, K86P, K86H, G88P, G88H, A91P, W99T, A101W, K105E, K105N, K105T, K105D, S106T, S115T, L116I, Q135L, G136L, V138I, V138L, V138P, V138Q, L139A, N140R, N140L, N140A, G141L, F151R, D152Y, D152L, D152I, D152A, P153E, S154R, T162R, W163E, F164R, I166Y, I166R, K168N, F169R, F169E, A173I, A173R, A173T, S182R, N183E, D184I, K185Y, S186I, D189G, D189H, K212G, K212P and K215I, wherein the positions correspond to the positions of SEQ ID NO 1. 
     
     
         3 . The DNase variant according to  claim 1 , wherein the DNase variant has an Improvement Factor (IF) of at least 1.1; 1.5; 2.0; 3.0, 3.5, 4.0, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10 compared to the polypeptide shown in SEQ ID NO: 1. 
     
     
         4 . The variant of  claim 1 , wherein the variant has at least 90%, sequence identity to the mature polypeptide of SEQ ID NO: 1. 
     
     
         5 . The variant of  claim 1 , wherein the total number of alterations compared to SEQ ID NO: 1 is 1-20. 
     
     
         6 . A composition comprising a DNase variant according to  claim 1 , wherein the composition comprises:
 a) at least 0.002 ppm DNase variant, optionally   b) 2 wt % to 60 wt % of at least one surfactant, optionally   c) 5 wt % to 50 wt % of at least one builder, and/or optionally at least one additional enzyme.   
     
     
         7 . The composition of  claim 6 , wherein the composition is a detergent composition. 
     
     
         8 . The composition according to  claim 6 , wherein the composition further comprising one or more additional enzymes selected from the group comprising of proteases, amylases, lipases, cutinases, cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases, peroxidaes, haloperoxygenases, catalases and mannanases, or any mixture thereof. 
     
     
         9 . The composition according to  claim 6  in form of a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid. 
     
     
         10 . (canceled) 
     
     
         11 . A method for obtaining a DNase variant, comprising
 (a) introducing into a parent DNase an alteration at one or more positions corresponding to positions 4, 17, 19, 36, 38, 40, 41, 45, 51, 53, 54, 55, 57, 64, 66, 67, 68, 69, 70, 71, 72, 74, 75, 77, 82, 83, 84, 85, 86, 88, 91, 99, 101, 105, 105, 106, 115, 116, 135, 136, 138, 139, 140, 141, 151, 152, 153, 154, 162, 163, 164, 166, 168, 169, 173, 182, 183, 184, 185, 186, 189, 189, 212, or 215 of SEQ   (b) ID NO 1, wherein the variant has an amino acid sequence which is at least 80% identical to SEQ ID NO 1, and   (c) recovering the variant.   
     
     
         12 . The method of  claim 11 , wherein the variant comprises two, three, four or five alterations compared to SEQ ID NO: 1. 
     
     
         13 . The method according to  claim 11 , wherein the DNase variant comprises one or more substitution selected from the group consisting of N4E, L17E, T19A, T19G, T19I, K36P, Q38P, A40P, A40H, L41T, L41H, V45H, L51G, K53T, K53P, G54P, A55P, N57H, E64A, E64Q, E64R, E64T, E64I, E64S, T66H, K67A, K67T, N68V, N68P, N68I, N68H, S69A, S69D, S69E, S69K, S69L, S69W, S69Y, S69Q, N70T, N70H, N70G, R71T, D72E, S74H, S74G, G75I, N77T, K82P, K82I, D83T, D83P, D83I, D83H, D83G, P84H, Q85T, Q85P, Q85H, K86T, K86P, K86H, G88P, G88H, A91P, W99T, A101W, K105E, K105N, K105T, K105D, S106T, S115T, L116I, Q135L, G136L, V138I, V138L, V138P, V138Q, L139A, N140R, N140L, N140A, G141L, F151R, D152Y, D152L, D152I, D152A, P153E, S154R, T162R, W163E, F164R, I166Y, I166R, K168N, F169R, F169E, A173I, A173R, A173T, S182R, N183E, D184I, K185Y, S186I, D189G, D189H, K212G, K212P and K215I, compared to SEQ ID NO: 1. 
     
     
         14 . The method according to  claim 11 , wherein the alteration introduced into the parent DNase is a deletion of the amino acid in one or more of the positions 84, 88, 139, 179 or 180. 
     
     
         15 . The method according to  claim 11 , wherein the parent DNase is the polypeptide comprising the amino acid sequence shown in SEQ ID NO 1 or a polypeptide having at least 80% sequence identity hereto. 
     
     
         16 . The method according to  claim 11 , wherein the DNase variant has at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide shown in SEQ ID NO: 1.