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Preparation method of amylase

A technology of amylase and starch, applied in the direction of biochemical equipment and methods, enzymes, hydrolytic enzymes, etc., to achieve high yield and improve the effect of purity

Active Publication Date: 2014-05-14
DUODUO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, many studies and patents have reported the extraction of α-amylase from microorganisms [Chen Bo, Li Dali, Yang Shulin. Screening of acid α-amylase production strains, enzyme purification and enzyme properties research [J]. Food Science , 2005, 26(5): 119-122. Wang Hongmei. Study on solid-state fermentation conditions and enzyme separation and purification of acid-resistant α-amylase produced by Aspergillus niger PZ301 [D]. Anhui, Anhui Agricultural University, 2011. Zeng Lijuan. Separation and purification of amylase produced by Bacillus strains and preliminary research on the starch binding domain CBM25 [D]. Guangxi, Guangxi University, 2010. A strain of A-amylase inhibitor production bacteria and the preparation method and method of A-amylase inhibitor Should. CN 101139569. ⑤ The method for the separation and purification of α-amylase by two-phase extraction. CN 102220300B.], there are also many patents disclosing the extraction of β-amylase [soybean β-amylase] from plants such as bran, soybean and potato. Amylase preparation process. CN 1088754C. Preparation method of liquid β-amylase preparation. CN 101323849 B. Method for extracting β-amylase from potato cell tissue. CN 101451127 B. Extracting liquid β-amylase from bran method.CN101979529 B.], but extracting amylase from malt is also rarely reported

Method used

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  • Preparation method of amylase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 A method for extracting amylase from barley malt

[0048] The method for extracting amylase from barley malt comprises the following steps:

[0049] (1) Crushed barley malt

[0050] The barley malt is placed in a pulverizer to pulverize, and the pulverized product is passed through an 80-mesh sieve, and the undersize is collected. Weighing, ration packaging, and marking the name, weight, batch number and other content for future use.

[0051] (2) Leaching

[0052] (1) Extraction preparation: Add 0.25kg of sodium metabisulfite, 0.25kg of sodium pyrosulfite, 0.25kg of sodium sulfite, 0.25kg of sodium thiosulfate and 0.05kg of potassium sorbate to 100L of purified water, mix well and set aside.

[0053] (2) Amylase extraction: Weigh 10kg of barley malt crushed material into the extraction solution prepared in step (1), add 0-5g of cellulose compound enzyme, mix well, heat and control the temperature under normal pressure at 20-45°C, adjust the pH value to 5, ...

Embodiment 2

[0071] Example 2 Effect of the amount of cellulose compound enzyme on the activity of amylase in the concentrated enzyme solution

[0072] Steps:

[0073] (1) The crushing of barley malt is the same as in Example 1.

[0074] (2) Leaching

[0075] (1) Extraction preparation: Add 0.25kg of sodium metabisulfite, 0.25kg of sodium metabisulfite, 0.25kg of sodium bisulfite, 0.25kg of sodium thiosulfate and 0.05kg of potassium sorbate to 100L of purified water, mix well and set aside.

[0076] (2) Amylase extraction: Weigh 10 kg of barley malt pulverization and add it to the extract prepared in step (1), add 0, 1, 2, 3, 4, 5 g of cellulose compound enzyme (β-glucan Enzyme activity not less than 8000U / g, cellulase activity not less than 200U / g, neutral protease activity not less than 500U / g, xylanase activity not less than 3000U / g), divided into six groups for testing. After fully mixing, under normal pressure, heat and control the temperature to 30°C, adjust the pH value to 5, and...

Embodiment 3

[0121] Example 3 Effect of Extraction Temperature on Amylase Activity in Concentrated Enzyme Liquid

[0122] Steps:

[0123] (1) The crushing of barley malt is the same as in Example 1.

[0124] (2) Leaching

[0125] (1) Extraction preparation: Add 0.25kg of sodium metabisulfite, 0.25kg of sodium metabisulfite, 0.25kg of sodium bisulfite, 0.25kg of sodium thiosulfate and 0.05kg of potassium sorbate to 100L of purified water, mix well and set aside.

[0126] (2) Amylase extraction: Weigh 10kg of barley malt ground into the extract prepared in step (1), add 3g of cellulose complex enzyme (β-glucanase activity is not less than 8000U / g, cellulase Activity of not less than 200U / g, activity of neutral protease not less than 500U / g, activity of xylanase not less than 3000U / g), after mixing thoroughly, heat and control the temperature at 20°C under normal pressure , 25°C, 30°C, 35°C, 40°C, 45°C, divided into six groups for testing. Adjust the pH value to 5, and extract with stirri...

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Abstract

The invention discloses a preparation method of amylase. With barley malt as a raw material, the method comprises the following steps: grinding of barley malt, extraction of amylase, preliminary filtration of the extract liquid, enrichment of the amylase, precipitation of the extract liquid, fine filtration of the extract liquid, ultrafiltration concentration, blending, secondary fine filtration and aseptic packaging. By adopting the method disclosed by the invention, the enzyme activity of the prepared amylase is not lower than 700,000U / ml, the purity is high, the stability is good, and the application prospect is broad.

Description

technical field [0001] The invention relates to a method for preparing amylase, in particular to a method for extracting amylase from barley malt. Background technique [0002] The main storage substance in malt is starch, including 20% ​​to 40% amylose and 60% to 80% amylopectin. At the same time, amylases closely related to starch degradation in malt include α-amylase, β-amylase and limit dextrinase [Wolfgang K. Practical Technology of Beer Technology [M]. Beijing: China Light Industry Press, 1998; Macgergor A.W. , Bazin S.L., Macri L.J., et al.. Modeling the contribution of alpha-amylase, beta-amylase and limit dextrinase to starch degradation during mashing [J]. Journal of Cereal Science, 1999, 29: 161-169]. [0003] α-Amylase is an endo-amylase. It can hydrolyze the α-1,4 glucoside bond in the starch molecular chain at will, but not the α-1,6 glucoside bond. When α-amylase acts on amylose, it generates maltose, glucose and small molecule dextrin; when it acts on amyl...

Claims

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Application Information

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IPC IPC(8): C12N9/32C12N9/24
CPCC12N9/2422C12N9/2425C12Y302/01001C12Y302/01002
Inventor 王发善韩振远褚弘斌陈玉忠
Owner DUODUO PHARMA
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