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Method for preparing beta-mannanase by using natural Bacillus subtilis

The technology of Bacillus subtilis and mannanase is applied in the field of preparation of β-mannanase, which can solve the problems of limitation, unstable performance of genetically engineered bacteria, ecological safety, etc. The effect of increasing the amount of enzymes produced by fermentation

Active Publication Date: 2011-08-17
DAQING HUALI ENERGY BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of improving enzyme production, it is mainly to carry out research around the coding gene of β-mannanase and construct enzyme-producing functional bacteria through genetic engineering. Although higher yields have been obtained, due to the unstable performance and Issues such as ecological security make the large-scale promotion of this technology subject to certain restrictions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] With culture medium: konjac fine powder 2.5~5.0% (wt%), peptone 0.1% (wt%), KH 2 PO 4 0.1% (wt%), MgSO 4 0.01% (wt%) was used as the culture medium, and the Bacillus subtilis HSO121 was cultivated at pH: 7-7.2, 37°C, 120r / min. After culturing for 32 hours, the crude enzyme solution was obtained after centrifugation to remove cells. The maximum enzyme activity was determined to be 13.16U / mL when it was reacted with the enzyme solution at 55°C for 10 minutes.

[0021] Konjac powder (wt%)

Embodiment 2

[0023] With medium: konjac fine powder 3.5% (wt%), KNO 3 80~140mmol / L, peptone 0.1% (wt%), KH 2 PO 4 0.1% (wt%), MgSO 4 0.01% (wt%) was used as the culture medium, and the Bacillus subtilis HSO121 was cultivated at pH: 7-7.2, 37°C, 120r / min. After culturing for 32 hours, the crude enzyme solution was obtained after centrifugation to remove cells. The maximum enzyme activity was determined to be 9.68U / mL when it was reacted with the enzyme solution at 55°C for 10 minutes.

[0024] KNO 3 mmol / L

Embodiment 3

[0026] With medium: konjac fine powder 3.5% (wt%), KNO 3 100mmol / L, peptone 0.1%~1.0% (wt%), KH 2 PO 4 0.1% (wt%), MgSO 4 0.01% (wt%) was used as the culture medium, and the Bacillus subtilis HSO121 was cultivated at pH: 7-7.2, 37°C, 120r / min. After culturing for 32 hours, the crude enzyme solution was obtained after centrifugation to remove cells. The maximum enzyme activity was determined to be 14.31U / mL when it was reacted with the enzyme solution at 55°C for 10 minutes.

[0027] Peptone (wt%)

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Abstract

The invention provides a method for preparing beta-mannanase by using natural Bacillus subtilis, which comprises: culturing Bacillus subtilis HSO121 by using culture medium containing refined rivier giantarum rhizome powder and a composite nitrogen source, centrifuging the obtained culture liquid to remove thallus cells and thus obtaining coarse beta-mannanase liquid, wherein the mass concentration of refined rivier giantarum rhizome powder in the culture medium is 2.5 to 5.0 percent; and the composite nitrogen source comprises KNO3 at a concentration of 80 to 140mmol / L and peptone at a concentration of 0.1 to 1 weight percent. In the invention, the beta-mannanase is prepared by optimizing the formula of the culture medium and by using the natural Bacillus subtilis HSO121, so the beta-mannanase yield of the fermentation of the Bacillus subtilis HSO121 is improved and a good technical effect is produced.

Description

technical field [0001] The invention relates to the technology of producing enzymes by microorganisms, in particular to a preparation method of β-mannanase. Background technique [0002] Mannan is rich in sources, and different types of mannan exist in konjac flour, guar gum, locust bean gum, etc. Backbone mannan degrading enzymes include β-mannanase, β-glucosidase, β-mannosidase. Among them, the most important is β-mannanase. β-mannanase is a kind of endohydrolase that can hydrolyze mannan oligosaccharides and mannan polysaccharides containing β-1,4-D-mannosidic bonds. It belongs to hemicellulase and is widely found in animals and plants. and microorganisms. Among them, microorganisms are the main source of β-mannanase, which has the advantages of high activity and stable source. [0003] With the development of natural hemicellulose resources in recent years, the research and development of β-mannanase has entered a new climax, and has been widely used in food, medicin...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12R1/125
Inventor 牟伯中杨世忠刘金峰陈晓青
Owner DAQING HUALI ENERGY BIOLOGICAL TECH
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