Patent
The present invention is related to glucoamylases having at least 80% sequence identity to a Trichoderma glucoamylase having the sequence of SEQ ID NO: 4 and biologically functional fragments thereof. The invention is also related to DNA sequences coding for the glucoamylases, vectors and host cells incorporating the DNA sequences, enzyme compositions and methods of using the glucoamylases in various applications.
1. An isolated DNA sequence encoding an enzyme having glucoamylase activity, wherein the enzyme has at least 90% sequence identity to amino acid residues positions 1 to 452 of SEQ ID NO. 20.
2. The isolated DNA sequence of claim 1 , wherein the enzyme has an amino acid sequence that has at least 95% sequence identity to amino acid residues positions 1 to 452 of SEQ ID NO. 20.
3. The isolated DNA sequence of claim 1 , wherein the enzyme has an amino acid sequence that has at least 95% sequence identity to SEQ ID NO. 20.
4. The isolated DNA sequence of claim 1 , wherein the enzyme has an amino acid sequence that has at least 90% sequence identity to SEQ ID NO. 12.
5. A vector comprising the DNA of claim 1 .
6. An isolated host cell transformed with the vector of claim 5 .
7. A culture medium from the fermentation of a filamentous fungal host cell transformed with the DNA of claim 1 .
8. An isolated enzyme having glucoamylase activity, wherein the enzyme has at least 90% sequence identity to amino acid residues positions 1 to 452 of SEQ ID NO. 20.
9. The isolated enzyme of claim 8 , wherein the enzyme has at least 95% sequence identity to amino acid residues positions 1 to 452 of SEQ ID NO. 20.
10. The isolated enzyme of claim 8 , wherein the enzyme has an amino acid sequence that has at least 95% sequence identity to SEQ ID NO. 20.
11. The isolated enzyme of claim 8 , wherein the enzyme has an amino acid sequence that has at least 90% sequence identity to SEQ ID NO. 12.
12. A method of recombinantly producing a glucoamylase, comprising the step of expressing a polynucleotide encoding a polypeptide having glucoamylase activity in a filamentous fungal host cell, wherein the polypeptide comprises the amino acid sequence of claim 8 .
13. An enzyme composition comprising a glucoamylase having the amino acid sequence of claim 8 .
14. A method of hydrolyzing starch comprising treating a starch containing substrate with an enzyme having glucoamylase activity and at least 90% sequence identity to amino acid residues positions 1 to 452 of SEQ ID NO. 20 or a biologically active fragment thereof.
15. The method of claim 14 , wherein the starch is granular starch and the contacting step occurs at a temperature below the gelatinization temperature of the granular starch and at a pH of about 4 to 7.0 for a period of time to produce a composition comprising glucose.
16. The method of claim 15 , further comprising contacting the composition comprising glucose with a fermentation organism under suitable fermentation conditions to produce a fermentation product.
17. The method of claim 16 , wherein the fermentation product is ethanol.
18. The method of claim 14 , wherein the starch is liquefied starch.