Recombinant production of glucoamylase P in trichoderma
AssigneeAlko-Yhiot Oy
InventorsTuula Torkkeli, Vesa Joutsjoki, Helena Torkkeli, Arja Vainio, Richard Fagerstrom, Sirpa Aho, Matti Korhola, Helena Nevalainen
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Abstract
The invention is directed to amino acid and DNA sequences of a unique glucoamylase P that has a high debranching activity, a Trichoderma host cell, transformed with such sequences, the expression of such recombinant glucoamylase P, and the industrial uses for the recombinant enzyme and hosts transformed therewith.
Claims35
A composition comprising purified DNA molecules that have a nucleotide sequence capable of being processed by a Trichoderma host cell to encode a protein, said protein comprising an amino acid sequence selected from the group consisting of:
- a. amino acids 1-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- and
- b. amino acids 30-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- said DNA molecules also having the sequence of introns IVS1, IVS2 and IVS3 as shown in SEQ ID NO. 4 (FIG. 13), said introns being inserted between exons encoding said amino acids as shown in SEQ ID NO. 4 (FIG. 13).
A recombinant vector comprising a DNA sequence that is capable of being processed by a Trichoderma host cell to encode a protein, said protein comprising an amino acid sequence selected from the group consisting of:
- a. amino acids 1-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- and
- b. amino acids 30-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- said DNA sequence also having the sequence of introns IVS1, IVS2 and lVS3 as shown in SEQ ID NO. 4 (FIG. 13), said introns being inserted between exons encoding said amino acids as shown in SEQ ID. NO, 4 (FIG. 13 ).
A Trichoderma host cell transformed with a DNA molecule having a nucleotide sequence that is capable of being processed by said Trichoderma host cell to encode a protein, said protein comprising an amino acid sequence selected from the group consisting of:
- a. amino acids 1-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- and
- b. amino acids 30-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- said DNA molecule also having the sequence of introns IVS1, IVS2 and IVS3 as shown in SEQ ID NO. 4 (FIG. 13), said introns being inserted between exons encoding said amino acids as shown in SEQ ID NO. 4 (FIG. 13).
A Trichoderma host cell transformed with DNA encoding amino acids 1-29 of SEQ ID NO. 1 (FIG. 5) or in SEQ ID NO. 4 (FIG. 13).
The culture medium from the fermentation of a Trichoderma host cell that is transformed with a DNA molecule having a molecule sequence capable of being processed by said Trichoderma host cell to encode a protein, said protein comprising an amino acid sequence selected from the group consisting of:
- a. amino acids 1-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- and
- b. amino acids 30-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- said DNA molecule also having the sequence of introns IVS1, IVS2 and IVS3 as shown in SEQ ID NO. 4 (FIG. 13), said introns being inserted between exons encoding said amino acids as shown in SEQ ID NO. 4 (FIG. 13).
A method for producing glucoamylase P, wherein said method comprises expression of said glucoamylase P from Trichoderma, wherein said DNA encoding said glucoamylase P has a DNA sequence encoding, or capable of being processed by said Trichoderma host cell to encode, a protein, said protein comprising an amino acid sequence selected from the group consisting of:
- a. amino acids 1-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13);
- and
- b. amino acids 30-616 as shown in SEQ ID NO. 2 (FIG. 5A) or in SEQ ID NO. 4 (FIG. 13).
Dependencies are inferred from the claim text.