Lipase variants and polynucleotides encoding same

• US 9,909,109 B2
• Filed: 03/22/2013
• Issued: 03/06/2018
• Est. Priority Date: 04/02/2012
• Status: Active Grant

• Current Assignee: Novozymes A/S (Novozymes)
• Original Assignee: Novozymes A/S (Novozymes)
• Inventors: Skjold-Joergensen, Jakob, Vind, Jesper, Svendsen, Allan
• Primary Examiner(s): Ekstrom, Richard C
• Application Number:
  US14/384,232
• Publication Number:
  US 20150031111A1
• Time in Patent Office: 1,810 Days
• Field of Search: None
• CPC Class Codes: C11D 3/38627 containing lipase
• CPC Class Codes: C12N 1/14 Fungi culture of mushrooms A01G18/00; as new plants A01H15/00; Culture media therefor
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 9/18 Carboxylic ester hydrolases (3.1.1)
• CPC Class Codes: C12N 9/20 Triglyceride splitting, e.g. by means of lipase

Abstract

The present invention relates to isolated lipase variants, comprising a substitution at one or more positions corresponding to positions I86D,E,N,Q, E87C, I90D,E,Q, and N92D,E,Q of the mature polypeptide of SEQ ID NO: 2, wherein the variant has lipase activity. In some embodiments the present invention relates to isolated polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; methods of producing the variants; and compositions comprising the variants. The present invention also relates to methods of obtaining lipase variants; methods of cleaning; and use of lipase variants for cleaning.

Claims

1. A lipase variant having at least 80% but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO:

2, and having lipase activity which consists or comprises any of the sets of substitutions corresponding to positions in the mature polypeptide of SEQ ID NO;

2;

a) S83T+E87T+G91 L+N92D+F95Y+D96T+L97P+K98Q+T231R+N233R;

b) I86P+E87A+G91A+N94D+D96I+L97F+K98D+T231R+N233R;

c) S83T+R84G+186D+E87Q+W89L+190Q+G91L+N92E+L93T+N94S+F95Y +D96T+K98T+T231R+N233R; and

d) D62C+S83T+E87C+G91L+N92D+F95Y+D96T+L97P+K98Q+T231R +N233R.

2. The variant of claim 1, which has an improved property relative to the parent, wherein the improved property is selected from:

improved catalytic rate;

improved specific activity;

improved substrate cleavage;

improved opening of the lid;

reduced calcium ion dependency;

improved builder tolerance, or improved wash performance.

3. An isolated polynucleotide encoding the variant of claim 1.

4. A nucleic acid construct comprising the polynucleotide of claim 3.

5. An expression vector comprising the polynucleotide of claim 3.

6. A host cell comprising the polynucleotide of claim 3.

7. A method of producing a lipase variant, comprising:

a) cultivating the host cell of claim 6 under conditions suitable for expression of the variant; and

b) recovering the variant.

8. A composition comprising the variant of claim 1.

9. A method of cleaning a textile or hard surface or other surface in fabric and home care comprising the steps of:

a) contacting the surface with an aqueous solution comprising the variant of claim 1; and

b) rinsing and drying the textile or hard surface.

10. The variant of claim 1, comprising the substitutions corresponding to positions S83T+R84G+I86D+E87Q+W89L+I90Q+G91L+N92E+L93T+N94S+F95Y+D96T+K98T+T231R+N233R of the mature polypeptide of SEQ ID NO:

2, wherein the variant has lipase activity.

11. The variant of claim 1, comprising the substitutions corresponding to positions D62C+S83T+E87C+G91L+N92D+F95Y+D96T+L97P+K98Q+T231R+N233R of the mature polypeptide of SEQ ID NO:

2, wherein the variant has lipase activity.

12. The variant of claim 1, comprising the substitutions corresponding to positions S83T+E87T+G91L+N92D+F95Y+D96T+L97P+K98Q+T231R+N233R of the mature polypeptide of SEQ ID NO:

2, wherein the variant has lipase activity.

13. The variant of claim 1, comprising the substitutions corresponding to positions 186P+E87A+G91A+N94D+D96I+L97F+K98D+T231R+N233R of the mature polypeptide of SEQ ID NO:

2, wherein the variant has lipase activity.