A brewing method
A technology of amylase and auxiliary materials, which is applied in beer brewing, biochemical equipment and methods, enzymes, etc., can solve the problems of increasing the proportion of auxiliary materials, unwillingness and unwillingness of brewers to use them.
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Embodiment 1
[0114] The purpose of this example was to demonstrate the benefit of the presence of all 3 enzymes (pullulanase, maltogenic alpha-amylase and alpha-amylase) over the presence of only two of them in different combinations and dosages during mashing.
[0115] Maize grist comprising 50% unpregelatinized maize grist (maizegrist) and 50% fully modified (WM; well modified) malt was ground (0.2 mm gap (gap) disc mill) and prepared in Seq ID No. .3 starch saccharification is carried out in the presence of the pullulanase of 3, the α-amylase of Seq ID No.2 and the maltogenic α-amylase of Seq ID No.1. Corn and malt starches were mashing-in at a ratio of 1:5 (normal gravity), and the mashing-in temperature profile consisted of mashing-in at 52°C for 30 minutes, increasing to 62°C (1°C / min), Hold at 62°C for 30 minutes, increase to 72°C (1°C / min), hold at 72°C for 30 minutes, increase to 78°C (1°C / min), then immediately cool to 20°C (total saccharification time 131 minutes ). Wort and f...
Embodiment 2
[0121] The objective was to evaluate the temperature optimum of β-amylase from Clostridium thermosulfogenes.
[0122] The temperature optimum of the beta-amylase of Seq ID No. 4 obtained from Clostridium thermosulfogenes was measured by determining the release of maltose from amylopectin by measuring the reducing end using PHABH reagent. 1 mL of substrate (1% w / v potato pullulan in 50 mM NaOAc, 1 mM calcium chloride, pH 4.5) was pre-incubated at 60°C for 10 minutes. As a control, 150 uL was removed and mixed with 75 uL of stop reagent (0.75 g PHABH (Sigma), 2.5 g potassium sodium tartrate (Merck) and 50 ml 2% NaOH) before adding the enzyme. For enzyme incubation, 150 uL of enzyme solution was added to the substrate and incubated at 30, 40, 50, 60, 70, or 80°C for 10 minutes. 75 μL of stop reagent was added and the solution was incubated at 100° C. for 15 minutes. Transfer 200 μL to a PCR tube and measure absorbance at 410 nm. The experiments were performed in triplicate. T...
Embodiment 3
[0129] The purpose of this example was to demonstrate the benefit of the presence of a thermostable beta-amylase in combination with pullulanase and alpha-amylase during saccharification and filtration.
[0130] Grits containing 50% unpregelatinized corn meal and 50% fully modified (WM) malt were ground (disc mill with 0.2mm gap) and prepared in the pullulanase of Seq ID No.3, Seq Starch saccharification was carried out in the presence of α-amylase of ID No.2 and β-amylase of Seq ID No.4. Saccharify corn and malt starch at a ratio of 1:5 (normal gravity), and the composition of the starch saccharification temperature distribution is: saccharify starch at 52°C for 30 minutes, increase to 62°C (1°C / min), and maintain at 62°C for 30 minutes , increase to 72°C (1°C / min), hold at 72°C for 30 minutes, increase to 78°C (1°C / min) and incubate at 78°C for 2 hours, immediately after cooling to 20°C. Wort and fermented wort (one day laboratory fermentation at room temperature) were anal...
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