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Preparation of beta-mannase enriched compound enzyme as well as strain and application thereof

A technology of mannanase and compound enzymes, which is applied in the field of compound enzymes, can solve problems such as poor compound enzyme systems, and achieve the effect of complete enzyme components, rich enzyme systems, and inhibited growth

Active Publication Date: 2020-09-01
HANGZHOU BIOCOM BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention proposes the preparation of a compound enzyme rich in β-mannanase and its preparation and application, which solves the problem of the poor compound enzyme system caused by single preparation of the enzyme preparation in the prior art

Method used

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  • Preparation of beta-mannase enriched compound enzyme as well as strain and application thereof
  • Preparation of beta-mannase enriched compound enzyme as well as strain and application thereof
  • Preparation of beta-mannase enriched compound enzyme as well as strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A kind of preparation that is rich in the complex enzyme of β-mannanase, comprises the steps:

[0040] 1 strain activation

[0041] The Aspergillus niger BAK200310 strain preserved by the company was inoculated on the slant of the sterilized first-level test tube for culture, and cultured at 28-32°C for 6-10 days. The medium formula was: glucose 1-2%, potato juice 20-30%, Agar powder 1.2-1.8%.

[0042] 2 strain expansion

[0043] Inoculate the slant seeds of the primary test tube obtained in step 1 into the solid medium of the secondary Erlenmeyer flask for cultivation. Transfer the slant spores to 150-200mL sterile water in an ultra-clean workbench, shake and mix well, so that the spores are fully suspended in the sterile water. Then inoculate on the solid medium of the secondary Erlenmeyer flask by 2-3mL / bottle solid seed amount to ferment. Cultivate at 28-32°C for 4-6 days. The medium formula is: 100% bran, 5%-10% ammonium sulfate, 45%-55% initial moisture, and ...

Embodiment 2

[0048] The solid medium in embodiment 1 can be selected for use in the following ratio preparation:

[0049] Bran 70-80%, corn cob flour 20%-30%, ammonium sulfate 1-2%, magnesium sulfate 0.05%-0.1%, calcium carbonate 0.5%-1%, dipotassium hydrogen phosphate 0.2%-0.3%, initial Moisture 60%-65%;

[0050] or,

[0051] Bran 60-70%, soybean meal 10%-20%, corncob powder 5%-10%, calcium carbonate 1%-2%, ammonium chloride 1-2%, magnesium sulfate 0.05%-0.1%, manganese sulfate 0.02%-0.03%, dipotassium hydrogen phosphate 0.2%-0.3%, initial moisture 60%-65%.

[0052] Strain activation can also be replaced by the following medium:

[0053] Animal tissue pepsin hydrolyzate and tryptone equal mixture 1%, glucose 4%, agar powder 1.2-1.8%, pH 5.4-5.8.

[0054] All the other conditions are constant, all can realize the object of the present invention.

Embodiment 3

[0056] Enzyme protein detection analysis result (for embodiment 1 fermentation product) in compound enzyme

[0057] Dry and pulverize the fermented product after solid fermentation to make a compound enzyme sample, add 8M urea solution to extract the protein components in the compound enzyme sample, use LC-MS to identify the protein components, first quantify the protein, and then reduce the alkane The three-dimensional structure of the protein is opened by hydrolysis, the peptides are extracted after enzymatic hydrolysis, and the mass spectra of these peptides are obtained by using mass spectrometry technology. Finally, the protein in the sample is identified by using protein identification software. The obtained results were searched and compared in the protein library (Aspergillus niger library), and the comparison results were further analyzed.

[0058] According to the test and analysis results, the compound enzyme sample provided by our company has many kinds of enzyme p...

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Abstract

The invention provides a compound enzyme rich in beta-mannase as well as preparation and application thereof. The cultured fermentation product contains beta-mannase, beta-glucanase, acidic protease,xylanase, pectinase and alpha-galactosidase. Aspergillus niger is named as Aspergillus niger BAK200310, and the preservation number is CGMCC No. 19616. The fermented compound enzyme is extremely richin zymoprotein types, contains various zymoprotein components, and almost covers all components in conventional enzymes; different enzymes perform respective functions, and different components of onesame enzyme cooperate with each other to jointly exert a greater enzymatic hydrolysis effect. The compound enzyme is rich beta-mannase and complete in enzyme components, and the enzyme activity of the compound enzyme can reach 12170U / g according to national standard detection.

Description

technical field [0001] The invention relates to the field of compound enzymes, in particular to the preparation of a compound enzyme rich in β-mannanase and its preparation and application. Background technique [0002] With the improvement of people's living standards, people's demand for livestock and poultry products has increased year by year, which has driven the rapid development of aquaculture and feed industries. The acceleration of urbanization and large-scale breeding has led to increasingly prominent problems of feed resources and environmental protection in animal husbandry. To solve such problems, it is necessary to develop new feed resources and make full use of existing resources, but also to seek ways to improve nutrient utilization efficiency and reduce environmental pollution. Enzyme is a kind of protein with biocatalytic activity. The feed for livestock and poultry is mainly plant-based raw materials, including corn, wheat, soybean meal, cotton meal, etc....

Claims

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Application Information

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IPC IPC(8): C12N9/24C12N9/42C12N9/50C12N9/18C12N9/40C12N9/88C12N1/14A23K20/189A23K50/75C12R1/685
CPCC12N9/2494C12N9/2437C12N9/50C12N9/248C12N9/2465C12N9/18C12N9/2402C12N9/88C12N1/14A23K20/189A23K50/75C12Y302/01078C12Y302/01022C12Y301/01011C12Y302/01015C12Y402/02002C12R2001/685C12N1/145Y02P60/87
Inventor 王云龙吴勃徐永雷王天珍刘广晓
Owner HANGZHOU BIOCOM BIOLOGICAL TECH
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