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Preparation method of high-activity composite enzyme for feed

A feed compound enzyme and high-activity technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve problems such as obstacles to popularization and application, difficulties in growth and synthesis, and impact on the effect of use, and achieve enhanced The body is functional, suitable for industrial production, and has high vitality

Active Publication Date: 2015-02-11
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

[0054] Example 1 Feed Compound Enzyme Enzyme Activity Determination

[0055] 1) Definition of enzyme activity unit:

[0056] Protease enzyme activity unit definition: under the conditions of 40°C and pH 5.5, the amount of enzyme that hydrolyzes casein for 1 minute to produce 1 μmol of tyrosine is 1 enzyme activity unit, expressed in U / ml.

[0057] Definition of mannanase activity unit: at 40°C and pH 5.5, the amount of enzyme required to hydrolyze and release reducing sugar equivalent to 1 μmol mannose from a locust bean gum solution with a concentration of 5 mg / ml in 1 min is 1 enzyme Vitality unit, expressed in U / ml.

[0058] Definition of α-galactosidase activity unit: at 40°C and pH 5.5, the amount of enzyme required to decompose p-nitrophenol-α-D-galactopyranose to generate 1 μmol p-nitrophenol in 1 minute is 1 α-galactose The enzyme activity unit of lactosylase is expressed in U / ml.

[0059] Definition of pectinase activity unit: at 40°C and pH 5.5, the amount of enzy...

Embodiment 2

[0063] A method for preparing a compound enzyme for high-activity feed comprises the following steps:

[0064] (1) Activation of strains

[0065] Inoculate the well-preserved slant strain of Aspergillus niger DM-18 on the slant medium, culture at 28°C for 36 hours to activate the strain, and activate twice in this way;

[0066] The slant medium consists of: casein 4g, dipotassium hydrogen phosphate 2g, magnesium chloride 0.6g, potassium chloride 0.8g, ferrous sulfate 0.02g, glucose 20g, agar 20g, Chinese herbal medicine powder 5g, distilled water 1000mL, pH value 5.8 , sterilized at 121°C for 20 minutes;

[0067] The preparation method of described Chinese herbal medicine powder is as follows:

[0068] Weigh 20 parts of Radix Astragali; 10 parts of Codonopsis; 10 parts of Bupleurum; Keep it for 2 hours, add a mixture of ethanol and propanol twice the weight of the mixed material, control the temperature to 60° C., keep it for 3 hours, and filter; the filtrate is concentrate...

Embodiment 3

[0090] A method for preparing a compound enzyme for high-activity feed comprises the following steps:

[0091] (1) Activation of strains

[0092] Inoculate the well-preserved slant strain of Aspergillus niger DM-18 on the slant medium, culture at 34°C for 48 hours to activate the strain, and activate it 4 times;

[0093] The slant medium consists of: casein 4g, dipotassium hydrogen phosphate 2g, magnesium chloride 0.6g, potassium chloride 0.8g, ferrous sulfate 0.02g, glucose 20g, agar 20g, Chinese herbal medicine powder 20g, distilled water 1000mL, pH value 5.8 , sterilized at 121°C for 20 minutes.

[0094] The preparation method of described Chinese herbal medicine powder is as follows:

[0095] Weigh 30 parts of Radix Astragali; 18 parts of Codonopsis; 15 parts of Bupleurum; Keep it for 4 hours, add a mixture of ethanol and propanol 3 times the weight of the mixed material, control the temperature to 78° C., keep it for 4 hours, filter; the filtrate is concentrated in vac...

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Abstract

The invention discloses a preparation method of a high-activity composite enzyme for feed, belonging to the technical field of preparation of an enzyme preparation for feed. An Aspergillus niger DM-18 strain for a high-yield composite enzyme for feed, which is used as an initial strain, is subjected to liquid submerged mixed fermentation in an optimized culture medium under specific fermentation conditions by an improve fermentation technique to obtain the composite enzyme for feed, which has the advantages of complete enzymatic system, high enzyme activity, high temperature resistance and wide pH value resistance range. The prepared crude enzyme fermentation liquid contains multiple enzyme activities, wherein the proteinase activity is 6500-6700 U / ml, the mannase activity is 1500-1700 U / ml, the alpha-galactase activity is 1200-1300 U / ml, and the pectinase activity is 1000-1200 U / ml. The enzymes still have higher enzyme activity (up to 80% or above) at 30-75 DEG C under the pH value of 2-7; and thus, the composite enzyme is more suitable for the demands of a processing technique in feed industry, and the processed feed has low loss of enzyme activity.

Description

technical field [0001] The invention belongs to the technical field of enzyme preparation, in particular to a preparation method of a high-activity compound enzyme for feed. 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 distribution...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00A23K1/165C12N9/88C12N9/62C12N9/42C12N9/40C12N9/26C12N9/18C12R1/685
Inventor 李洪兵张锦杰李贵骏易继云刘文明
Owner HUNAN HONGYING BIOTECHNOLOGY CO LTD
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