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Method for fermentation production of beta-mannase

A technology of mannanase and solid fermentation, which is applied in the field of microbial fermentation, can solve problems such as high cost pressure, achieve the effects of improved production performance, good temperature resistance, and reduced production costs

Active Publication Date: 2013-04-17
GUANGDONG VTR BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as the price of production raw materials (soybean meal, etc.) continues to rise, the cost pressure of the enzyme preparation industry is increasing. Therefore, a solid fermentation medium that can reduce production costs and increase enzyme activity can enable enterprises to compete in the enzyme preparation industry. invincible

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  • Method for fermentation production of beta-mannase

Examples

Experimental program
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Effect test

preparation example Construction

[0058] 2.2 Preparation of enzyme solution to be tested

[0059] Accurately weigh about 1.000g of a certain amount of sample into a 50mL colorimetric tube, add 19mL of distilled water and shake in a vortex mixer until it is dissolved, soak at room temperature for 45min, and shake once every 15min during the extraction process. After leaching, take the supernatant as the test enzyme solution, which is diluted to an appropriate multiple for use.

[0060] 2.3 β-Mannanase activity determination

[0061] Add 2 mL of the substrate solution to 25 mL colorimetric tubes A and B. After preheating at 55°C for 5 minutes, add 0.5 mL of appropriately diluted enzyme solution to tube A, react accurately in a water bath at 55°C for 10 minutes, and add to tubes A and B. 2.5mL DNS reagent, add 0.5mL enzyme solution to tube B, immediately boil for 5min, quickly cool with ice water, add 5mL distilled water, shake well, measure the optical density at 520nm (tube B is blank control), Find the correspondin...

Embodiment 1 3

[0064] Example 1 Triangular flask solid fermentation β-mannanase 1

[0065] Base material (80g bran, 7g soybean meal, 13g palm meal), 0.7g konjac flour, (NH 4 ) 2 SO 4 2g, KH 2 PO 4 1g, water 153mL.

[0066] Triangle flask solid fermentation medium: 300mL flask 15g base material (including: bran 12g, soybean meal 1g, palm meal 2g), konjac flour 0.1g, (NH 4 ) 2 SO 4 0.3g, KH 2 PO 4 0.15g, 23mL tap water, pH 5.0.

[0067] Fermentation conditions: Sterilize at 121°C for 30min, inoculate 2mL spore suspension after cooling, mix well, culture at 32°C for 96h, and buckle the bottle once at 22h and 40h after inoculation.

[0068] The results of repeated 5 batches of Erlenmeyer flask fermentation test showed that the mature fermented koji material was dried and crushed in a blast oven at 45~50℃, and the average β-mannanase activity reached 26072U / g dry koji.

Embodiment 2 3

[0069] Example 2 Triangular flask solid fermentation β-mannanase 2

[0070] Triangle flask solid fermentation medium: 300mL flask 15g base material (including: bran 11g, soybean meal 1.5g, palm meal 2.5g), konjac flour 0.3g, (NH 4 ) 2 SO 4 0.45g, KH 2 PO 4 0.3g, 26mL tap water, pH7.0.

[0071] Fermentation conditions: Sterilize at 121°C for 30 minutes, inoculate 4 mL of spore suspension after cooling, mix well, cultivate at 33°C for 84 hours, and buckle the bottle once each 20h and 40h after inoculation.

[0072] Repeated 5 batches of Erlenmeyer flask fermentation test results showed that the mature fermented koji material was dried and crushed in a blast oven at 45~50℃, and the average β-mannanase activity reached 24867U / g dry koji.

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Abstract

The invention relates to the field of microbial fermentation, and in particular to a method for fermentation production of beta-mannase. The method comprises the step of expressing beta-mannase by aspergillus usamii By247 through fermentation. According to the mannose produced by the invention, palm meal is used as one of the fermenting materials, and animal experimental results show that the mannase can decompose antinutritional factors in the palm meal in unconventional feed well and remarkably improve the production performance of animals. The temperature resistance, the feed and material decomposing capacity of the mannase and the result of animal experiments are better than those of other merchant beta-mannase.

Description

Technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for fermentative production of β-mannanase. Background technique [0002] β-mannanase is a type of endonuclease that can hydrolyze β-1,4-D-mannosidic bonds, and belongs to the class of hemicellulase. It has the effect of general non-starch polysaccharidase-eliminates the anti-nutritional effect of β-mannan, promotes the digestion and absorption of nutrients by animals, and improves feed conversion and energy utilization; recent studies have found that β-mannan Enzyme is also a multifunctional growth-promoting agent. It can promote the secretion of insulin-like growth factor IGF-I, promote protein synthesis, and increase lean meat rate; its product, mannose oligosaccharides, can improve the intestinal microbial environment and improve animal immune function . [0003] β-mannanase is widely present in nature, and β-mannanase activity is found in some lower animals, plants ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12R1/66
Inventor 谢志恒王敏崔细鹏周平发翁晓辉史宝军
Owner GUANGDONG VTR BIO TECH
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