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Method for preparing anthracene Ensamu bacterial

A technology of trace elements, Streptomyces nocardia, applied in microorganism-based methods, biochemical equipment and methods, antibacterial drugs, etc. problems such as low price and large output fluctuation, to achieve the effects of improved extraction yield and finished product quality, high proportion of effective components and low water content

Active Publication Date: 2007-05-02
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Patents related to the report of anthrasamectin include U.S. Patents US4265814, US42643913, WO0345312, WO0177360, WO0366005, WO0430731, WO0115119, and the Japanese Antibiotic Journal (1981), which describe the relevant physical and chemical properties, curative effect, traditional fermentation and extraction separation, etc. However, according to the culture medium reported in the literature, the high-yield genotype cannot be fully expressed due to the characteristics of the bacteria produced, the fermentation process, and the control conditions, resulting in low fermentation titers and yield fluctuations of anthrasamectin Disadvantages such as large, unstable, and high cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: preparation anthrasamcin

[0026] (1) Preparation and cultivation of slant spores

[0027] Ratio of slant spore medium (%): yeast extract 0.2, malt extract 0.3%, peptone 0.1, glycerol 1.0, agar powder 2, pH 7.0 before sterilization, test tube 30×200mm, capacity 20ml, heated at 121°C After sterilizing for 30 minutes, cool to 50-60°C and set aside the slope, then insert the spores and cultivate them at 29±1°C for 5-7 days to mature.

[0028] (2) Preparation and cultivation of seed solution

[0029] Seed medium ratio (%): starch 2, soybean peptone 0.6, corn steep liquor 0.5, CaCO 3 0.5, soybean cake powder 1.0, glucose 1, NaCl0.3, pH 6.8 before elimination, culture temperature 29±1°C, 220rpm, shaker culture for 36-48 hours, then pH 7.0-8.0, mycelium concentration is 30-50% (note: shake bottle 500ml specification filling volume 50ml, slant spore inoculation volume is 1×1cm 2 , The filter medium for seed culture is 8 layers of gauze.

[0030] (3) Preparati...

Embodiment 2

[0032] Embodiment 2: preparation anthrasamcin

[0033] (1) Preparation of seed liquid in seed tank

[0034] In the seed tank of 20L, drop into the seed medium of 9L (see embodiment 1 for the proportioning of the seed medium, and simultaneously add 0.3% soybean oil as an anti-foaming agent), sterilize with damp heat at 121°C for 30 minutes, after cooling , insert 100ml shake flask seed liquid, stir 150rpm, aeration ratio is 1: 1V / V / m, cultivate 36-48 hours, at this moment PH 7.0-8.0, mycelia concentration is 30-50%.

[0035] (2) Preparation and cultivation of fermenter culture medium

[0036] The proportion of the fermentation medium is the same as that of the aforementioned Example 1, but 0.05% PPG is also added as an antifoaming agent. The fermenter is 30L, the feeding volume is 18L, the pH before elimination is 6.7, and the pH after elimination is about 7.0. Sterilize for 30 minutes, after cooling, add about 1.8L of seed tank culture solution, then at 29±1°C, stir at 150-200...

Embodiment 3

[0037] Embodiment 3: preparation anthrasamcin

[0038] See Example 1 for the preparation of inclined plane spores and seed solution.

[0039] The formula composition of the fermentation medium (%): starch 5, glucose 1, lactose 1, soybean cake powder 2, peptone 1, corn steep liquor 0.5, CaCO 3 0.5, NaCl0.1, isobutanol 0.2, NiCl 2 0.001, MgSO 4 The volume of 0.002, 500ml shaker bottle is 40ml, and the sterilization condition is 121°C for 30 minutes, in which isobutanol is added in a sterile state after sterilization. Then 2ml of seed solution was added, and cultured on a shaking table at 29±1°C and 220rpm for 7-8 days. After the fermentation was completed, the titer of anthrasamectin was measured to be 660r / ml by high performance liquid phase.

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PUM

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Abstract

This invention provides a preparation method of high producing fermenting Anthracene Brassica Bacterin, and it uses S.nocardia fermenting under adequate fermentation medium containing mixed liquor with a certain proporion trace elements to produceAnthracene Brassica Bacterin. The above-mentioned fermentation medium contains 6-8% available carbon source and 3-6% available nitrogen source, whose precursor is 0.15-0.45% isobutanol. The biosynthetic level oAnthracene Brassica Bacterin is normal and stable because of the change in charge ratio and composition, as well as the adjustment of partial process controlling condition. The yield increases by 3 times comparing with old preparation and the finished product improves greatly.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a method for high-yield fermentation production of anthrasamectin. Background technique [0002] Ansamitocin (ansamitocin) is an ANSA antibiotic, and its main active ingredient is ansamitocin P3 produced by Streptomyces nocardia (S.Nocardia) during the liquid submerged fermentation process, which has many Strong cytotoxicity and antitumor activity, is a class of broad-spectrum antitumor antibiotics, and has strong antibacterial activity against Gram-positive bacteria and mycobacteria, and also has a certain effect on Gram-negative bacteria and viruses. Filamentous fungi and protozoa show a strong effect, and its mechanism of action is to inhibit the synthesis of DNA, RNA and protein in cells. It has been clinically shown to have significant curative effect on various lymphoid leukemias and malignant lymphomas. The molecular weight of its anthrasamterin P3 is 635.2, and its mole...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18A61P31/04A61P35/00C12R1/365
Inventor 蒋世春许玉丽白骅
Owner ZHEJIANG HISUN PHARMA CO LTD