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Method for obtaining Rosellinia sp.fungi imperfecti spore-free mycete capable of producing PF1022

A technology of PF1022 and half-knowledge bacteria, which is applied in the field of obtaining half-knowledge fungus aseptic molds for the production of PF1022, can solve the problems of low yield of PF1022A, selection of unfavorable strains, and small difference in colony morphology, etc., to achieve industrial production and yield High, yield-enhancing effects

Inactive Publication Date: 2017-09-29
SHANGHAI INST OF PHARMA IND CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, this strain does not produce sexual spores or asexual spores, and the morphological differences of colonies on solid plates are small, which is not conducive to the selection of strains based on phenotype
At present, there are relatively few reports on the production of PF1022A by fermentation of PF1022, and the yield of PF1022A is relatively low.

Method used

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  • Method for obtaining Rosellinia sp.fungi imperfecti spore-free mycete capable of producing PF1022
  • Method for obtaining Rosellinia sp.fungi imperfecti spore-free mycete capable of producing PF1022
  • Method for obtaining Rosellinia sp.fungi imperfecti spore-free mycete capable of producing PF1022

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The composition of the hyperosmotic plate solid medium is: 20% potato, 2% glucose, 1.8% agar and KCl with a concentration of 0.7 mol / L, and the balance is water, and the percentages are mass volume percentages.

[0031] The hyperosmotic plate solid medium is prepared by the following method: cut 200g of potatoes into small pieces and put them in a pot, add water and heat to boiling, keep for 20min, filter with 8 layers of gauze, add water to 1000ml, add 20g of glucose , 18g agar and 0.7mol KCl.

[0032] Except that the composition of the slant medium does not contain KCl, other components and preparation methods are the same as those of the hyperosmotic plate solid medium.

[0033] The composition of the solid culture medium of the comparison plate is: 20% potato, 2% glucose and 1.8% agar, and the balance is water, and the percentage is mass volume percentage.

[0034] (1), after grinding Rosellinia.sp.PF1022 preserved in glycerol in a homogenizer, dilute to 10 -3 ~10...

Embodiment 2

[0041] The composition of hyperosmotic plate solid medium is: 20% potato, 2% glucose, 1.8% agar and concentration are respectively 0.1mol / L, 0.3mol / L, 0.5mol / L, 0.7mol / L, 0.9mol / L or The KCl of 1mol / L, balance is water, and described percentage is mass volume percentage.

[0042] Cut 200g of potatoes into small pieces and put them in a pot, add water, heat to boiling, keep for 30min, filter with 8 layers of gauze, add water to 1000ml, add 2g of glucose, 18g of agar and KCl.

[0043] The composition of the slant medium is: 20% potato, 2% glucose and 1.8% agar, and the balance is water, and the percentage is mass volume percentage.

[0044] The preparation method of the slant medium is the same as the preparation method of the hypertonic plate solid medium.

[0045] (1), after grinding Rosellinia.sp.PF1022 preserved in glycerol in a homogenizer, dilute to 10 -3 ~10 -4 1 / ml to obtain the dilution, take 0.1ml of the dilution and spread it evenly on the solid medium of the hyper...

Embodiment 3

[0054] The composition of hyperosmotic plate solid medium is: 20% potato, 2% glucose, 1.8% agar and concentration are respectively 0.1mol / L, 0.3mol / L, 0.5mol / L, 0.7mol / L, 0.9mol / L or 1mol / L MgSO 4 .7H 2 O, the balance is water, and the percentage is mass volume percentage.

[0055] The composition of the slant medium is: 20% potato, 2% glucose and 1.8% agar, and the balance is water, and the percentage is mass volume percentage.

[0056] All the other conditions are completely consistent with Example 2. The results are shown in Table 3, and the data in Table 3 illustrate that different concentrations of MgSO 4 .7H 2 O can affect the fermentative unit of PF1022A, with MgSO 4 .7H 2 With the increase of O concentration, the positive mutation rate of the obtained strain also showed a corresponding increase. Wherein when MgSO 4 .7H 2 When the concentration of O was 0.5mol / L or 0.7mol / L, the rate of positive mutation increased significantly. Example 3 The results produced ...

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Abstract

The invention discloses a method for obtaining fungi imperfecti spore-free mycete Rosellinia sp. capable of producing PF1022. The method comprises the following steps: 1) culturing wild Rosellinia sp.fungi imperfecti spore-free mycete strains in a flat-plate solid culture medium containing 0.1mol / L to 1.2mol / L of a hypertonic solution; selecting single colonies with the colony diameter of less than 5mm and a round shape; 2) inoculating the single colonies into a fermentation culture medium and culturing; detecting the content of PF1022A in a fermentation product; selecting strains with the PF1022A content which is higher than that of starting strains. By adopting the method disclosed by the invention, the fungi imperfecti spore-free mycete strains Rosellinia sp. Strain PF1022 with the PF1022A yield which is improved by at least 1.2 times can be obtained; furthermore, the method has good repeatability and good reliability and is widely applied to a process of producing the PF1022 by utilizing the Rosellinia sp.fungi imperfecti spore-free mycete and industrial application of the PF1022A is facilitated.

Description

technical field [0001] The invention belongs to the technical field of strain breeding, and in particular relates to a method for obtaining a deuteromorphic fungus asporosa that produces PF1022. Background technique [0002] PF1022A is an N-methyl-CODP (cyclic depsipeptide) compound produced by the fungus Aspenia sp. Rosellinia sp. PF1022. PF1022A is a kind of anthelmintic drug with low toxicity to animals, wide control spectrum and good effect. After macrolide anthelmintic drugs, it is the most potential class of anthelmintic drugs for livestock and pets. [0003] US patent document US09901393A1 (published on April 18, 2002) discloses a chemical method for converting PF1022A to produce its derivative emodepside. Profender, a combination of emodepside and praziquantel, is currently on the market for the control of nematodes and tapeworms in the gastrointestinal tract of cats or dogs. Subsequently, Procox, a combination of emodepside and toltrazuril, has also been launched....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12R1/645
CPCC12N1/14
Inventor 陈少欣刘瑞
Owner SHANGHAI INST OF PHARMA IND CO LTD
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