Polynucleotides encoding polypeptides having phospholipase C activity

Patent metadata

Field	Value
Publication number	US-9279110-B2
Application number	US-201113884063-A
Country	US
Kind code	B2
Filing date	Nov 9, 2011
Priority date	Nov 12, 2010
Publication date	Mar 8, 2016
Grant date	Mar 8, 2016

Abstract

The present invention relates to isolated polypeptides having phospholipase C activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

First claim

The invention claimed is:

1. A nucleic acid construct comprising a polynucleotide encoding a polypeptide having phospholipase C activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct expression of the polypeptide in an expression host cell, and wherein the amino acid sequence of the polypeptide having phospholipase C activity is selected from the group consisting of:
(a) an amino acid sequence with at least 90% sequence identity to the sequence of amino acids 1 to 625 of SEQ ID NO: 2 and
(b) a fragment of the sequence of amino acids 1 to 625 of SEQ ID NO: 2 that has phospholipase C activity.

2. A microbial recombinant host cell comprising the nucleic acid construct of claim 1 .

3. A method of producing a polypeptide having phospholipase C activity, comprising:
(a) cultivating the microbial recombinant host cell of claim 2 under conditions conducive for production of the polypeptide having phospholipase C activity; and
(b) recovering the polypeptide.

4. The nucleic acid construct of claim 1 , wherein the polypeptide having phospholipase C activity has an amino acid sequence with at least 95% sequence identity to the sequence of amino acids 1 to 625 of SEQ ID NO: 2.

5. The nucleic acid construct of claim 1 , wherein the polypeptide having phospholipase C activity has an amino acid sequence with at least 97% sequence identity to the sequence of amino acids 1 to 625 of SEQ ID NO: 2.

6. The nucleic acid construct of claim 1 , wherein the polypeptide having phospholipase C activity comprises the sequence of amino acids 1 to 625 of SEQ ID NO: 2.

7. The nucleic acid construct of claim 1 , wherein the polypeptide having phospholipase C activity is encoded by a polynucleotide that hybridizes with the full complementary sequence of nucleotides 55 to 1929 of SEQ ID NO: 1 under very high stringency conditions, wherein the very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.

Assignees

• Tang Lan
• Liu Ye
• Borch Kim
• Brask Jesper
• Novozymes As

Inventors

• Tang Lan
• Liu Ye
• Borch Kim
• Brask Jesper

Classifications

• C11B3/003 Primary

  by enzymes or microorganisms, living or dead · CPC title

• C12N9/20 Primary

  Triglyceride splitting, e.g. by means of lipase · CPC title

• C12Y301/04003

  Phospholipase C (3.1.4.3) · CPC title

• C12N9/16

  acting on ester bonds (3.1) · CPC title

• A23D9/04 Primary

  Working-up · CPC title