Polypeptides having nuclease activity

Patent metadata

Field	Value
Publication number	US-12221637-B2
Application number	US-201917276436-A
Country	US
Kind code	B2
Filing date	Oct 3, 2019
Priority date	Oct 4, 2018
Publication date	Feb 11, 2025
Grant date	Feb 11, 2025

Abstract

The present invention relates to polypeptides having nuclease activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and recombinant host cells comprising the polynucleotides as well as methods of producing, recovering and using the polypeptides. The invention further relates to cleaning compositions comprising one or more of the polypeptides having nuclease activity, as well as a cleaning method and use of the polypeptides.

First claim

The invention claimed is:

1. A cleaning composition comprising:
(a) at least 0.001 ppm of a polypeptide have nuclease activity, wherein the polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO: 96; and
(b) one or more surfactants.

2. The cleaning composition of claim 1 , wherein
the polypeptide comprises one or more of the motifs [HQ][FILVY]X[GAQS]DX[HTGSA][QVM]P[LFM]H (SEQ ID NO: 102), G[GA]NX[VILFY]X[VLM] (SEQ ID NO: 103), [SADN]R[GS]H (SEQ ID NO: 104), P[LM]H[VA][GA] (SEQ ID NO: 112), PLH[DN]E/(SEQ ID NO: 113), [YV][DN]RGH (SEQ ID NO: 114), YDRGHQ[AV] (SEQ ID NO: 115), [DNA]R[GSC]H[LI] (SEQ ID NO: 116) and/or [RIENLG][YF][RHN]V (SEQ ID NO: 117); and
the polypeptide belongs to the Pfam family PF02265 (S1-P1_nuclease), PF01223 (Endonuclease_NS) or PF13930 (Endonuclea_NS_2).

3. The cleaning composition of claim 2 , wherein the polypeptide is an S1-P1 nuclease that belongs to the Pfam family PF02265 (S1-P1_nuclease), and which comprises the motif [HQ][FILVY]X[GAQS]DX[HTGSA][QVM]P[LFM]H (SEQ ID NO: 102) and/or G[GA]NX[VILFY]X[VLM] (SEQ ID NO: 103).

4. The cleaning composition of claim 2 , wherein the polypeptide is of bacterial origin and comprises the motif P[LM]H[VA][GA] (SEQ ID NO: 112).

5. The cleaning composition of claim 2 , wherein the polypeptide is of fungal origin and comprises the motif PLH[DN]E (SEQ ID NO: 113).

6. The cleaning composition of claim 1 , wherein the polypeptide is an EN_NS nuclease that belongs to the Pfam family PF01223 (Endonuclease_NS) or PF13930 (Endonuclea_NS_2), and which comprises the motif [SADN]R[GS]H (SEQ ID NO: 104) and/or [YV][DN]RGH (SEQ ID NO: 114).

7. The cleaning composition of claim 6 , wherein the polypeptide is of fungal origin and comprises the motif YDRGHQ[AV] (SEQ ID NO: 115).

8. The cleaning composition of claim 6 , wherein the polypeptide is of bacterial origin and comprises the motif [DNA]R[GSC]H[LI] (SEQ ID NO: 116) and/or [RIENLG][YF][RHN]V (SEQ ID NO: 117).

9. The cleaning composition of claim 1 , wherein the polypeptide has at least 95% sequence identity to the polypeptide of SEQ ID NO: 96.

10. The cleaning composition of claim 1 , wherein the polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 96.

11. The cleaning composition of claim 1 , wherein the one or more surfactants are selected from anionic surfactants, nonionic surfactants, cationic surfactants, zwitterionic and mixtures thereof.

12. A method for laundering a textile, comprising the steps of:
(a) exposing the textile to a wash liquor comprising the cleaning composition of claim 1 ;
(b) completing at least one wash cycle; and
(c) optionally rinsing the textile.

13. An isolated polypeptide, which is
(a) a variant of the polypeptide of SEQ ID NO: 96, wherein the variant has nuclease activity and comprises one or more amino acid substitutions, and/or one or more amino acid deletions, and/or one or more amino acid insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 positions and wherein the variant has at least 90% sequence identity to the polypeptide of SEQ ID NO: 96; or
(b) a fragment of the polypeptide of SEQ ID NO:96, wherein the fragment has at least 90% sequence identity to the polypeptide of SEQ ID NO:96;

wherein the polypeptide has nuclease activity.

14. A recombinant host cell comprising a polynucleotide encoding the polypeptide of claim 13 operably linked to one or more control sequences that direct production of the polypeptide.

15. A method for producing a polypeptide having nuclease activity, comprising culturing the recombinant host cell of claim 14 under conditions for producing the polypeptide.

16. A recombinant host cell comprising a polynucleotide encoding a polypeptide having nuclease activity, wherein the polynucleotide is operably linked to one or more control sequences that direct production of the polypeptide and wherein the polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO: 96.

17. A method for producing a polypeptide having nuclease activity, comprising culturing the recombinant host cell of claim 16 under conditions for producing the polypeptide.

18. The method of claim 17 , wherein the polypeptide has at least 95% sequence identity to the polypeptide of SEQ ID NO: 96.

19. The polypeptide of claim 13 , wherein the variant has at least 95% sequence identity to the polypeptide of SEQ ID NO: 96.

20. The polypeptide of claim 13 , which is a fragment of the polypeptide of SEQ ID NO: 96.

Assignees

• Novozymes As

Inventors

• Sun tianqi
• Salomon Jesper
• Gori Klaus
• Gjermansen Morten
• Morant Marc Dominique
• Stringer Mary Ann

Classifications

• C11D2111/12

  Soft surfaces, e.g. textile · CPC title

• C11D3/38636

  containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase · CPC title

• C12N9/22 Primary

  Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title

• C11D3/386

  Preparations containing enzymes {, e.g. protease or amylase} · CPC title

• C12Y301/21001 Primary

  Deoxyribonuclease I (3.1.21.1) · CPC title