The present invention relates to a variant pullulanase, wherein the pullulanase comprises at least the following combination of substitutions: N368G+N393A+Q431E+L432F+A492A,S+N610R+G624S+T631S+S632C, and optionally further comprises N222P+Q252A+Q256R; wherein the variant has pullulanase activity, and wherein the variants have at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity to the polypeptide of SEQ ID NO: 3. Further aspect the present invention relates to a process for liquefying starch-containing material at a temperature above the initial gelatinization temperature using an alpha-amylase and a thermo-stable pullulanase of the invention.

The present disclosure provides methods and compositions for preparation of malted cereals, the improved malted cereals and their use, e.g., in the production of food and beverages.

A fungal alpha-amylase is provided from Aspergillus fumigatus (AfAmyl). AfAmyl has an optimal pH of 3.5 and is operable at 30-75 degrees C., allowing the enzyme to be used in combination with a glucoamylase and an isoamylase in a saccharification reaction. This obviates the necessity of running a saccharification reaction as a batch process, where the pH and temperature must be readjusted for optimal use of the alpha-amylase or glucoamylase. AfAmyl also catalyzes the saccharification of starch substrates to an oligosaccharide composition significantly enriched in DP2 and (DPI+DP2) compared to the products of saccharification catalyzed by an alpha-amylase from Aspergillus kawachii. This facilitates the utilization of the oligosaccharide composition by a fermenting organism in a simultaneous saccharification and fermentation process, for example.

The present disclosure relates to isolated polypeptides having alpha-amylase activity, polynucleotides encoding the polypeptides, nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing the polypeptides, and method of using polypeptides, including in ethanol production processes.

An object of the invention is to provide a beer-taste fermented malt beverage with an alcohol concentration of less than 1% (v/v), which is excellent in robust feeling of drinking, excellent in sweetness in harmony with sourness, and has beer-like flavor. The means for solving the problems is a beer-taste fermented malt beverage containing a wort fermented liquid, having a genuine extract component of 3.5% (w/w) or more, and an alcohol concentration of less than 1% (v/v).

Methods, compositions, and systems for steeping, propagation and fermentation, particularly large-scale operations for production of starch and ethanol and fermentation product streams are provided. Addition of mycotoxin mitigating enzymes or microorganisms expressing mycotoxin mitigating enzymes to steeping, propagation, and/or fermentation tanks, and/or to post-fermentation product streams, mitigates mycotoxin levels in fermentation co-products obtained from mycotoxin contaminated feedstocks.

A method of preparing a wort with an increased level of free amino nitrogen (FAN) comprising: a) preparing a mash from a grist comprising malt and/or adjunct; and b) adding a protease having at least 80% sequence identity to the polypeptide of SEQ ID NO: 1.

The present invention relates to the use of a cysteine endoprotease or a malt extract to prevent or reduce the cloudiness of a cereal-based beverage, fermented or not.

The present invention relates primarily to a method for obtaining a volatile fraction from a juice and/or an alcohol-containing beverage, comprising or consisting of the following steps: (a) distilling a mixture of substances from the juice and/or the alcohol-containing beverage, wherein the distillation is performed by a spinning cone column; (b) contacting the mixture of substances from step (a) with a sorbent to obtain a loaded sorbent; and (c) contacting the loaded sorbent from step (b) with a liquid desorbent to obtain the volatile fraction, wherein desired ingredients of the volatile fraction are concentrated as compared to the juice and/or alcohol-containing beverage such that an addition of 0.1% by mass or less of the volatile fraction to any food preparation produces a perceptible odor impression and/or taste impression in a test subject.

Methods, compositions, and systems for steeping, propagation and fermentation, particularly large-scale operations for production of starch and ethanol and fermentation product streams are provided. Addition of mycotoxin mitigating enzymes or microorganisms expressing mycotoxin mitigating enzymes to steeping, propagation, and/or fermentation tanks, and/or to post-fermentation product streams, mitigates mycotoxin levels in fermentation co-products obtained from mycotoxin contaminated feedstocks.