Polypeptides having xyloglucanase activity and nucleic acids encoding same
View Patent ↗The present invention relates to isolated polypeptides having Family 74 xyloglucanase activity and isolated nucleic acid sequences encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as methods for producing and using the polypeptides.
1. An isolated nucleic acid sequence encoding a polypeptide having Family 74 xyloglucanase activity comprising the amino acid sequence of SEQ ID NO: 2 or amino acids 20 to 838 of SEQ ID NO: 2.
2. The isolated nucleic acid sequence of claim 1 , encoding a polypeptide having Family 74 xyloglucanase activity consisting of the amino acid sequence of SEQ ID NO: 2 or amino acids 20 to 838 of SEQ ID NO: 2.
3. The isolated nucleic acid sequence of claim 1 produced by (a) hybridizing a population of DNA under high stringency conditions with (i) nucleotides 116 to 2740 of SEQ ID NO: 1, (ii) the cDNA sequence contained in nucleotides 116 to 2740 of SEQ ID NO: 1, or (iii) a complementary strand of (i) or (ii), wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/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 least at 65° C.; and (b) isolating the nucleic acid sequence from the population of DNA.
4. The isolated nucleic acid sequence of claim 1 , which is contained in plasmid pEJG81 which is contained in E. coli NRRL B-30502.
5. A nucleic acid construct comprising the nucleic acid sequence of claim 1 operably linked to one or more control sequences that direct the production of the polypeptide in a suitable expression host.
6. A recombinant expression vector comprising the nucleic acid construct of claim 5 .
7. An isolated host cell transformed with the nucleic acid construct of claim 5 .
8. A method for producing a polypeptide having Family 74 xyloglucanase activity comprising: (a) cultivating the host cell of claim 7 under conditions suitable for production of the polypeptide; and (b) recovering the polypeptide.
9. An isolated nucleic acid sequence comprising a nucleic acid sequence having at least one mutation in nucleotides 116 to 2740 of SEQ ID NO: 1, in which the mutant nucleic acid sequence encodes a polypeptide having Family 74 xyloglucanase activity comprising amino acids 20 to 838 of SEQ ID NO: 2.
10. A method for producing a polypeptide having Family 74 xyloglucanase activity comprising: (a) cultivating a host cell under conditions conducive for production of the polypeptide, wherein the host cell comprises the isolated nucleic acid sequence of claim 9 , and (b) recovering the polypeptide.