Preparation of enzyme preparation with acidic proteinase as main component as well as strain and application of enzyme preparation

A technology of acid protease and enzyme preparation, which is applied in the field of preparation of enzyme preparations, can solve the problems of many protein components, inappropriateness, and high energy consumption, and achieve the effects of many types of enzymes, considerable enzyme activity, and stable growth

Active Publication Date: 2020-08-28
HANGZHOU BIOCOM BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011]2. The enzyme protein detection method of enzyme preparation is mature, but the application is relatively single, usually for the analysis of a certain protein
This method is not suitable for solid fermentation products with many protein components and difficult separation and purification, and a more suitable method needs to be found
[0012]3. At present, there is no report on the sys

Method used

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  • Preparation of enzyme preparation with acidic proteinase as main component as well as strain and application of enzyme preparation
  • Preparation of enzyme preparation with acidic proteinase as main component as well as strain and application of enzyme preparation
  • Preparation of enzyme preparation with acidic proteinase as main component as well as strain and application of enzyme preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] A preparation of an enzyme preparation based on acid protease, comprising the following steps:

[0058] 1. Primary slant culture

[0059] The strain Asperijilus niger (Asperijilus niger) BAK200389 was inoculated on the potato juice glucose slant medium, and cultured at 28° C. to 31° C. for 5 to 7 days to obtain the slant that germinated spores.

[0060] 2. Secondary Liquid Seed Preparation

[0061] In the ultra-clean workbench, scrape the spore mycelium mixture on the slope in step 1, inoculate it into the liquid medium in the shaker flask, and culture it with shaking on a shaker. When the seed bacteria liquid in the bottle becomes viscous, the culture is terminated and stored in the refrigerator.

[0062] The shaker culture conditions are as follows: 28°C-30°C, rotation speed 180-220r, culture period 1-3d.

[0063] The shake flask volume of described bran culture medium is 250 / 500mL.

[0064] The liquid medium is formulated according to the following weight ratios ...

Embodiment 2

[0092] A preparation of an enzyme preparation based on acid protease, comprising the following steps:

[0093] 1. Primary slant culture

[0094]The fungus Asperijilus niger (Asperijilus niger) CGMCC No.19615 was inoculated on the modified Martin slant medium, and cultured at 28° C. for 5-7 days to obtain a spore-forming slant.

[0095] The medium components: 5.0 g of peptone, 1.0 g of dipotassium hydrogen phosphate, 2.0 g of yeast extract powder, 0.5 g of magnesium sulfate, 20.0 g of glucose, 1000 ml of water, and the pH is adjusted to 6.8.

[0096] 2. Secondary Liquid Seed Preparation

[0097] In the ultra-clean workbench, scrape the spore mycelium mixture on the slope in step 1, inoculate it into the liquid medium in the shaker flask, and culture it with shaking on a shaker. When the seed bacteria liquid in the bottle becomes viscous, the culture is terminated and stored in the refrigerator.

[0098] The shaker culture conditions are as follows: 28°C-30°C, rotation speed ...

Embodiment 3

[0127] Protein detection methods and assay types:

[0128] Using proteomics analysis method, using LC-MS for protein identification, after software analysis, identified 38 enzyme proteins, a total of 7 categories, mainly acid protease, pectinase, xylanase, amylase, Cellulase, mannanase, glucosidase, glucosidase, galactosidase, ferulic esterase, carboxypeptidase, phosphatase and other enzymes are supplemented. Partzyme proteins are listed below:

[0129] Aspergillus 1 (aspartic protease pepA), aspergillus 2 (acid protease A), carboxypeptidase, serine carboxypeptidase F, pectin esterase A, endogalacturonase A (fruit Gelase A), α-L-arabinofuranosidase axhA, α-L-arabinofuranosidase A, α-xylosidase A, β-xylosidase A, feruloesterase B, esterase A , 1,4-β-endoxylanase C (xylanase C), α-L-arabinofuranosidase B, 1,4-β-endoxylanase A (xylanase A), 1,4-β-endoxylanase B (xylanase B), α-amylase type A 1 / 2, glucoamylase, 1,4-β-D-glucan Cellobiohydrolase A, 1,4-β-D-glucan Cellobiohydrola...

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Abstract

The invention provides preparation of an enzyme preparation with acidic proteinase as a main component as well as a strain and application of the enzyme preparation. The multi-enzyme series acidic proteinase preparation which utilizes the acidic proteinase as the main component, utilizes multiple associated enzymes including pectinase, xylanase, amylase, cellulase, mannase, glucanase, glucosidase,galactosidase, feruloyl esterase, carboxypeptidase and phosphatase and is rich in natural complex enzymes is obtained by culture. Aspergillus niger is named Aspergillus niger BAK200389, and the preservation number of the Aspergillus niger is CGMCC No.19613. According to the used strain, the strain can produce multiple enzymes, and has the characteristics of stable growth, more types of produced enzymes and safety; and enzyme activity is considerable and is daily kept at 90,000-110,000, and the highest enzyme activity can reach about 120,000.

Description

technical field [0001] The invention relates to the field of enzyme preparations, in particular to the preparation of an acid protease-based enzyme preparation, its bacterial strain and its application. Background technique [0002] With the growing demand for poultry products, the importance of the breeding industry to people has increased year by year, and higher requirements have been put forward for the safety, efficiency and environmental protection of poultry breeding. "Food is the most important thing for poultry", and feed, as one of the key links in the breeding industry, is highly valued. In terms of the situation, the use of antibiotics has become more and more strictly controlled in recent years. China will enter the era of "banning antibiotics" in 2020, and the detection of prescribed antibiotics in feed additives is prohibited. The dual trends have directly promoted the upsurge in the quality optimization of feed production processes, additives and other produ...

Claims

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Application Information

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IPC IPC(8): C12N9/62C12N9/42C12N9/18C12N9/26C12N9/88C12N9/24C12N9/38C12N9/40C12N9/48C12N9/16C12N1/14A23K20/189C12R1/685
CPCC12N9/62C12N9/2437C12N9/88C12N9/18C12N9/248C12N9/2411C12N9/2488C12N9/2445C12N9/2408C12N9/2465C12N9/2471C12N9/485C12N9/16C12Y301/01073C12Y302/01022C12Y302/01023C12Y302/0102C12Y302/01021C12Y301/01011C12Y302/01015C12Y402/02002C12N1/14A23K20/189C12R2001/685C12N1/145Y02P60/87
Inventor 王云龙吴勃王天珍徐永雷王云祥
Owner HANGZHOU BIOCOM BIOLOGICAL TECH
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