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Streptomyces polysaccharide degradation bacterium as well as culture method and application thereof

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 of various types and the like

Active Publication Date: 2017-02-22
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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  • Streptomyces polysaccharide degradation bacterium as well as culture method and application thereof
  • Streptomyces polysaccharide degradation bacterium as well as culture method and application thereof
  • Streptomyces polysaccharide degradation bacterium as well as culture method and application thereof

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 degradation bacterium as well as a culture method and application thereof. A streptomyces sp. CB16 strain is preserved in China General Microbiological Culture Collection Center on April 11, 2016 and the preservation number is CGMCC No.12351. The invention further relates to the culture method and the application of the strain. The strain provided by the invention is a multifunctional polysaccharide degradation bacterium; a compound type polysaccharide degradation enzyme preparation prepared from the strain can be used for degrading plant, seaweed, animal and microorganism polysaccharides and particularly has remarkable degradation activity on hyaluronic acid, chondroitin sulfate A, chondroitin sulfate C and chondroitin sulfate E from connective tissues of higher animals; and the streptomyces polysaccharide degradation bacterium has a potential application value and has functions which are completely different from functions of existing 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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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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