Method for separating and purifying water-soluble antibiotic Zwittermicin A in Bacillus thuringiensis
A technology for separation and purification of thuringiensis spores, applied in chemical instruments and methods, preparation of organic compounds, organic chemistry, etc., can solve the problems of expensive high-performance liquid chromatography instruments, complicated experiments, and many steps in purification methods, and achieve accurate results. Easy to operate and avoid the effect of oxidation
Inactive Publication Date: 2011-11-23
GUILIN UNIVERSITY OF TECHNOLOGY
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Problems solved by technology
The above-mentioned purification method has many steps, complicated tests, high-performance liquid chromatography equipment is relatively expensive, and professional technicians are needed at the same time, and there is no standard product of Zwittermicin A so far
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[0015] Centrifuge the fermentation broth of Bacillus thuringiensis subsp at 10,000r / min for 10 minutes to obtain the supernatant of the fermentation broth, and concentrate the aqueous phase of the supernatant to 50 times in vacuum at 50°C; according to the volume ratio, add 1 / 2 volume of absolute ethanol, then centrifuge at 10000r / min for 10min, discard the precipitated phase, keep the water phase; add an equal volume of petroleum ether to the water phase, shake well, let stand, separate the water phase and petroleum ether phase, discard the petroleum Ether phase, keep the water phase; the supernatant sample that has been preliminarily purified by absolute ethanol and petroleum ether is collected for later use; 2 mL of the carrier electrolyte liquid with a pH of 6.5, 1 mL of the supernatant sample, a drop of bromophenol blue, 7 mL of distilled water, Mix 0.1g of agarose, raise the temperature of the mixed glue to 80°C to fully dissolve, keep warm and pour the glue into a medic...
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The invention discloses a method for separating and purifying water-soluble antibiotic Zwittermicin A in Bacillus thuringiensis through an isoelectrofocusing electrophoresis method, which comprises the following steps: preliminarily purifying the supernate of a Bacillus thuringiensis fermentation liquid with anhydrous alcohol and petroleum ether, and sampling; preparing the supernate sample, an amphoteric electrolyte liquid, bromphenol blue, distilled water and agarose into a gel liquid, heating to 75-85 DEG C until the materials are sufficiently dissolved, and pouring the liquid into a medical rubber tube with a diameter of about 0.3cm while keeping the temperature; cooling the liquid for two hours, and introducing into an electrophoresis cell; and respectively adding electrolyte into negative and positive poles as required, and immersing both ends of the rubber tube into the electrode liquid to carry out isoelectrofocusing electrophoresis, thus separating and purifying the water-soluble antibiotic Zwittermicin A. As the experiment agents and samples are isolated from the air in the invention, the oxidation is avoided, the result is more accurate, the method is simple to operate, and the antibiotic Zwittermicin A can effectively separated out.
Description
technical field [0001] The invention belongs to the fields of phytochemistry, agricultural chemistry, biomedicine, plant protection, food chemistry, analytical chemistry, organic chemistry and biochemistry, and particularly relates to a method for separating and purifying water-soluble antibiotic Zwittermicin A in Bacillus thuringiensis. Background technique [0002] At present, the chemical insecticides widely used in the world not only kill pests, but also kill a large number of natural enemies of pests, resulting in the destruction of ecological balance and serious pollution of the environment, which poses a huge threat to human life. In addition, pests have gradually developed resistance to long-term use of chemical pesticides, forming a vicious cycle of chemical pesticide application. Biopesticides can partially overcome the above shortcomings. Therefore, actively developing and applying biopesticides to replace chemical pesticides to prevent and control pests on fruit...
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IPC IPC(8): C07C275/16C07C273/18
Inventor 郝再彬李洋刘红艳严丽张杰宋福平
Owner GUILIN UNIVERSITY OF TECHNOLOGY