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Beta-glucosidase preparing method

A glucosidase and temperature control technology, applied in the field of preparation of β-glucosidase, can solve the problems of high cost and difficulty in large-scale production of non-food energy projects, achieve better enzymatic hydrolysis effect, improve mixing uniformity, reduce The effect of small losses

Active Publication Date: 2013-04-17
安徽丰原集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problem that current non-grain energy projects are difficult to put into large-scale production due to high cost, and to provide a method to produce β-glucosidase by using microorganisms

Method used

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  • Beta-glucosidase preparing method

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Preparation of PDA slant medium: 200g peeled potatoes, chopped, add 1000ml water to boil for 30min, filter out the potato residue, add 20g sucrose to the filtrate, dissolve and set the volume to 1000ml, add 20g agar, boil to dissolve, then pack into test tubes, Cotton plugs, 7 in a group, wrapped with kraft paper, sterilized at 121°C for 20 minutes, waited for about 60-70°C, placed on a slope to cool and solidify, then put it in a 30°C incubator for 24 hours, check that there is no contamination use.

[0021] Take a ring from the refrigerated Aspergillus niger test tube, apply it to the inclined surface of the PDA test tube, place it in a constant temperature incubator at 30°C, cultivate it for 7-8 days, and put it in a 4°C refrigerator to refrigerate, then it can be used as the inclined plane seed.

Embodiment 2

[0023] The solid fermentation medium used in the method of the present invention contains per kilogram: corn cob 180g, bran 120g, ammonium sulfate 50g, urea 20g, magnesium sulfate 0.5g, potassium dihydrogen phosphate 0.1g, water 630g, and mix uniformly.

[0024] Put 120 g of the culture medium obtained above into a 1000 ml Erlenmeyer flask, seal it with a cotton plug, tie the mouth with kraft paper, and sterilize at 121° C. for 60 min. Take a loop of spores on the slant of the PDA medium test tube to inoculate, and culture statically at 30°C for 4-6 days. After the bran koji is covered with dark brown mold spores and mature, it is ready for use.

Embodiment 3

[0026] The solid fermentation medium contains per kilogram: 180g corn cob, 120g bran, 50g ammonium sulfate, 20g urea, 0.5g magnesium sulfate, 0.1g potassium dihydrogen phosphate, 630g water, and mix well.

[0027] Fill the solid fermentation medium with 10kg in a 1m×1m×0.1m square pan, cover the lid, sterilize at 121°C for 60min, cool it and inoculate it with black spores of the mold in the triangular flask, the inoculation amount is 2% of the weight of the solid medium Mix and inoculate with a mixer, then distribute 6kg per plate, and culture at 30°C. Water and moisturize the culture medium on the first day, and then continue to stand still for 3-4 days, dry and pulverize the cultured enzyme at 50°C and pass through a 200-mesh sieve to make enzyme powder.

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Abstract

The invention relates to a beta-glucosidase preparing method. A native cellulose matter is used as a carbon source to manufacture a solid fermentation culture medium, and beta-glucosidase is prepared by mechanical stirring inoculation and solid disc type fermentation. The solid fermentation culture medium comprises the following components in portion by weight: 160 portions of native cellulose matter, 140 portions of bran, 30-50 portions of ammonium sulphate, 10-30 portions of urea, 0.2-0.5 portion of magnesium sulfate, 0.05-0.1 portion of monopotassium phosphate, and 550-650 portions of water. The fermentation by using a solid fermentation tank is replaced with the solid disc type fermentation, therefore, the invention has thorough disinfection and sterilization, is easy to treat local contamination, reduces the loss caused by contamination and is beneficial to scale production; during the fermentation period, the heat in previous fermentation period is fast to generate, therefore, the heat can be controlled to generate to be used as the heat required by the growth, and the electricity consumption is saved. The beta-glucosidase has 1200 IU / g of enzyme activity and can be producedin a large-scale industrial way through cost accounting.

Description

technical field [0001] The invention relates to the field of microorganism production and manufacturing, in particular, to a preparation method of β-glucosidase. Background technique [0002] β-glucosidase (β-glucosidase, EC3.2.1.21), belongs to hydrolase, also known as β-D-glucoside hydrolase, alias gentiobiase, cellobiase and amygdalinase, It is a kind of cellulose complex enzyme system. It belongs to cellulase, which can catalyze the hydrolysis of glycosidic bonds between aryl or hydrocarbon groups and sugar groups to generate glucose. β-glucosidase is widely used in straw alcohol and other industries. Its function is to degrade cellobiose produced by straw under the action of cellulase. Cellobiose is an inhibitor of cellulase activity. With the increase of cellobiose reduced, the efficiency of cellulase hydrolysis was improved. There are many substrates that can be hydrolyzed by β-glucosidase, such as daidzin and amygdalin, which can be widely used in the food and dru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12R1/685
Inventor 李荣杰薛培俭穆晓玲李维理
Owner 安徽丰原集团有限公司
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