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A kind of separation and extraction method of Streptomyces fermentation metabolite neoaumycin

A technology for neoorthomycin and metabolites, which is applied in the field of separation and extraction of actinomycete fermentation metabolite neoorthomycin, can solve the problem of lack of downstream technology of fermentation engineering for extracting neoorthomycin, large activity loss, cumbersome steps, etc. question

Inactive Publication Date: 2016-08-31
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This extraction technology has cumbersome steps, consumes a large amount of organic solvents, has a long extraction cycle, and has a large loss of activity, so it is not suitable for the preparation of a large amount of neoaumycin
Invention patent ZL201110261822.1 discloses an extraction technology method in which the silica gel chromatography ternary elution system is changed to a binary elution system, and the steps of reverse phase silica gel chromatography separation and high performance liquid chromatography purification are removed, which consumes organic solvents less, etc., but the regeneration and use times of the silica gel column limit the scale-up production to a certain extent
At present, there is a lack of fermentation engineering downstream technology for extracting neoauthin from fermentation broth

Method used

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  • A kind of separation and extraction method of Streptomyces fermentation metabolite neoaumycin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The fermentation medium of Streptomyces knowlesi XiAo-3 (glycerin 1.0%, sucrose 7.0%, ammonium sulfate 0.1%, glucose 8.0%, peptone 2.0%, yeast powder 1.0%, magnesium sulfate 0.5%, potassium dihydrogen phosphate 1.0% , sodium chloride 0.5%, calcium chloride 0.5%) were sterilized at 121°C for 30 minutes, inoculated with Streptomyces noursii XiAo-3 seed solution after cooling, and fermented with aeration and stirring for 4 days at a temperature of 28°C-30°C. After the fermentation is over. First, heat the fermented liquid to 60°C and keep it for 30 minutes to kill the living bacteria of Streptomyces in the fermented liquid, then reduce the temperature of the fermented liquid to below 30°C, add 2.0% diatomaceous earth filter aid to sterilize In the final fermentation broth, stir evenly, let stand for 20 minutes, filter with a 600-mesh filter cloth, and collect the filtrate.

[0027] The filter cloth filtrate is adsorbed and treated with macroporous adsorbent resin. The ma...

Embodiment 2

[0032]The fermentation medium of Streptomyces knowlesi XiAo-3 (glycerin 1.0%, sucrose 7.0%, ammonium sulfate 0.1%, glucose 8.0%, peptone 2.0%, yeast powder 1.0%, magnesium sulfate 0.5%, potassium dihydrogen phosphate 1.0% , sodium chloride 0.5%, calcium chloride 0.5%) were sterilized at 121°C for 30 minutes, inoculated with Streptomyces noursii XiAo-3 seed solution after cooling, and fermented with aeration and stirring for 4 days at a temperature of 28°C-30°C. After the fermentation is over. First, heat the fermentation broth to 60°C and keep it for 30 minutes to kill the living bacteria of Streptomyces noursii in the fermentation broth, then lower the temperature of the fermentation broth to below 30°C, and filter the above fermentation broth with a ceramic microfiltration membrane , remove medium solids and bacterial impurities in the fermentation broth, the separation pore size is 0.2 μm, the filtration pressure is 0.1 MPa, and the filtrate is collected.

[0033] The micr...

Embodiment 3

[0038] The method for removing medium solids and thalline impurities in the fermentation broth is the same as in Example 2. Treatment The filtrate is treated with a macroporous adsorption resin. The macroporous adsorption resin used is a mixture of D-101 nonpolar and polar macroporous adsorption resin in DM-301 to obtain the adsorption resin treatment liquid;

[0039] Cationic resin was used to enrich and separate neoauthin in the treatment solution of adsorption resin. What is used in the cation exchange process is 7320 sulfonic acid polystyrene gel type strongly acidic cation exchange resin. The adsorption resin treatment solution flows through the ion exchange column equipped with 7320 cation resin at an even speed, and the flow rate of the adsorption resin treatment solution is controlled so that the contact time between the adsorption resin treatment solution and the resin is 150 minutes.

[0040] The saturated resin was analyzed with 2% ammonia water, and the flow rate...

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Abstract

The invention discloses a separation and extraction method of streptomycete fermentation metabolite xinaomycin, which comprises the following steps: heating fermentation liquor to 60 DEG C to kill live streptomycete in the fermentation liquor, removing culture medium solids and thallus impurities in the xinaomycin fermentation liquor through a cloth filter or ceramic microfiltration membrane, adsorbing the solid-free fermentation liquor by a macroporous adsorbent resin, adsorbing with a cation exchange resin to enrich xinaomycin in the macroporous adsorbent resin effluent liquid, resolving with 0.5-5.0% ammonia water, and concentrating to prepare a xinaomycin mother solution, or adding a carrier and drying to obtain powder. The invention provides a novel low-cost method for preparing a novel biological pesticide xinaomycin or preparation.

Description

technical field [0001] The invention relates to the technical field of biopesticide manufacture, in particular to a method for separating and extracting neoaumycin, a fermentation metabolite of actinomycetes. Background technique [0002] Active compounds derived from microorganisms are the main source of high-efficiency and low-toxicity agricultural antibiotics. Antibiotics developed from microbial metabolites are an important part of biological pesticides, and are also the main research direction of pesticide development in the world. [0003] The research on agricultural antibiotics in my country started earlier. In the early stage, my country successfully developed kasugamycin, Qingfengmycin, Jinggangmycin, polyoxin, princess mycin, pleiotropycin, Nongkang 120 and so on. Over the past ten years, some new agricultural antibiotics such as Zhongshengmycin, Wuyimycin, Ningnanmycin, Huaguangmycin, and pyrimycin have been developed. At present, in addition to Jinggangmycin, s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07H19/06C07H1/08C12P19/38C12R1/57
Inventor 周金燕杨杰钟娟谭红
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S