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Agrobacterium radiobacter mutant strain and method for preparing beta-1, 3 glucan as fish and shrimp disease-resistant factor feed additive through utilizing agrobacterium radiobacter mutant strain

An Agrobacterium and radial technology, applied in the field of bioengineering, can solve problems such as unsuitable for farmers, high product price, long process flow, etc., and achieve the effect of enhancing the immunity of prawns, low cost, and short process process

Active Publication Date: 2012-12-19
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of this method are: the process is long, the equipment is various, the investment is large, and there is a large amount of wastewater discharge, the product is expensive, and it is not suitable for farmers to use

Method used

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  • Agrobacterium radiobacter mutant strain and method for preparing beta-1, 3 glucan as fish and shrimp disease-resistant factor feed additive through utilizing agrobacterium radiobacter mutant strain
  • Agrobacterium radiobacter mutant strain and method for preparing beta-1, 3 glucan as fish and shrimp disease-resistant factor feed additive through utilizing agrobacterium radiobacter mutant strain
  • Agrobacterium radiobacter mutant strain and method for preparing beta-1, 3 glucan as fish and shrimp disease-resistant factor feed additive through utilizing agrobacterium radiobacter mutant strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. Activate the Agrobacterium radiata purchased from the China Typical Culture Collection Center with the preservation number As.1.150 according to the instructions provided by the Culture Collection Center, transfer it to the slant medium, and store it for future use .

[0052] 2. The medium formula used in the whole process of the experiment:

[0053] A: Incline medium:

[0054] 0.3% beef extract, 0.5% peptone, 0.5% sodium chloride, 0.1%-0.15% corn steep liquor hydrolyzate, 2% agar, adjusted to pH 7.0-7.8, and routinely sterilized.

[0055] B: Plate screening medium:

[0056] 1% glucose, 0.5% yeast extract, 0.001%-0.008% aniline blue dye, 2% agar, adjust pH to 7.0-7.8, routine sterilization

[0057] C: basic fermentation medium:

[0058] 4% sucrose, 0.1% diammonium hydrogen phosphate, 0.1% potassium dihydrogen phosphate, 0.05% magnesium sulfate heptahydrate, 0.005% ferrous sulfate heptahydrate, 0.1%—0.15% corn steep liquor hydrolyzate, 0.002% cobalt chloride, 0.00...

Embodiment 2

[0106] Steps one to six are the same as in Example 1.

[0107] 7. Preparation of β-1,3 glucan fish and shrimp disease resistance factor feed additive

[0108] 1. Activation of strains

[0109] The excellent mutant strain L-15 stored in a refrigerator at 4°C was re-inoculated on a slant medium and cultured at 30°C for 48 hours.

[0110] 2. liquid seed culture

[0111] Take the activated strain L-15, pick a loop of bacteria with an inoculation loop, inoculate it into a 250mL Erlenmeyer flask containing 50mL basic fermentation medium, place it on a shaker at 35°C, and shake it at 160r / min for 24 hours.

[0112] 3. Primary expansion culture of bacteria

[0113] Take the liquid strains that have been cultured for 24 hours, add 10% of the inoculum to a 500mL Erlenmeyer flask containing 200mL of basic fermentation medium, place it at 35°C, and shake at 160r / min for 24 hours.

[0114] 4. Take the cultivated first-level seed culture solution, and use aseptic operation according ...

Embodiment 3

[0119] Steps one to six are the same as in Example 1.

[0120] 7. Preparation of β-1,3 glucan fish and shrimp disease resistance factor feed additive

[0121] 1. Activation of strains

[0122] The excellent mutant strain L-15 stored in a refrigerator at 4°C was re-inoculated on a slant medium and cultured at 32°C for 30 hours.

[0123] 2. liquid seed culture

[0124] Take the activated strain L-15, pick a ring of bacteria with an inoculation loop, inoculate it into a 250mL Erlenmeyer flask with 50mL basic fermentation medium, place it on a shaker at 32°C, and culture it with shaking at 180r / min for 24 hours .

[0125] 3. Primary expansion culture of bacteria

[0126] Take the liquid strains that have been cultured for 24 hours, add 8% of the inoculum to a 500mL Erlenmeyer flask containing 200mL of basic fermentation medium, place it at 32°C, and shake at 180r / min for 30 hours.

[0127] 4. Take the cultivated first-level seed culture liquid, and use aseptic operation ac...

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Abstract

The invention discloses an agrobacterium radiobacter mutant strain which is classified and named as an agrobacterium radiobacter mutant strain L-15, and has the preservation number of CCTCC (China center for type culture collection) NO: M2012 294. A method for preparing a beta-1, 3 glucan as a fish and shrimp disease-resistant factor feed additive through utilizing the agrobacterium radiobacter mutant strain L-15 comprises the steps that the agrobacterium radiobacter mutant strain L-15 is inoculated in an inclined culture medium and a basic fermentation culture medium to obtain a secondary seed culture liquid; and the secondary seed culture liquid is added into a 50 L seed fermentation tank filled with 35 L basic fermentation culture medium for culturing for 24-48 hours, the obtained seed liquid is filled in a 500L fermentation tank filled with 350L fermentation culture medium for culturing for 90-100 hours, so the fermentation liquor with the beta-1, 3 glucan content being above 4% is obtained. The method has the advantages of high production efficiency, low cost, short technological process, no fermentation liquor sewage discharge, environmental friendliness and the like.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a mutant strain of Agrobacterium radiata and a method for preparing β-1,3 glucan as a feed additive for fish and shrimp disease resistance factors. Background technique [0002] β-1,3 glucans are ubiquitously present in the cell walls of various bacteria, fungi, plants and yeasts. Scientists from Finland and other countries in northern Europe in the last century have experimentally proved that β-1,3 glucan can improve the immunity of fish and shrimp against diseases. The principle is that there is a highly specific receptor on the macrophages of fish and shrimp. This kind of receptor can highly transfer and bind the special molecular structure of β-1,3 glucan, thereby activating macrophages and enhancing their ability to phagocytose various germs and viruses. In addition, β-1,3 glucan can also react with a specific protein in the blood of crustaceans (such as shrimp, et...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/01C12N13/00C12P19/08A23K1/16A23K1/18C12R1/01
Inventor 周河治杨辉周延华梁海秋
Owner GUANGXI UNIV
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