Processes for producing a fermentation product
The present invention relates to polypeptides having trehalase activity, particularly derived from Talaromyces . 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 for the production of ethanol.
1. A process of producing a fermentation product, comprising (a) liquefying a starch-containing material with an alpha-amylase;
optionally presaccharifying the liquefied material before step (b);
(b) saccharifying the liquefied material;
(c) fermenting using a fermentation microorganism;
wherein
i) a glucoamylase; and
ii) a polypeptide having trehalase activity selected from the group consisting of a polypeptide having at least 93% sequence identity to the mature polypeptide of SEQ ID NO: 21 and a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 20 or the cDNA sequence thereof; and
iii) optionally a cellulolytic enzyme composition, a protease, or both a cellulolytic enzyme composition and a protease are present or added during
the saccharification step (b);
the fermentation step (c); or
the optionally presaccharification step before step (b).
2. The process of claim 1 , wherein an alpha-amylase is present and/or added during saccharification step (b); fermentation step (c); or the optional presaccharification step before step (b).
3. The process of claim 1 , wherein the alpha-amylase is the one disclosed as SEQ ID NO: 7, having one or more of the following substitutions: G128D, D143N, or G128D+D143N (using SEQ ID NO: 7 for numbering), and wherein the alpha-amylase variant has at least 80%, but less than 100% identity to the mature part of the polypeptide of SEQ ID NO: 7.
4. The process of claim 1 , wherein the fermenting microorganism is derived from a strain of Saccharomyces.
5. The process according to claim 4 , wherein the yeast fermenting organism expresses the polypeptide having trehalase activity.
6. A process of producing fermentation products from starch-containing material comprising:
(i) saccharifying a starch-containing material at a temperature below the initial gelatinization temperature; and
(ii) fermenting using a fermentation microorganism;
wherein saccharification, fermentation, or saccharification and fermentation is done in the presence of the following enzymes:
a. glucoamylase;
b. alpha-amylase; and
c. a polypeptide having trehalase activity selected from the group consisting of a polypeptide having at least 93% sequence identity to the mature polypeptide of SEQ ID NO: 21 and a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 20 or the cDNA sequence thereof; and
d. optionally a protease, a cellulolytic enzyme composition, or a protease and a cellulolytic composition.
7. The process of claim 1 , wherein steps (b) and (c) are performed simultaneously in a simultaneous saccharification and fermentation.
8. The process of claim 7 , wherein an alpha-amylase is present or added during the simultaneous saccharification and fermentation.
9. The process of claim 7 , wherein the cellulolytic enzyme composition is present or added during the simultaneous saccharification and fermentation.
10. The process of claim 7 , wherein the protease is present or added during the simultaneous saccharification and fermentation.