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Method for separating wuyiencin from fermentation liquor

A technology of Wuyiencin and fermentation liquid, which is applied in the field of bioengineering, can solve the problems of low purity and low extraction efficiency of Wuyiencin, and achieve high-efficiency extraction

Active Publication Date: 2018-10-16
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the problems existing in the prior art, the purpose of the present invention is to propose a method for efficiently extracting high-purity Wuyiencin in view of the problems of low extraction efficiency and low purity of Wuyiencin, and no toxic and harmful chemical organic substances are used in the whole process. The reagent has the characteristics of high extraction rate, cleanness, environmental protection, simplicity, and high efficiency. The extraction rate is greatly improved compared with previous processes, and the purity can reach more than 95%.

Method used

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  • Method for separating wuyiencin from fermentation liquor
  • Method for separating wuyiencin from fermentation liquor
  • Method for separating wuyiencin from fermentation liquor

Examples

Experimental program
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Effect test

Embodiment 1

[0064] 1. Fermentation and cultivation of Wuyiencin-producing strains

[0065] Use a scalpel to cut the Wuyiencin-producing strain - Streptomyces nonhygroscopicus Wuyi var. CK15 from the MS medium by 2 cm 2 Inoculate into Erlenmeyer flasks equipped with 200ml of fermentation medium, shake and culture at 28°C and 220rpm for 64h on a shaker, and ferment 30 flasks in total.

[0066] MS medium components are: mannitol 20g / L, soybean powder 20g / L, agar powder 20g / L.

[0067] The composition of the fermentation medium is: corn flour 20g / L, soybean flour 25g / L, glucose 25g / L, calcium carbonate 4g / L.

[0068] 2. Crude extraction of Wuyiencin

[0069] (1) Pretreatment of fermented liquid: Centrifuge 30 bottles of fermented liquid at room temperature (9500rpm, 15min), remove culture medium residue and bacterial cells therein, collect 4.8L fermented liquid altogether, determine by HPLC, titer is 1100ppm;

[0070] (2) Add oxalic acid to the fermentation broth, adjust the pH to 2.0-2.5,...

Embodiment 2

[0086] 1. Fermentation and cultivation of Wuyiencin-producing strains

[0087](1) Strain culture: inoculate CK15 onto 10 MS plates with a diameter of 9 cm, cultivate at 28°C for 7 days, and then use 50 ml of sterile water to elute the spores into a 200 ml Erlenmeyer flask;

[0088] The composition of MS medium is the same as that in Example 1.

[0089] (2) Seed liquid culture: inoculate the above spore liquid into a 50L fermenter with 30L of culture medium, at a temperature of 28°C, a speed of 200rpm, an air flow ratio of 1:0.5, and a tank pressure of 0.1kg / cm 2 Cultivate 24h under the condition of obtaining seed liquid;

[0090] The medium components are: glucose 10g / L, peptone 5g / L, yeast powder 3g / L, sucrose 260g / L.

[0091] (3) Fermentation tank culture: Transfer the vigorously growing primary seed liquid into a 500L fermenter tank with 300L fermentation medium inside, at a temperature of 28°C, a speed of 200rpm, an air flow ratio of 1:0.55, and a tank pressure of 0.12 ...

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Abstract

The invention relates to a method for separating wuyiencin from fermentation liquor. The method comprises the following steps: 1) removing culture medium solid residues and thalli in the fermentationliquor which contains the wuyiencin to obtain supernate; 2) acquiring impurities in the supernate obtained from step 1) by precipitating through oxalic acid, and then centrifuging to remove precipitates and impurities to obtain a solution; 3) treating the solution obtained from step 2) by macroporous resin, and collecting effluent liquid; 4) carrying out gel chromatography treatment on the effluent liquid collected from step 3), and separately collecting the effluent liquid which flowing out in different time periods; 5) detecting the effluent liquid collected from step 4) by high performanceliquid chromatography, freeze-drying the effluent liquid which has the wuyiencin but does not have miscellaneous peaks to obtain the crude product of the wuyiencin; and 6) preparing and refining the crude product freeze-dried in step 5) by using a liquid phase preparation system or a liquid phase semi-preparation system to obtain high-purity wuyiencin. The method for separating the wuyiencin fromthe fermentation liquor has the advantages of high extracting rate, high wuyiencin extracting purity and the like.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for efficiently separating and purifying Wuyiencin from fermentation broth. Background technique [0002] Wuyimectin is a low-toxicity, high-efficiency, broad-spectrum, and environmentally friendly new type of agricultural antibiotic with completely independent intellectual property rights in my country. It is produced by fermentation of Streptomyces ahygroscopicus varietal Wuyi. It is a water-soluble nucleoside antibiotic with a new structure. , and its molecular structure is: [0003] [0004] Wuyiencin was independently developed by the Plant Protection Institute of the Chinese Academy of Agricultural Sciences. It has obvious control effects on fungal diseases of various crops such as vegetables and fruit trees. It has been widely used in agricultural production and created considerable economic, social and social benefits. Ecological Benefits. ...

Claims

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Application Information

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IPC IPC(8): C07H19/067C07H1/06
CPCC07H1/06C07H19/067
Inventor 施李鸣葛蓓孛张克诚刘炳花赵文珺麻金金韦秋合
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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