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Flat-plate sieving method of aesculin for high-yield beta-glucosidase producing strain

A technology of glucosidase and escin, applied in biochemical equipment and methods, microbial determination/inspection, enzymes, etc., can solve the problems of expensive reagents, cumbersome methods, etc., and achieve reliable results, simple operation, and obvious price. The effect of advantage

Inactive Publication Date: 2008-05-14
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0003] Aiming at the problems that the reagents used in the current β-glucosidase screening method are relatively expensive and the method is cumbersome, and the research status of a fast and simple screening method is urgently needed in scientific research work, the problem to be solved in the present invention is to provide a method for using aescin Method for plate screening of β-glucosidase producing bacteria

Method used

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  • Flat-plate sieving method of aesculin for high-yield beta-glucosidase producing strain
  • Flat-plate sieving method of aesculin for high-yield beta-glucosidase producing strain
  • Flat-plate sieving method of aesculin for high-yield beta-glucosidase producing strain

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Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1: screening obtains Aspergillus Asp-1

[0025] Collect 10 soil samples from the rural wheat straw piles and dead branches and leaves in the woods, and then accurately weigh 0.5g of each soil sample and dissolve it in 25ml distilled water. 30°C, 180r / min shaker for enrichment culture for two days, and then diluted 10 1 , 10 2 , 10 3 , 10 4 , 10 5 , 10 6 times, and then draw 0.1mL of the diluted solution evenly spread on the plate added with the basic inorganic salt medium with a mass volume percentage of 0.1% aescin, culture in a 30°C incubator for 36 hours, and carry out preliminary screening. On a plate with a suitable dilution factor and uniform growth of colonies, the color of the culture medium around the colony on the back of the plate becomes black as an index to screen and pick a colony with dark color on the back of the plate, named Aspergillus Asp-1 (for the results of plate culture, see figure 1). The colony Asp-1 was inoculated in the enzym...

Embodiment 2

[0034] Embodiment 2: screening obtains Aspergillus Asp-2

[0035] Collect 8 soil samples from the evergreen holly trees in the campus, then accurately weigh 0.5g of each soil sample and dissolve it in 25ml of distilled water. Min shaker for enrichment culture for two days, and then they were diluted 10 1 , 10 2 , 10 3 , 10 4 , 10 5 , 10 6 times, and then draw 0.3mL of the diluted solution evenly spread on the plate added with the basic inorganic salt medium with a mass volume percentage of 0.3% aescin, and culture it in a 20°C incubator for 72 hours, and carry out preliminary screening. On a plate with a suitable dilution factor and uniform growth of colonies, the color of the culture medium around the colony on the back of the plate becomes black as an index to screen and pick a colony with dark color on the back of the plate, named Aspergillus Asp-2 (see the results of plate culture) figure 2). The colony Asp-2 was inoculated in the enzyme-producing medium, cultured o...

Embodiment 3

[0044] Embodiment 3: screening obtains Aspergillus Asp-3

[0045] Collect 12 soil samples from the shady forest of Mount Tai, then accurately weigh 0.5g of each soil sample and dissolve in 25ml of distilled water, shake and mix well, absorb 1ml each into the enrichment medium, place at 50°C, shake at 220r / min The enrichment culture was carried out on the bed for 20 hours, and then it was diluted by 10 1 , 10 2 , 10 3 , 10 4 , 10 5 , 10 6 times, and then draw 0.2mL of the dilution evenly spread on the plate added with the basic inorganic salt medium with a mass volume percentage of 3% aescin, and cultivate it in a 50°C incubator for 12 hours, and carry out preliminary screening. On a plate with a suitable dilution factor and uniform growth of colonies, the color of the culture medium around the colony on the back of the plate becomes black as an index to screen and pick a colony with dark color on the back of the plate, named Aspergillus Asp-3 (for the results of plate cul...

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Abstract

The invention discloses a flat plate screening method for high yielding Beta-glucosidase production bacteria by using aesculin. The method comprises the steps of: 1. earth sample collection; 2. enrichment culture; 3. flat plate preparation of the aesculin; 4. flat plate primary screening aesculin; 5. separation and purification; 6. secondary screen by a shaking battle, etc. The invention can fast screen active strain possessing the Beta-glucosidase from samples which contain rich fibrin resources; compared with the method which is reported in published literature which screens Beta-glucosidase production bacteria by using a cellobiose flat plate method, the invention has the remarkable advantages at price; under the condition that obtains 100 strains by screening, the cost of the invention by using the aesculin is only one tenth of the cellobiose flat plate method; in addition, the invention has the advantages of simple operation, reliable result, etc.

Description

technical field [0001] The invention relates to plate screening of β-glucosidase-producing bacteria, in particular to a method for using aescin for plate screening of high-yielding β-glucosidase-producing bacteria. Background technique [0002] In the process of hydrolyzing natural cellulose into glucose, it must rely on the synergistic action of three enzymes, endo-cellulase, exo-cellulase and β-glucosidase. Among them, β-glucosidase is the rate-limiting enzyme in this process, and the level of its enzyme activity directly affects the level of the overall enzyme activity of the cellulase system. Most of the most widely used cellulase producing strains are excellent mutant strains of Trichoderma reesei. Although these strains can produce endo- and exo-cellulase with high activity, due to the low activity of β-glucosidase, in the process of hydrolyzing cellulose with cellulase, the accumulation of cellobiose, cellobiose Sugar, in turn, forms a strong feedback inhibition of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12N1/14C12N1/21C12N9/42
Inventor 宋欣曲音波袁晓华
Owner SHANDONG UNIV
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