Polypeptide variants
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 polypeptide variant of a DNase parent polypeptide having the amino acid sequence SEQ ID NO: 1,
wherein the variant comprises one or both amino acid motifs [D/M/L][S/T]GYSR[D/N] (SEQ ID NO: 25) or ASXNRSKG (SEQ ID NO: 26),
wherein the variant comprises one or more amino acid substitution(s) compared to SEQ ID NO: 1,
wherein the substitution is selected from the group consisting of: S13D, S13Y, S13Q, S13F, S13R, S13V, S13N, S13H, S13M, S13W, S13K, S13L, S13E, T22P, T22A, T22V, T22D, T22R, T22K, T22M, T22E, T22H, T22L, T22W, T22F, T22C, T22I, S39A, S39P, S39Y, S39H, S39E, S39N, S39M, S39D,
wherein the variant has an amino acid sequence identity to the parent polypeptide of at least 90%,
and the variant has DNase activity.
2. The polypeptide variant of claim 1 , further comprising one or more substitutions selected from the group consisting of: T1I, T1V, T1Y, T1M, T1E, G4N, T5F, T5C, P6V, P6G, S7D, S7T, K8V, S9K, S9Q, S9V, S9L, S9F, S9P, S9R, A10D, A10M, A10I, A10K, Q12S, Q12V, Q12E, Q14M, Q14R, N16S, A17C, A17V, A17E, A17T, T19K, T19L, T19S, T19I, T19V, K21E, K21M, G24Y, S25P, S27N, S27I, S27M, S27D, S27V, S27F, S27A, S27C, S27L, S27E, G28L, Y29W, S30K, S30D, S30H, S30T, D32Q, I38V, I38M, Q40V, S42C, S42L, S42M, S42F, S42W, V49R, L51I, K52I, K52H, A55S, D56I, D56L, D56T, S57W, S57F, S57H, S57C, S57P, S57V, S57R, S57T, Y58A, Y58T, S59C, S59T, S59L, S59Q, S59V, S59K, S59R, S59M, S59I, S59H, N61 D, P63A, T65L, T65I, T65V, T65R, T65K, S68V, S68I, S68W, S68Y, S68H, S68C, S68T, S68L, V76G, V76L, V76C, V76K, V76H, V76E, V76A, V76Y, V76N, V76M, V76R, V76F, T77N, T77Y, T77W, T77R, F78I, F78H, F78Y, F78C, T79G, T79R, N80K, S82L, S82E, S82K, S82R, S82H, D83C, D83F, D83L, L92T, A93G, E94N, G99S, S101 D, S101A, S102M, S102L, S102V, S102A, S102K, S102T, S102R, T104P, T104A, T105V, T105I, K107L, K107C, K107R, K107H, K107S, K107M, K107E, K107A, K107D, Q109R, Q109S, A112S, S116D, S116R, S116Q, S116H, S116V, S116A, S116E, S116K, A125K, S126I, S126E, S126A, S126C, T127C, T127V, S130E, G132R, D135R, T138Q, W139R, R143E, R143K, S144Q, S144H, S144L, S144P, S144E, S144K, G145V, G145E, G145D, G145A, A147Q, A147W, A147S, G149S, K152H, K152R, S156C, S156G, S156K, S156R, S156T, S156A, T157S, Y159F, K160V, W161L, W161Y, G162Q, G162D, G162M, G162R, G162A, G162S, G162E, G162L, G162K, G162V, G162H, S164R, S164T, Q166D, S167M, S167L, S167F, S167W, S167E, S167A, S167Y, S167H, S167C, S167I, S167Q, S167V, S167T, S168V, S168E, S168D, S168L, K170S, K170L, K170F, K170R, T171 D, T171E, T171A, T171C, A172G, A172S, L173T, L173A, L173V, Q174L, G175D, G175E, G175N, G175R, M176H, L177I, N178D, N178E, N178T, N178S, N178A, S179E, S181R, S181E, S181D, S181F, S181H, S181W, S181L, S181M, S181Y, S181Q, S181G, S181A, Y182M, Y182C, Y182K, Y182G, Y182A, Y182S, Y182V, Y182D, Y182Q, Y182F, Y182L, Y182N, Y182I, Y182E, Y182T and Y182W compared to SEQ ID NO: 1.
3. The polypeptide variant of claim 1 , which has an improved stability, measured as half-life improvement factor, HIF, compared to the parent polypeptide.
4. The polypeptide variant of claim 1 , wherein the variant has at least 95% but less than 100% sequence identity to the parent polypeptide.
5. The polypeptide variant of claim 1 , wherein the variant has at least 96% but less than 100% sequence identity to the parent polypeptide.
6. The polypeptide variant of claim 1 , wherein the variant has at least 97% but less than 100% sequence identity to the parent polypeptide.
7. The polypeptide variant of claim 1 , wherein the variant has at least 98% but less than 100% sequence identity to the parent polypeptide.
8. The polypeptide variant of claim 1 , wherein the variant has at least 99% but less than 100% sequence identity to the parent polypeptide.
9. The polypeptide variant of claim 1 , wherein the total number of amino acid substitutions compared to SEQ ID NO: 1 is 1-20.
10. A detergent composition, comprising the polypeptide variant of claim 1 .
11. The detergent composition of claim 10 , further comprising one or more detergent components.
12. The detergent composition of claim 10 , further comprising one or more additional enzymes selected from the group consisting of proteases, amylases, lipases, cutinases, cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases, peroxidaes, haloperoxygenases, catalases, mannanases, and any mixture thereof.
13. The detergent composition of claim 10 , in the 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.
14. A method, comprising,
exposing a textile or hard surface to the detergent composition of claim 10 , and
optionally rinsing the textile or hard surface.
15. A polynucleotide encoding the variant of claim 1 .
16. A nucleic acid construct comprising the polynucleotide of claim 15 .
17. An expression vector comprising the polynucleotide of claim 15 .
18. A recombinant host cell comprising the polynucleotide of claim 15 .
19. A method of producing a DNase variant, comprising: cultivating the recombinant host cell of claim 18 under conditions suitable for expression of the variant; and optionally recovering the variant.