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An amylase and high-activity technology, applied in the direction of hydrolytic enzymes, etc., can solve the problems of limited application range, poor temperature resistance, and low β-amylase activity, and achieve high sugar production efficiency enhancement, excellent product characteristics, and COD reduction. Effect
Active Publication Date: 2015-03-04
山东绿健生物技术有限公司
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Problems solved by technology
[0004] However, domestic production of β-amylase from sweet potato and wheat has low activity (500,000 u / ml), poor temperature resistance (58°C), and does not meet the production requirements of high maltose and ultra-high maltose syrup.
Although the imported high-activity β-amylase can meet the production requirements of high-maltose syrup, because its temperature resistance is not as good as that of soybean β-amylase, the amount of β-amylase used in the production of high maltose is large, and the production of high maltose The necessary amount of pullulanase is also greater than that of soybean β-amylase (caused by the narrow pH of barley β-amylase and the inconsistent pH of pullulanase)
[0005] The process of the above-mentioned high-activity soybean β-amylase is applied to industrial production, but the scope of use is limited, the price is high, and the demand cannot be met
[0006] At the same time, although domestic scientific research departments have obtained many scientific research achievements in the preparation of β-amylase, most of them have high energy consumption, high environmental pollution (salting out method), low comprehensive utilization rate of raw materials, and high cost (raw materials used Low-temperature soybean meal, the price is more than 3,500 yuan / ton) (see the Chinese invention patent titled "Preparation Process of Soybean β-amylase" and patent number 99102506)
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Embodiment 1 preparation example 1
[0048] Take 100L of separated water (wastewater after protein separation) (activity 4125u / ml), adjust the temperature to 45°C, add 0.1% PAA to pH 3.4 to obtain a precipitate, add 100g of calcium carbonate after dissolution, neutralize the pH to 6.30, and pass through 300cm 2 Filtrate through a filter, concentrate the filtrate with a mesoporous fiber membrane, add an enzyme stabilizer (salt, dextrin, sodium acetate), and refine to obtain 416ml of a finished product of 716,400 u / ml.
Embodiment 2 preparation example 2
[0050] Take 1500L of collected and separated water (activity 4380u / ml), adjust the temperature to 44°C, add 0.2% PAA to pH 3.3 to obtain a precipitate, add water 1:5 to dissolve, add 1.1kg of calcium carbonate to neutralize pH 6.05, and use 1m 2 The filtrate was obtained by plate and frame filtration, concentrated by 4-inch roll-type membrane ultrafiltration, and added with enzyme stabilizers (salt, dextrin, sodium acetate), refined to obtain 6589ml of 745,800 u / ml finished product.
Embodiment 3 preparation example 3
[0052] Take and collect separated water 10M 3 (Activity 4217u / ml) Adjust the temperature to 44°C, add 0.50% PAA to obtain a precipitate, add 1:5 water to dissolve, add 10kg calcium carbonate to neutralize pH 6.18, and use 8-inch roll membrane ultrafiltration Concentrate and refine with enzyme stabilizer (salt, dextrin, sodium acetate) to obtain 34787ml of 916,400 u / ml finished product.
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Abstract
The invention relates to a separation and purification method for amylase in a plant, and particularly relates to a method for industrially producing high-activity beta-amylase. The preparation method for the beta-amylase disclosed by the invention comprises the following steps of: collecting separation water; pre-treating; precipitating and dissolving out the beta-amylase; forming a composite precipitate from polyacrylic acid and the beta-amylase in the solution; dissolving the composite precipitate; and adjusting the pH to be greater than or equal to 6.0, and dissolving out the beta-amylase. The invention provides a preparation method for beta-amylase, which is less in resource consumption, high in yield (of about 75%), and high in activity (of 700000-900000 u / ml generally); and the obtained product is good in tolerance to temperature and pH value, and low in cost in case of being applied to maltose production.
Description
technical field [0001] The invention relates to a method for separating and purifying amylase in plants, in particular to a method for industrially producing high-activity beta-amylase. Background technique [0002] β-amylase (E.C3.2.1.2) is a kind of amylase, also known as 1,-3-maltosidase. β-amylase is widely found in barley, wheat, sweet potato, beans and other plants, and many microorganisms can produce β-amylase. β-amylase is used as a biocatalyst in the food industry and can be applied to the production and manufacture of maltose, beer, bread, etc. The use of β-amylase in food snacks can prevent starch aging, and the production of maltose for injection in the pharmaceutical industry can be used as a digestive agent together with α-amylase. [0003] In recent years, the use of β-amylase as a biocatalyst to produce maltose syrup has made up for the shortage of sugar in my country. With the advancement of science and technology, people are very active in research on hi...
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