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A Streptomyces polysaccharide-degrading bacterium and its cultivation method and application

A culturing method and Streptomyces technology are applied in the field of Streptomyces polysaccharide degrading bacteria and their cultivation fields, and can solve the problems of rare single Streptomyces polysaccharide degrading enzymes and other types of polysaccharide degrading enzymes

Active Publication Date: 2019-05-28
WUTONG AROMA CHEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies on polysaccharide-degrading enzymes produced by Streptomyces mainly focus on single polysaccharide-degrading enzymes produced by the strain, such as chitinase, xylanase, and cellulase. [11-13] , it is rare to report that a single Streptomyces can produce multiple types of polysaccharide degrading enzymes

Method used

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  • A Streptomyces polysaccharide-degrading bacterium and its cultivation method and application
  • A Streptomyces polysaccharide-degrading bacterium and its cultivation method and application
  • A Streptomyces polysaccharide-degrading bacterium and its cultivation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the separation and purification of soil microorganisms in the root system of Fritillaria sichuan

[0045] Take the root soil leaching solution of Fritillaria sichuanensis, add 1 mL of the supernatant to 9 mL of sterile water, and dilute to a concentration of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 5 concentration gradients. Thoroughly mix the diluted bacterial suspension with Gaoshi No. 1 medium at about 50°C, make two parallels for each concentration, and culture at 28°C for 7 days. Pick out the actinomycetes colony with a more typical form, and then separate and purify it by streaking on the plate for 3 times, then pick a single colony in TSB liquid medium, cultivate it for 3 days at 28°C and 200 rpm, take 1.6ml of the culture and add it to 400μl glycerol, mix well and store in -80℃ refrigerator for a long time.

[0046] The above-mentioned TSB liquid medium has the following components per liter:

[0047] Tryptone 17g, plant peptone 3g, sodium chloride...

Embodiment 2

[0050] Embodiment 2, the screening of polysaccharide degrading bacteria

[0051] The strains obtained in Example 1 were inoculated on the sole carbon source liquid medium, cultured at 200 rpm and 30°C for 72 hours, and the turbidity of the bacterial liquid was observed. At the same time, the culture supernatant was taken for carbazole reaction to detect the carbon source. Consumption. The enzyme-producing strains were selected according to the above two indicators. The strains that were turbid in each unique carbon source medium and had a small carbazole reaction value were picked and cultivated on TSB solid medium, and the strains were recorded as CB16.

[0052] The preparation method of the above-mentioned sole carbon source medium is:

[0053] Add polysaccharide substrates to the ionic medium to a final concentration of 0.10% (w / v);

[0054] The above polysaccharide substrate is selected from: cellulose, carboxymethyl cellulose, xylan, mannan, chitin, chitosan, starch, p...

Embodiment 3

[0060] Example 3, Molecular Identification of Streptomyces sp. CB16 Strain Based on 16S rRNA Gene

[0061] Genomic DNA of the bacterial strain was prepared using the Bacterial Genome Extraction Kit (Tiangen), and the genomic DNA of the bacterial strain was used as a template to amplify using the universal primers of the bacterial 16S rRNA gene. After verification, it was connected to pEASY-Blunt Simple Cloning Vector and transformed into E.coli DH5α. After ampicillin resistance screening, positive clones were obtained. The 16S rRNA gene sequencing was completed by Sangon Bioengineering (Shanghai) Co., Ltd. The sequence was compared with the 16S rRNA gene sequence of standard strains collected by the National Center for Biological Information (NCBI), and a phylogenetic tree was constructed using MEGA6.0.

[0062] The above-mentioned general primers for the amplification of the strain 16S rRNA gene are:

[0063] The forward primer is 27f: 5'-AGAGTTTGATCCTG GCT CAG-3';

[0064...

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Abstract

The invention relates to a Streptomyces polysaccharide-degrading bacterium and its cultivation method and application. A strain of Streptomyces sp. CB16 was deposited on April 11, 2016 in the General Microbiology Center of China Committee for the Collection of Microbial Cultures, and the preservation number is CGMCC No.12351. The invention also relates to the culture method and application of the bacterial strain. The strain described in the present invention is a multipotent polysaccharide-degrading bacterium, and the compound polysaccharide-degrading enzyme preparation prepared by the strain can degrade polysaccharides of plants, seaweeds, animals and microorganisms, especially for hyaluronic acid, Chondroitin sulfate A, chondroitin sulfate C and chondroitin sulfate E have remarkable degradative activities and have potential application value, which are totally different from the known functions of Streptomyces sp.

Description

technical field [0001] The invention relates to a Streptomyces polysaccharide-degrading bacterium, a culture method and application thereof, and belongs to the field of biotechnology. Background technique [0002] Polysaccharides are polymers composed of repeating single sugar units or complex sugar units linked by glycosidic bonds, including linear molecules, branched chain molecules, or network polymers. There are many types and large quantities of polysaccharides, which widely exist in nature, such as: cellulose, xylan, mannan, etc. in higher plants, agar, alginate, and carrageenan in seaweed, and chitin in crustaceans quality, as well as peptidoglycan and xanthan gum from microbial sources. These polysaccharides are not only the structural substances that make up organisms, but also the medium for storing energy, and are one of the important biological resources. [1] . Moreover, polysaccharides and their degradation products have a variety of important physiological a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/24C12P19/14C12P19/12C12R1/465
CPCC12N1/20C12N9/24C12P19/12C12P19/14C12N1/205C12R2001/465
Inventor 韩文君古静燕刘会会李新卫洁
Owner WUTONG AROMA CHEM CO LTD
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