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Preparation method for Bacillamide compound and Bacillamide precursor

A compound and precursor technology, applied in the field of bacterial culture to prepare compounds, can solve problems such as co-cultivation system instability and poor repeatability, and achieve the effects of low industrial production cost, high repeatability, and cost reduction

Active Publication Date: 2014-04-16
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a way to increase the expression of microbial secondary metabolites, co-cultivation still has many problems in its application. For example, there is no stimulating effect between strains, or there is an effect of stimulating the expression of metabolites, but the co-cultivation system is unstable and poor in repeatability, etc.

Method used

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  • Preparation method for Bacillamide compound and Bacillamide precursor
  • Preparation method for Bacillamide compound and Bacillamide precursor
  • Preparation method for Bacillamide compound and Bacillamide precursor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 1) Inoculate actinomycetes (Streptomyces nodosus1A0101684, Beijing Beina Chuanglian Biotechnology Research Institute) into 500mL Erlenmeyer flasks, each containing 200mL of fermentation medium (based on 1L of fermentation medium, yeast extract 5g, glycerin 4g, soluble starch 1g, and seawater as the balance, adjust the pH of the fermentation medium to 7.5 with an acid-base buffer solution, and divide it into four groups, ABCD, wherein group A and group B were cultured statically at 28°C for 6 Day, Group C and Group D were cultured statically at 28°C for 10 days;

[0051] 2) Inoculate the Bacillus mycoides CGMCC1.197, which was cultivated on a shaker at 150 rpm for 24 hours, from Beijing Beina Chuanglian Biotechnology Research Institute, into the fermentation culture of the grown actinomycetes in step 1) Among them, the inoculation volume of group A and C was 400 μL, and the inoculation volume of group B and D was 1000 μL, and the symbiotic static culture was carried out ...

Embodiment 2

[0057] 1) Inoculate actinomycetes (Streptomyces nodosus1A0101684, Beijing Beina Chuanglian Biotechnology Research Institute) into 500mL Erlenmeyer flasks, each containing 250mL fermentation medium (based on 1L of fermentation medium, yeast extract 5g, glycerin 5g, and seawater as the balance, adjust the pH of the fermentation medium to 7.5) with an acid-base buffer solution, and culture it statically at 28°C for 7 days;

[0058] 2) Inoculate the Bacillus mycoides CGMCC1.197, which was cultured on a shaker at 140 rpm for 24 hours, from Beijing Beinan Chuanglian Biotechnology Research Institute, into the culture medium in which actinomycetes have grown in step 1) In , the inoculation volume was 1000 μL, and the symbiosis was statically cultured at 28°C for 15 days to obtain the co-cultivation fermentation medium;

[0059] 3) The co-cultivation fermentation medium obtained in each group was extracted with ethyl acetate and concentrated to dryness to obtain ethyl acetate fractions...

Embodiment 3

[0073] The ethyl acetate part obtained in Example 2 was separated by reverse phase chromatography to obtain a bacillamide compound and a bacillamide precursor. The filler used in the reversed-phase chromatography separation method is octadecyl bonded silica gel, and the mobile phase used is methanol-water. The specific process of the reversed-phase chromatography separation method is as follows: firstly use methanol aqueous solution formed by volume ratio 30:70 methanol and water to remove large polar impurities, and then use methanol aqueous solution formed by volume ratio 40:60 to obtain Bacillamide precursor, then eluted with methanol aqueous solution formed by methanol and water at a volume ratio of 50:50 to obtain bacillamide compounds B and C (eluted in time order), and eluted with methanol aqueous solution formed by 60:40 methanol and water to obtain bacillamide compound a.

[0074] Based on the fermentation medium, the yield of bacillamide precursor was 20 mg / L, the y...

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Abstract

The invention discloses a method for large-scale production of a Bacillamide compound and a Bacillamide precursor. The method comprises the steps as follows: actinomycetes are inoculated onto a fermentation culture medium and cultured for 5-10 days; bacillus mycoides are also inoculated onto the fermentation culture medium and subjected to symbiotic culture for 10-20 days to obtain a co-cultured fermentation culture medium, and the Bacillamide compound and the Bacillamide precursor are obtained after separation. The method provided by the invention is low in the requirement on a culture medium, simple to operate, and low in industrial production cost, the fermentation product is simple, high in the content of effective compositions and low in the content of purities, and not only lowers the cost, but also can be in large-scale production to obtain the Bacillamide compound and the Bacillamide precursor, and very high application value and high industrial production feasibility are achieved.

Description

technical field [0001] The invention relates to the field of bacterial culture and preparation of compounds, in particular to a preparation method for mass production of bacillamide compounds and bacillamide precursors. Background technique [0002] Red tides not only seriously damage the marine ecological environment, marine fisheries and marine resources, but also various toxins brought by red tides, such as diarrhea shellfish toxins, amnesia shellfish toxins, paralytic shellfish toxins, etc., seriously endanger human health. For the control of algal blooms in red tide and aquaculture water, many methods have been proposed at home and abroad, which can be divided into physical methods, biological methods and chemical methods. The physical method generally uses the flocculation of modified clay to settle the algae, but this method treats the symptoms but not the root cause, and the algae will multiply in large numbers. The biological method uses compound microbial preparat...

Claims

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Application Information

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IPC IPC(8): C12P17/14C12P17/10C12R1/04
Inventor 马忠俊余立雁
Owner ZHEJIANG UNIV
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