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Method for cultivating halophilic microorganisms to produce ectoine and hydroxy ectoine

A technology for hydroxytetrahydropyrimidine and halophilic microorganisms is applied in the field of culturing halophilic microorganisms to produce tetrahydropyrimidine and hydroxytetrahydropyrimidine, and can solve the problems of inhibiting growth, decreasing the synthesis amount of tetrahydropyrimidine, affecting the yield of hydroxytetrahydropyrimidine, etc. , to achieve the effect of improving the synthesis rate

Inactive Publication Date: 2011-12-21
朱道辰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a derivative of ectoine, the synthesis of hydroxyectoine is induced by heat stimulation, and only a small amount is synthesized at the optimal growth temperature of halophilic bacteria. Usually, a larger amount of synthesis can only be achieved by increasing the growth temperature. For example, Halomonas ventosae DL7 is in Shaker culture in the nutrient medium test tube, the culture temperature was increased from 30°C to 37°C or 42°C, but increasing the growth temperature seriously inhibited its growth, and led to a serious decline in the synthesis of ectoine, which also affected the hydroxy tetrahydropyrimidine synthesis. Hydropyrimidine production

Method used

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  • Method for cultivating halophilic microorganisms to produce ectoine and hydroxy ectoine
  • Method for cultivating halophilic microorganisms to produce ectoine and hydroxy ectoine
  • Method for cultivating halophilic microorganisms to produce ectoine and hydroxy ectoine

Examples

Experimental program
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Effect test

Embodiment 1

[0029] 1. Take an inoculation loop and connect the Halomonas ventosae DL7 strain to a test tube containing 6ml of liquid nutrient medium on a clean bench, and culture it in a shaker at 240rpm at 30°C for 20h for activation.

[0030] 2. Into a 500ml Erlenmeyer flask containing 50ml of seed culture medium, insert 2.5ml of the above-mentioned activated bacterial solution, culture on a shaker at 240rpm at 30°C for 24h, and culture 3 flasks in parallel. Seed medium (1L): glucose 10g, sodium chloride 58.4g, ammonium chloride 2g, magnesium sulfate 2g, dipotassium hydrogen phosphate 0.6g, yeast extract 1g, peptone 1g.

[0031] 3. Insert 150ml of seed bacterial liquid in a 5L fermenter with 3L fermentation medium, 800 rpm, pH control at 7.0-7.2, and ventilation at 0.2-0.4v / v.min (per minute per volume of culture Based on the volume ratio of sterile air), start to feed after 10 hours of fermentation, keep the glucose concentration at about 20g / l, cultivate at 30°C for 18 hours, add NaCl...

Embodiment 2

[0036] 1. Take an inoculation loop and connect the Halomonas ventosae DL7 strain to a test tube containing 6ml of liquid nutrient medium on a clean bench, culture it in a shaker at 240rpm at 30°C for 18h for activation.

[0037] 2. Into a 500ml Erlenmeyer flask containing 50ml of seed culture medium, insert 2.5ml of the above-mentioned activated bacterial solution, and culture on a shaker at 240rpm at 30°C for 18h, and culture 3 flasks in parallel. Seed medium (1L): glucose 10g, sodium chloride 58.4g, ammonium chloride 2g, magnesium sulfate 2g, dipotassium hydrogen phosphate 0.6g, yeast extract 1g, peptone 1g.

[0038] 3. Insert 150ml of seed bacterial liquid in a 5L fermenter with 3L fermentation medium, 800 rpm, pH control at 7.0-7.2, and ventilation at 0.2-0.4v / v.min (per minute per volume of culture Based on the volume ratio of sterile air), start to feed after 10 hours of fermentation, keep the glucose concentration at about 20g / l, cultivate at 30°C for 20 hours, add NaCl...

Embodiment 3

[0043] 1. Take an inoculation loop and connect the Halomonas ventosae DL7 strain to a test tube containing 6ml of liquid nutrient medium on a clean bench, and culture it in a shaker at 240rpm at 30°C for 24h for activation.

[0044] 2. Into a 500ml Erlenmeyer flask containing 50ml of seed medium, insert 2.5ml of the above-mentioned activated bacterial solution, and culture on a shaker at 240rpm at 30°C for 20h, and culture 3 flasks in parallel. Seed medium (1L): glucose 10g, sodium chloride 58.4g, ammonium chloride 2g, magnesium sulfate 2g, dipotassium hydrogen phosphate 0.6g, yeast extract 1g, peptone 1g.

[0045] 3. Insert 150ml of seed bacterial liquid in a 5L fermenter with 3L fermentation medium, 800 rpm, pH control at 7.0-7.2, and ventilation at 0.2-0.4v / v.min (per minute per volume of culture Based on the volume ratio of sterile air), start to feed after 10 hours of fermentation, keep the glucose concentration at about 20g / l, cultivate at 30°C for 24 hours, add NaCl to ...

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Abstract

The object of the present invention is to provide a method for producing ectoine and hydroxy ecto by cultivating halophilic microorganisms with reasonable cost to improve the synthesis efficiency of ectoine and hydroxy ecto. The method for cultivating halophilic microorganisms of the present invention to produce hydroxyectoine comprises the following steps: (1) strain activation; (2) seed cultivation; (3) fermentation stage; (4) reducing the salt concentration of the culture medium by feeding Rapidly increase to 3-3.5M, and then continue to cultivate for 3-8h, so that the synthesis of ectoine is greatly increased; (5) raise the culture temperature from the optimal growth temperature to a higher temperature, continue to cultivate for 3-10h, and obtain a higher High yield of ectoine synthesis. In the invention, the fermentation inoculation strain is cultivated by means of osmotic pressure shock and heat shock in the fermentation process, and the synthesis rate of ectoine and hydroxy ectoine is greatly improved at a reasonable cost.

Description

technical field [0001] The invention relates to a production method of ectoine and hydroxy ectoine, in particular to a method for cultivating halophilic microorganisms to produce ectoine and hydroxy ectoine. Background technique [0002] Ectoine is a compatible solute synthesized by halophilic microorganisms in order to resist the high salt concentration in the outside world, and some halophilic microorganisms can synthesize ectoine derivatives - hydroxy ectoine. Compared with other known compatible solutes, ectoine and hydroxyectoine have better protection for cells, nucleic acids, enzymes, proteins, etc. under extreme conditions such as radiation, drying, freezing, high temperature, and high salt effect. [0003] Due to its special structure, ectoine or hydroxyectoine cannot be synthesized up to now, and can only be obtained through intracellular synthesis of microorganisms. According to public literature reports, the synthesis of ectoine is mainly produced by bacterial ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/12C12R1/01
Inventor 朱道辰吴建陈华友王成栋陈焱
Owner 朱道辰
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