The present invention relates to polynucleotides encoding novel fusion polypeptides essentially composed of a signal peptide for membrane translocation and a polypeptide providing -1,6-glucosidase activity and to bacteria containing said polynucleotides. The invention further relates to methods for producing fine chemicals using media containing isomaltose and/or panose as carbon source.

The present invention relates to processes for producing fermentation products from starch-containing material, wherein an alpha-amylase and optionally a thermostable protease, pullulanase and/or glucoamylase are present and/or added during liquefaction, wherein a cellulolytic composition is present and/or added during fermentation or simultaneous saccharification and fermentation. The invention also relates to a composition suitable for use in a process of the invention.

The present invention relates to pullulanase variants comprising substitutions of the parent pullulanase at one or more positions corresponding to positions 393, 143, 150, 243, 244, 345, 346, 368, 370, 373, 381, 382, 385, 387, 402, 429, 430, 431, 432, 456, 486, 492, 610, 624, 631, 632, 665 and 699 of the polypeptide of SEQ ID NO: 3. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

In one aspect, the invention is directed to polypeptides having an amylase activity, polynucleotides encoding the polypeptides, and methods for making and using these polynucleotides and polypeptides. The polypeptides of the invention can be used as amylases to catalyze the hydrolysis of starch into sugars.

An enzymatically produced soluble ?-glucan fiber composition is provided suitable for use as a digestion resistant fiber in food and feed applications. The soluble ?-glucan fiber composition can be blended with one or more additional food ingredients to produce fiber-containing compositions. Methods for the production and use of compositions comprising the soluble ?-glucan fiber are also provided.

The present invention relates to processes for producing fermentation products from starch-containing material, wherein an alpha-amylase and optionally a thermostable protease, pullulanase and/or glucoamylase are present and/or added during liquefaction, wherein a cellulolytic composition is present and/or added during fermentation or simultaneous saccharification and fermentation. The invention also relates to a composition suitable for use in a process of the invention.

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 conventional corn dry grind ethanol production process requires the addition of exogenous alpha and glucoamylase enzymes to break down starch into glucose, which is fermented to ethanol by yeast. The present invention describes use of new genetically engineered corn and yeast, which can eliminate or minimize the use of these external enzymes, improve the economics and process efficiencies, and simplify the process.

Provided herein are double strand DNA molecules comprising inverted repeats, expression cassette and one or more restriction sites for nicking endonucleases, the methods of use thereof, and the methods of making therefor.

A method for making a mutated pullulanase enzyme that hydrolyzes an -1,6-glycosidic linkage is provided. The method includes obtaining the amino acid sequence of a pullulanase enzyme having an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 13; identifying an amino acid to be mutated in the pullulanase enzyme of step (1), wherein the amino acid to be mutated corresponds to the amino acid at position Phe476 of SEQ ID NO: 2; constructing a mutated amino acid sequence by substituting the amino acid to be mutated with another amino acid or deleting the amino acid to be mutated, thereby making a mutated pullulanase enzyme having the mutated amino acid sequence that has increased affinity for pullulan and that hydrolyzes an -1,6-glycosidic linkage.