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,

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

Example Embodiment

[0038] Example 1

[0039] The preparation of a composite enzyme rich in β-mannanase includes the following steps:

[0040] 1 Strain activation

[0041] The Aspergillus niger BAK200310 strain kept by the company was inoculated into the sterilized primary test tube slant and cultivated 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] The slant seeds of the primary test tube obtained in step 1 were inoculated into the secondary Erlenmeyer flask solid medium 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 sterile water. Then 2-3 mL / bottle of solid seeds were inoculated onto the secondary Erlenmeyer flask solid medium for fermentation. Incubate at 28-32°C for 4-6 days. The medium formula is: bran 100%, ammonium sulfate 5%-10%, initial water content 45%-55%, and the amount of medium ...

Example Embodiment

[0047] Example 2

[0048] The solid medium in Example 1 can be prepared in the following proportions:

[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 powder 10%-20%, corn cob 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 content 60%-65%.

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

[0053] An equal mixture of animal tissue pepsin hydrolysate and tryptone 1%, glucose 4%, agar powder 1.2-1.8%, pH 5.4-5.8.

[0054] If the other conditions remain unchanged, the objective of the present invention can be achieved.

Example Embodiment

[0055] Example 3

[0056] Results of detection and analysis of enzyme protein in complex enzyme (for the fermentation product of Example 1)

[0057] Dry and pulverize the fermentation product after solid fermentation to make a composite enzyme sample, add 8M urea solution to extract the protein components in the composite enzyme sample, use LC-MS to identify the components of the protein, first quantify the protein, and reduce the alkane Grouping opens the three-dimensional structure of the protein, extracts the peptides after enzymolysis, uses mass spectrometry to obtain the mass spectra of these peptides, and finally uses protein identification software to identify the proteins in the sample. The results are searched and compared in the protein library (Aspergillus niger library), and the comparison results are further analyzed.

[0058] The test and analysis results show that there are many types of enzyme proteins (nearly 50) in the composite enzyme samples provided by the compa...

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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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