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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, and achieve the effects of improved production performance, improved enzyme activity, and good temperature resistance

Active Publication Date: 2015-01-07
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
  • Method for fermentation production of beta-mannase
  • Method for fermentation production of beta-mannase

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Experimental program
Comparison scheme
Effect test

preparation example Construction

[0058] 2.2 Preparation of the enzyme solution to be tested

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

[0060] 2.3 β-mannanase activity assay

[0061] Add 2mL of substrate solution to 25mL colorimetric tubes A and B, preheat at 55°C for 5min, then add 0.5mL of appropriately diluted enzyme solution to tube A, react in a water bath at 55°C for 10min, add Add 0.5mL of enzyme solution to 2.5mL of DNS reagent in tube B, immediately boil for 5min, add 5mL of distilled water after rapid cooling with ice water, oscillate and shake well, measure the optical density at 520nm (tube B is the blank control), and extra...

Embodiment 1 3

[0064] Embodiment 1 Erlenmeyer flask solid fermentation β-mannanase 1

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

[0066] Triangular flask solid fermentation medium: 300mL triangular 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, tap water 23mL, pH 5.0.

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

[0068] The results of repeated 5 batches of triangular flask fermentation tests showed that the average enzyme activity of β-mannanase reached 26072U / g dry koji after the mature fermented koji material was dried and pulverized in a blast oven at 45-50°C.

Embodiment 2 3

[0069] Embodiment 2 Erlenmeyer flask solid fermentation β-mannanase 2

[0070] Erlenmeyer flask solid fermentation medium: 300mL Erlenmeyer 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, tap water 26mL, pH7.0.

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

[0072] The results of repeated 5 batches of triangular flask fermentation tests showed that the average enzyme activity of β-mannanase reached 24867U / g dry koji after the mature fermented koji material was dried and pulverized in a blast oven at 45-50°C.

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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 fermenting and producing β-mannanase. Background technique [0002] β-mannanase is a class of endonucleases capable of hydrolyzing β-1,4-D-mannosidic bonds, belonging to the hemicellulase class. It has the function of general non-starch polysaccharidase - eliminate the anti-nutritional effect of β-mannan, promote the digestion and absorption of nutrients by animals, improve feed conversion rate and energy utilization rate; many recent studies have found that β-mannan The enzyme is also a multifunctional growth-promoting agent, which can promote the secretion of insulin-like growth factor IGF-I, promote protein synthesis, and increase lean meat percentage; its product mannan oligosaccharide can improve the intestinal microbial environment and improve animal immune function . [0003] β-mannanase exists widely in nature, and β-mannanase activity is found in some...

Claims

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Application Information

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Patent Type & Authority Patents(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