The present invention relates to a method of making glucose syrup from liquefied starch comprising, (a) contacting the liquefied starch with a glucoamylase, a pullulanase, and optionally an alpha-amylase wherein the ratio of pullulanase dose expressed as NPUN/gDS, to alpha-amylase dose expressed as FAU(A)/gDS is at least 60, particularly at least 75, particularly at least 100, more particularly at least 150, more particularly at least 200, more particularly at least 250, more particularly at least 300, more particularly at least 400, more particularly at least 500, more particularly at least 600, more particularly at least 800 or if no alpha-amylase is present the pullulanse is present in a dose of at least 0.5, particularly at least 0.75, particularly at least 1.0, particularly at least 1.5 NPUN/gDS, and (b) saccharifying the liquefied starch.

The present invention relates to processes of producing a fermentation product from starch containing material comprising (a) forming a slurry comprising the starch-containing material and water; (b) converting the starch-containing material into dextrins with an alpha-amylase; (c) saccharifying the dextrins using a carbohydrate source generating enzyme to form sugars; (d) fermenting sugars using a fermenting organism; (e) recovering the fermentation product to form whole stillage; (f) separating the whole stillage into a liquid fraction thin stillage and solid fraction wet cake; (g) hydrolyzing the thin stillage; (h) recycle a portion of the hydrolyzed thin stillage to steps (a); wherein the thin stillage in step (g) is hydrolyzed using a glucoamylase and/or polygalacturonase.

The present invention relates to a variant pullulanase, having increased thermo-stability and/or increased thermo-activity compared to a parent pullulanase, comprising a substitution at least a one position selected from a position corresponding to positions 432, 486, 370, 17, 77, 103, 106, 107, 190, 196, 197, 262, 279, 283, 321, 367, 375, 382, 399, 401, 402, 411, 412, 434, 435, 443, 446, 459, 460, 479, 490, 498, 514, 529, 531, 533, 541, 545, 581, 583, 595, 649, 665, 688, 700, 709, 804, 811 of SEQ ID NO: 1, and optionally a deletion of one or more, e.g., all amino acids at positions 821, 822, 823, 824, 825, 826, 827, and 828, wherein the variant has pullulanase activity, and wherein the variant has at least 70%, at least 75%, at least 80%, 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 a parent alpha amylase selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5. 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.

This specification discloses process for obtaining maltodextrin having DE between 17 and 19.9 and the maltodextrins obtained from the process. The disclosed maltodextrins can be provided as a powder or in shelf stable liquid form. The disclose maltodextrins have a polysaccharide profile similar to those observed for prior art maltodextrins, but make maltodextrin solutions having a high solids content, but reduced viscosity compared to prior art maltodextrins, on equivalent solids-in-solution basis. The process combines adds an alpha-amylase and a pullulanase enzyme to a polysaccharide mixture during a saccharification step. The disclosed maltodextrins make solutions at 50° C. and greater than 65% on a solids dry solids basis having a viscosity between 5,000 and 12,000 cP and having a water activity of less than 0.80.

The present invention aims to provide a saccharide composition suitable for a cyclic-tetrasaccharide-containing starch syrup which has low viscosity, low water activity, low coloration property, and low calorie content, and which is unlikely to cause precipitation of crystals of saccharides during storage, and to provide a use of the composition and a production method for the composition. The object is achieved by providing a cyclic-tetrasaccharide-containing saccharide composition having the following characteristics (1) to (3): (1) the saccharide composition includes a branched cyclic tetrasaccharide in addition to the cyclic tetrasaccharide, wherein the content of the cyclic tetrasaccharide with respect to the total solid content of the saccharide composition obtained by allowing glucoamylase and α-glucosidase to act on the above saccharide composition is 38% by mass or higher, on a dry solid basis; (2) the ratio of α-1,4-linked glucose in the total glucose residues constituting the saccharide composition in methylation analysis is over 9% and 15% or lower, and (3) the ratio of α-1,4,6-linked glucose in the total glucose residues constituting the saccharide composition in methylation analysis is less than 6%; and providing a use of the composition and a production method for the composition.

A process of recovering oil, comprising (a) converting a starch-containing material into dextrins with an alpha-amylase; (b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar; (c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism; (d) recovering the fermentation product to form a whole stillage; (e) separating the whole stillage into thin stillage and wet cake; (e′) optionally concentrating the thin stillage into syrup; (f) recovering oil from the thin stillage and/or optionally the syrup, wherein a phospholipase is present and/or added during steps (a) to (c). Use of phospholipase for increasing oil recovery yields from thin stillage and/or syrup in a fermentation product production process.

The present application relates to a psicose-6-phosphate phosphatase comprising motif A and motif B, a composition for producing D-psicose comprising the enzyme, and a method for producing D-psicose using the enzyme.

The present disclosure discloses a thermophilic recombinant type II pullulanase and the application thereof, and belongs to the technical field of genetic engineering. The present disclosure obtains a thermophilic recombinant type II pullulanase by heterologously expressing type II pullulanase in Escherichia coli. Its optimum pH is 6.6, it has better pH tolerance under the conditions of pH 5.8-8.0, and its optimum temperature is 95° C. After incubating at 95° C. for 10 h, the remaining enzyme activity is greater than 50%. It can exhibit higher specific enzyme activity under strong reducing conditions. For example, adding DTT to the culture environment can increase the specific enzyme activity of Sumo-Pul.sub.Py by 237.2%. The present disclosure also provides the combined truncation mutant Δ28N+Δ791C of type II pullulanase Sumo-Pul.sub.Py. The specific enzyme activity of the enzyme mutant is 32.18±0.92 U/mg, which is 5.99 times as high as that of the wild-type enzyme, thereby having important industrial application value and potential.

The present invention relates to processes for producing fermentation products from starch-containing material, wherein an alpha-amylase and a thermostable hemicellulase is present and/or added during liquefaction. The invention also relates to compositions suitable for use in processes of the invention.

A process of recovering oil, comprising (a) converting a starch-containing material into dextrins with an alpha-amylase; (b) saccharifying the dextrins using a carbohydrate source generating enzyme to form a sugar; (c) fermenting the sugar in a fermentation medium into a fermentation product using a fermenting organism; (d) recovering the fermentation product to form a whole stillage; (e) separating the whole stillage into thin stillage and wet cake; (e′) optionally concentrating the thin stillage into syrup; (f) recovering oil from the thin stillage and/or optionally the syrup, wherein a phospholipase is present and/or added during steps (a) to (c). Use of phospholipase for increasing oil recovery yields from thin stillage and/or syrup in a fermentation product production process.