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Xylanase production microbiological screening culture medium and culture method thereof

A technology of xylanase activity and microorganisms, applied in the field of microorganisms, can solve the problems of large workload, large experimental workload, large labor load, etc., and achieve the effect of being convenient to carry.

Inactive Publication Date: 2015-11-18
HAIKOU EXPERIMENTAL STATION CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Second, the screening method is too fine, so that the experimental workload of the entire screening process is too large, resulting in a large labor load and ultimately resulting in low efficiency
The use of the transparent circle method in the primary screening and the first re-screening is especially too fine, resulting in excessive workload in the whole process
In addition, in xylanase-related utilization factories, the preservation of strains generally adopts the conventional method of strain preservation, which can easily lead to the degradation of the enzyme activity of the strains.

Method used

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  • Xylanase production microbiological screening culture medium and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Preparation of microbial screening medium:

[0057] Commercially available carrots are cut into round slices, the thickness D3 of the carrot slices is 6-10mm, and the diameter D1=50-70mm. On the previously cut carrot slices, use a special machine to carve a "Z"-shaped groove. The size of the groove is similar to the shape of carrot slices, and the diameter D2figure 1 As mentioned above, the thickness D3 of the carrot slices is 6-10mm; the groove depth: 3mm, width: 1mm.

[0058] Pour the pasteurized nutrient solution into the groove of the carrot slices to make it evenly distributed in the groove; the nutrient solution is formed by mixing the tomato juice and the substrate, the tomato juice and the The volume ratio of substrate was 300:1.

[0059] Wherein, the tomatoes are ground through a homogenizer until there are no granular fragments, and the filtrate is filtered through a 20-mesh sieve, and the sieve holes are covered with 1 to 2 layers of toilet paper; the filtrat...

Embodiment 2

[0064] Rapid screening of xylanase-producing strains

[0065] Using the microbial screening medium in Example 1, after the carrot was cut into slices, a continuous groove was scored on it, and the mixed solution of tomato juice, sample solution and substrate was poured in the groove, and the volume ratio of the three was 300:100:1, so that the above mixture is evenly distributed in the groove, and cultured at room temperature for 2-4 days to obtain the microorganisms that form wounds on the surface of carrot slices in the microbial screening medium, and transfer them to the microbial screening culture every day after 2 days Based on the new microbial colonies, these microorganisms were further re-screened by the transparent circle method.

[0066] After further verification by the transparent circle method, the positive rate of primary screening microorganisms based on this method reaches 89%, while the conventional transparent circle method is used for primary screening. Afte...

Embodiment 3

[0071] Rapid Screening of Xylanase, Mannanase and Pectinase Complex Enzyme-producing Strains

[0072] Using the method of Example 2 to obtain the starting strains, combined with a large number of marine microorganisms preserved in the laboratory's previous work, a total of more than 10,000 starting strains were obtained, and 410 strains with xylanase activity were obtained. Add pectinase substrate, mannanase substrate and xylanase substrate respectively to LB medium, and add corresponding color indicator to make pectinase screening plate, mannanase screening plate and Xylanase Screening Plate. Spot the strains to be tested on pectinase, mannanase, and xylanase screening plates, culture at a constant temperature at a constant temperature, and observe the changes around the colonies. If the surroundings of the colonies become transparent, this is the hydrolysis circle. , it shows that the strain produces the enzyme with corresponding activity.

[0073] After several experiment...

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Abstract

The invention provides a xylanase production microbiological screening culture medium, a method for microorganism culture by virtue of the microbiological screening culture medium, application of the microbiological screening culture medium in preliminary screening of enzymatic activity strains as well as application of the microbiological screening culture medium in strain preservation and rejuvenation. The method disclosed by the invention is applicable to field operation; and collected samples can be processed on the same day, so as to avoid the trouble of taking the field samples back to a laboratory.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a xylanase-producing microorganism screening medium and a culture method thereof. Background technique [0002] The traditional method for screening xylanase enzyme-producing strains is to first isolate and culture microorganisms from nature, then use randomly obtained pure culture microorganisms as starting strains, inoculate on agar plates containing xylan substrates, and add certain The concentration of the pigment indicator, through the pigment, the enzyme-producing active strain will produce a corresponding transparent circle, so that the enzyme-producing active strain can be screened out. This method is generally called the transparent circle method. [0003] There are mainly two kinds of this disadvantage. First, it is not conducive to field work. Soil or other samples collected in the field must be brought back to the laboratory for isolation and culture. After obtaining a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04
Inventor 郭刚张凤娟杨晓琳陈乾郭素霞
Owner HAIKOU EXPERIMENTAL STATION CHINESE ACAD OF TROPICAL AGRI SCI