Enzymes for infusion mashing in adjunct brewing technical field
A technology for assisting brewing and glucoamylase, applied in the directions of glycosylase, beer brewing, enzymes, etc.
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[0117] Example 1 - Enzyme
[0118] GsAA1: an alpha-amylase variant from Geobacillus stearothermophilus, having the amino acid sequence shown in SEQ ID NO:1.
[0119] GsAA2: maltogenic alpha-amylase from Geobacillus stearothermophilus, having the amino acid sequence shown in SEQ ID NO:2.
[0120] TrGA: glucoamylase from Trichoderma reesei, having the amino acid sequence shown in SEQ ID NO:3.
[0121] As an example of an alpha-amylase, used from DuPont (DuPont) 5T (A 5T). As an example of a maltogenic alpha-amylase from DuPont was used MA (D MA). As an example of glucoamylase from DuPont is used TGA (D TGA).
[0122] Example 2 - Protein Assay Method
[0123] Protein Assay by Stain Free Imager Criterion
[0124] Using Gel Doc TM The EZ Imaging System quantifies proteins by SDS-PAGE gels and densitometry. Reagents used in the assay: Concentrated (2x) Laemmli sample buffer (Bio-Rad, Cat. No. 161-0737); 26-well XT4-12% Bis-Tris gel (Bio-Rad , catalog number 345-0125)...
example 3
[0125] Example 3 - Sugar Analysis by HPLC
[0126] All standards: Glucose, maltose, maltotriose and maltotetraose were prepared in double distilled water (ddH20) and filtered through a 0.45 μm syringe filter. A series of each standard was prepared at concentrations ranging from 10 to 100,000 ppm.
[0127] All wort samples containing active enzymes were inactivated by heating the samples to 95°C for 10 min. Wort samples were subsequently prepared in 96-well MTP plates (Corning, NY, USA) and diluted at least 4 times in ddH2O and filtered through a 0.20 μm 96-well plate filter (Corning filter plate , PVDF hydrophilic membrane, New York State, USA). All samples were analyzed in duplicate.
[0128] instrument
[0129] Quantification of sugars: DP1, DP2, DP3, DP4 and DP5+ by UPLC. Sample analysis was carried out in a laboratory equipped with DGP-3600SD dual-gradient analysis pump, WPS-3000TSL constant temperature autosampler, TCC-3000SD constant temperature column oven and RI...
example 4
[0132] Example 4 - Low temperature leach saccharification of corn, rice, sorghum and cassava using enzymes to obtain extracts and fermentable sugars
[0133] The purpose of this example is to demonstrate the presence of two enzymes (maltogenic alpha amylase or glucoamylase and alpha amylase ) benefits (fermentable sugars and extract release).
[0134] The enzymes were tested in a model system of mashing operations for wort production using corn grain (Nordgetreide GmBH Lübec, Germany), rice grain (Cambodia, MEKONG Asian Market), Dagrofa Brabrand), sorghum (white unground sorghum - Diageo, Ireland) and tapioca (Uganda) with a fixed water to grain ratio of 4:1. Rice grains were ground on a Buhler Miag malt mill at a setting of 0.5 mm and sorghum at a setting of 1.6 mm.
[0135] Mashing operations for wort production
[0136] Maize grains (3.0 g), rice grains (ground 3.0 g), sorghum (3.0 g) or tapioca flour (3.0 g) were mixed in wheaton cups (glass containers with lids) and ...
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