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Aspergillus niger capable of highly yielding compound enzyme for feed and application thereof

A compound enzyme for feed, Aspergillus niger technology, applied in the direction of enzymes, lyases, hydrolytic enzymes, etc., can solve the problems of popularization and application obstacles, difficulties in growth and synthesis, and affecting the effect of use, so as to achieve good economic benefits and feed The effect of compound enzyme with less impurities and improved digestibility

Active Publication Date: 2014-04-23
HUNAN HONGYING BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantage is that the cost is high, and it is not easy to mix the enzyme preparation with high enzyme activity and low dosage with a large amount of feed when used, which seriously affects the effect of use, which increases the cost of use virtually, and makes many operators It is prohibitive and brings obstacles to promotion and application
2. Multi-strain mixed fermentation, natural complementary enzyme system method: use several strains for mixed fermentation, and the complementary enzyme production method to obtain a compound enzyme product that is closer to the compound method, and then only need a little adjustment to get the product, but multi-bacteria The technical difficulty of species fermentation is very large, because of the respective physiological characteristics of different strains, the mixed culture will inevitably affect each other, and it is difficult to make them grow on the same medium and synthesize different enzymes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Isolation, purification and identification of the Aspergillus niger DM-18 original strain of high-yield feed compound enzyme of embodiment 1

[0029](1) Strain isolation: Collect 10g of humus soil and rice straw mixed soil sample from Yangyou Township, Jin City, Hunan Province, add it to a small triangular flask containing 100mL of cooled sterile water with glass beads, shake at 200r / min, and shake Mix evenly for 30 minutes, then place in an 80°C water bath for 10 minutes, shake the Erlenmeyer flask constantly to mix the soil sample, let it stand for 5 minutes, draw 100 μL of the supernatant, and gradually dilute to 10 -1 ~10 -9 Concentration, select 10 -3 , 10 -4 , 10 -5 , 10 -6 Draw 0.2ml of the diluted solution and spread it on the PDA plate. After 4-5 days of constant temperature cultivation at 28°C, the colony will cover the entire plate.

[0030] (2) Strain purification: According to the morphological characteristics of the primary colony, a small amount of m...

Embodiment 2

[0037] The screening method of the bacterial strain of embodiment 2 high-yield compound enzymes for feed, comprises the following steps

[0038] 1) Slant culture: the isolated and purified Aspergillus niger original strain HYX0022 in Example 1 was streaked and inoculated on the slant medium, and cultured at 28-33° C. for 3-4 days until the mycelia matured and produced spores.

[0039] The composition of the slant medium is as follows: 200g of potatoes (boiled juice), 20g of glucose, 20g of agar, 1000mL of distilled water, pH value 5.8, sterilized at 121°C for 20min;

[0040] 2) Preparation of spore suspension: when Aspergillus niger produces spores, add 5 mL of sterile water to the inclined surface of the test tube, scrape off the spores, pour them into a sterilized triangular flask and add Tween-80 with 0.5-2% spore mass, Put the triangular flask in a shaker and vibrate for 1.5h to activate the spores; then centrifuge at 3500r / min for 15min, wash once with pH7.2 phosphate buf...

Embodiment 3

[0066] Example 3 The culture medium optimization of the starting strain for high-yield feed compound enzyme screening

[0067] (1) Strains

[0068] The starting strain is the DM-18 strain with the best screening results. After 96 hours of fermentation, the crude enzyme liquid of the fermentation broth contains a variety of enzymes, among which the protease activity is 6300U / ml, the mannanase activity is 1350U / ml, and the α -Galactase activity 1100U / ml, pectinase activity 900U / ml.

[0069] (2) Culture medium optimization

[0070] Optimizing the slant culture medium, plate culture medium, seed culture medium and fermentation medium in the process of strain screening, and fermenting for 96 hours under the same fermentation conditions before optimization. After testing, the crude enzyme liquid of the fermentation broth contains a variety of enzymes, Among them, protease activity 600U / ml, mannanase activity 1500U / ml, α-galactosidase activity 1200U / ml, pectinase activity

[0071]...

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Abstract

The invention discloses aspergillus niger capable of highly yielding a compound enzyme for a feed and an application thereof, and belongs to the technical field of microbial fermentation. The strain of the aspergillus niger specifically is aspergillus niger DM-18 which is collected in the China Center for Type Culture Collection with the collection number of CCTCC M2013541. A crude enzyme which is obtained through culture medium optimization and fermentation for 72-96 hours contains a plurality of kinds of enzymes, wherein the activity of protease is 6500U / ml, the activity of mannase is 1500U / ml, the activity of alpha-galactase is 1200U / ml and the activity of pectase is 1000U / ml. The optimum pH value for the activity of each enzyme in the compound enzyme ranges from 2 to 7, and is suitable for the digestion and absorption environment of the gastrointestinal tract of the animal; the compound enzyme for feed can be widely applied to the feeds for beasts and birds, is capable of reducing the antinutritional factors and increasing the digestibility, and has excellent economic benefits; simultaneously, the compound enzyme is also advantageous for protecting the economic environment and promoting healthy development of the animal husbandry.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, in particular to a high-yield feed compound enzyme Aspergillus niger and its application. Background technique [0002] A large number of studies at home and abroad have shown that adding enzyme preparations to feed can promote the digestion and absorption of nutrients by animals, improve feed utilization, and enhance animal disease resistance. Its main functions are as follows: (1) Supplement animal Insufficient endogenous enzymes improve feed digestibility and utilization. The enzyme preparation promotes the digestion and absorption of carbohydrates and other nutrients in the digestive tract, improves the conversion efficiency of feed, and is beneficial to the growth of animals. (2) Eliminate anti-nutritional factors in feed, reduce the viscosity of chyme in the digestive tract of animals, improve digestive function, and facilitate the absorption of nutrients. (3) Improve the d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12N9/40C12N9/18C12N9/26C12N9/88C12N9/62C12R1/685
Inventor 李洪兵张锦杰朱永明孙明芳陈香
Owner HUNAN HONGYING BIOTECHNOLOGY CO LTD
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