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Phylum porifera symbiosis streptomycete, method for producing staurosporine by fermenting phylum porifera symbiosis streptomycete and application of phylum porifera symbiosis streptomycete

A staurosporine-sponge symbiosis technology, applied in microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of low yield, inability to meet industrial applications, and low staurosporine yield, etc. achieve high yield

Inactive Publication Date: 2018-01-19
HAINAN UNIVERSITY
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  • Claims
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Problems solved by technology

[0004] Streptomyces staurosporine production in the current prior art is not high, and the fermentation cost is high, mainly used in anti-cancer research
For example, it is recorded in the "Combining Biological and Chemical Methods to Increase the Production of Staurosporine of Actinomycetes YB-24 Strain in Marine Habitats" published on the fifth issue of volume 40 of Chinese Journal of Antibiotics in May 2015, after ultraviolet light and lithium chloride The staurosporine yield of the high-yield strain obtained after compound mutagenesis was 3.833 mg / L, which was still low and could not meet industrial applications

Method used

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  • Phylum porifera symbiosis streptomycete, method for producing staurosporine by fermenting phylum porifera symbiosis streptomycete and application of phylum porifera symbiosis streptomycete
  • Phylum porifera symbiosis streptomycete, method for producing staurosporine by fermenting phylum porifera symbiosis streptomycete and application of phylum porifera symbiosis streptomycete
  • Phylum porifera symbiosis streptomycete, method for producing staurosporine by fermenting phylum porifera symbiosis streptomycete and application of phylum porifera symbiosis streptomycete

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Isolation and Identification of Streptomyces (Streptomyces) SP6206

[0053] 1. Isolation of Streptomyces sp6206

[0054] The composition of strain isolation medium is as follows: soluble starch 10g, casein 0.3g, potassium nitrate 2g, magnesium sulfate heptahydrate 0.05g, sodium chloride 2g, dipotassium hydrogen phosphate 2g, calcium carbonate 0.02g, ferrous sulfate heptahydrate 0.01 g, potassium dichromate 0.1g, distilled water 500ml, aged sea water 500ml, agar 20g, pH 7.3.

[0055] Weigh 1g of sponge sample (collected from the sea area of ​​Sanya, Hainan), grind it with 10ml of sterile seawater, then dilute it with sterile seawater 10 times, 100 times, 1000 times, absorb 100μl dilution respectively, and spread it on the separation medium plate , and cultured upside down in a constant temperature incubator at 28°C for 1 to 4 weeks. According to the morphological characteristics of the colonies on the plate, the actinomycete colonies on the plate were picked ...

Embodiment 2

[0058] Embodiment 2 uses Streptomyces sp6206 to produce staurosporine

[0059] The preparation method of the seed medium is as follows (pH value 7.5): 4g of yeast extract powder, 10g of malt extract powder, 4g of glucose, 500mL of aged sea water, and the remaining tap water to the volume of 1L. Sterilize at 121°C for 20 minutes.

[0060] The preparation method of the fermentation medium is as follows (pH value 9.0): 31.3g of soluble starch, 15g of soybean powder, 2g of yeast extract powder, 4g of calcium carbonate, 12.5g of HP-20 macroporous resin, 500ml of seawater, and the remaining tap water To IL. Sterilize at 121°C for 20 minutes.

[0061] 1. Strain fermentation

[0062] 1. Seed culture: Inoculate 100 ml of seed culture medium with Streptomyces sp6206. Cultivate at 28°C and 180 rpm for 96 hours to obtain a seed liquid.

[0063] 2. Fermentation culture: Fill 200ml fermentation medium in a 500ml Erlenmeyer flask. Inoculate 2ml of seed liquid in the culture medium, cul...

Embodiment 3

[0085]Embodiment 3 application Streptomyces SP6206 fermentation production staurosporine

[0086] The preparation method of the seed medium is as follows (pH7.5): 4g of yeast extract powder, 10g of malt extract powder, 4g of glucose, 500mL of aged sea water, and the remaining tap water to the volume of 1L. Sterilize at 121°C for 20 minutes.

[0087] The preparation method of the fermentation medium is as follows (pH9.0): 25.4g of soluble starch, 18g of soybean powder, 1.5g of yeast extract powder, 3g of calcium carbonate, 20.5g of HP-20 macroporous resin, 450ml of seawater, and the remaining tap water To IL. Sterilize at 121°C for 20 minutes.

[0088] 1. Strain fermentation

[0089] 1. Seed culture: Inoculate 100 ml of seed culture medium with Streptomyces sp6206. Cultivate at 28°C and 180 rpm for 96 hours to obtain a seed solution.

[0090] 2. Fermentation culture: Fill 200ml fermentation medium in a 500ml Erlenmeyer flask. Inoculate 3 ml of seed solution in the culture...

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Abstract

The invention relates to phylum porifera symbiosis streptomycete SP6206, belonging to the field of phylum porifera symbiosis streptomycete fermentation. The preservation number of the phylum poriferasymbiosis streptomycete SP6206 is CCTCCM2017451. The invention also discloses a method for producing staurosporine by fermenting the phylum porifera symbiosis streptomycete SP6206, which comprises thefollowing steps: (1) culturing a seed solution of the SP6206; (2) inoculating the cultured seed solution of the SP6206 to a fermenting culture substrate, and carrying out fermented culture, thereby obtaining a fermentation solution; and (3) separating staurosporine from the fermentation solution. According to the phylum porifera symbiosis streptomycete SP6206, the highest yield of the staurosporine reaches 320mg / L and is greatly increased compared with that of the common wild streptomycete staurosporine in the prior art. The staurosporine can be used for preventing and treating the colletotrichum gloeosporioides of Chinese yams, the minimal inhibitory concentration of the staurosporine for preventing and treating the colletotrichum gloeosporioides of the Chinese yams is 8 microg / ml, and the minimal inhibitory concentration of the staurosporine is 1 / 4 that of the positive drug cycloheximide.

Description

technical field [0001] The invention relates to the technical field of fermentation of Streptomyces spongiosa, in particular to Streptomyces spongiosa and its fermentation production method and application of staurosporine. Background technique [0002] Sponges are invertebrate marine animals belonging to the phylum Porezoa. There are many types of rich and novel marine symbiotic microorganisms living inside and outside their tissues. The alkaloids, lactones, quinones, ketones, polyethers, terpenes produced by these symbiotic microorganisms A variety of active secondary metabolites such as alkenes and steroids are becoming an important source for the screening of marine drug lead compounds. [0003] Staurosporine is the first indolecarbazole alkaloid isolated from the fermentation product of Streptomyces staurosporeus AM-2282 in 1977. In 1986, it was discovered that this alkaloid has anti-tumor effect, and belongs to a class of non-selective protein kinase inhibitors with g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/18C07D498/22A01N43/90A01P3/00C12R1/465
Inventor 黄小龙黄东益周双清许云吴文嫱刘柱郑继平陶琳
Owner HAINAN UNIVERSITY
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