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Method for producing glycyrrhetinic acid through microbial fermentation transformation

A technology for microbial fermentation and glycyrrhetic acid, applied in the field of microorganisms, can solve the problems of not yet forming a large-scale level, and the microbial transformation of glycyrrhetic acid is not much, so as to improve the utilization rate of raw materials, reduce waste liquid discharge and environmental pollution, and improve products. Effects of yield and purity

Active Publication Date: 2013-11-20
中科医药行业生产力促进中心有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there are not many research reports on the microbial transformation of glycyrrhetinic acid abroad. Domestic research institutions such as Sichuan University, Beijing University of Traditional Chinese Medicine, and Hubei University of Technology conduct related research, but they are still in the stage of experimental research and have not yet formed a large-scale level.

Method used

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  • Method for producing glycyrrhetinic acid through microbial fermentation transformation
  • Method for producing glycyrrhetinic acid through microbial fermentation transformation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Inoculate Aspergillus flavus (Aspergillus flavus) CGMCC No5144 into the Erlenmeyer flask containing the seed medium, the pH is natural, the inoculum size is 1%, the rotation speed is 200rpm, and the ventilation volume is 0.2m 3 / h, and the incubation time was 24 hours.

[0041] Transfer the above cultured strains into a 100L fermenter (with 60L of sterilized fermentation medium inside) according to the inoculum amount of 30%, adjust the fermentation temperature to 10°C, and the ventilation volume to 0.4m 3 / h, stirring speed 150rmp, fermentation 3d.

[0042] Investigate the production of glycyrrhetinic acid. Detection of glycyrrhetinic acid by vanillin sulfate method:

[0043] (1) Reference substance solution preparation

[0044] Accurately weigh 20 mg of glycyrrhetinic acid standard product dried at 100°C to constant weight, put it in a 20 ml measuring bottle, add absolute ethanol to the mark, and obtain a glycyrrhetinic acid standard solution with a content of 1 mg...

Embodiment 2

[0050] Inoculate Aspergillus flavus (Aspergillus flavus) CGMCC No5144 into the Erlenmeyer flask containing the seed medium, the pH is natural, the inoculum size is 1%, the rotation speed is 200rpm, and the ventilation volume is 0.2m 3 / h, and the incubation time was 24 hours.

[0051] Transfer the above cultured strains into a 100L fermenter (with 60L of sterilized fermentation medium inside) according to the inoculum amount of 30%, adjust the fermentation temperature to 20°C, and the ventilation volume to 0.4m 3 / h, stirring speed 150rmp, fermentation 3d, strain fermentation concentration reached 3.1g / L.

[0052] Glycyrrhetinic acid detection method is the same as above.

Embodiment 3

[0054] Inoculate Aspergillus flavus (Aspergillus flavus) CGMCC No5144 into the Erlenmeyer flask containing the seed medium, the pH is natural, the inoculum size is 1%, the rotation speed is 200rpm, and the ventilation volume is 0.2m 3 / h, and the incubation time was 24 hours.

[0055] Transfer the above cultured strains into a 100L fermenter (with 60L sterilized fermentation medium inside) according to the inoculum amount of 30%, adjust the fermentation temperature to 30°C, and the ventilation volume to 0.4m 3 / h, stirring speed 150rmp, fermentation 3d, bacterial strain fermentation concentration reaches 4.2g / L.

[0056] Glycyrrhetinic acid detection method is the same as above.

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Abstract

The invention provides a method for producing glycyrrhetinic acid through microbial fermentation transformation. The method comprises the following steps of: inoculating aspergillus flavus CGMCC No 5144 into a solid potato dextrose agar (PDA) slant culture medium to cultivate; inoculating 1 percent of the cultivated aspergillus flavus CGMCC No 5144 to a culture medium containing fresh seeds from the inclined surface to cultivate; inoculating 30 percent of the strains obtained from the seed culture medium to a fermentation tank containing a fresh fermentation culture medium and performing fermentation culture to obtain fermentation liquid containing the glycyrrhetinic acid; separating a glycyrrhetinic acid coarse product from the fermentation liquid by a membrane separation combined process; and preparing high-purity glycyrrhetinic acid by a purification process. In the glycyrrhetinic acid finally obtained by the method, the fermentation quantity reaches 4.38 g / L, the product purity is more than or equal to 98 percent, the conversion rate is more than or equal to 90 percent, and the technological scale level is reached.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a technology for transforming and producing glycyrrhetinic acid with microorganisms. Background technique [0002] Licorice is a traditional Chinese medicinal material in my country, known as "the old man of the country", sweet in taste and flat in nature. Guixin, lung, spleen, stomach. Invigorating the spleen and replenishing qi, clearing away heat and detoxification, eliminating phlegm and relieving cough, relieving spasm and relieving pain, harmonizing various medicines. For weakness of the spleen and stomach, lassitude and weakness, palpitation and shortness of breath, cough with copious phlegm, epigastric abdomen, extremity convulsion acute pain, carbuncle sore tumefacting virus. It can alleviate the toxicity and potency of drugs. Licorice is widely used in medicine, tobacco, food and other fields. Glycyrrhizic acid and glycyrrhetinic acid are the main active ingr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/42C12R1/67
Inventor 王福清徐莉张桐
Owner 中科医药行业生产力促进中心有限公司
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