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Method for preparing Chinese yam saponin by microorganism transformation process

A technology for microbial transformation and diosgenin, which is applied in the field of microbial transformation of diosgenin in plants into diosgenin, which can solve the problems of increased production costs, expensive equipment and operating costs, and large investment in wastewater treatment, so as to reduce raw materials Waste, reduce production cost, improve the effect of yield

Inactive Publication Date: 2010-09-15
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In recent years, improved chemical preparation methods of diosgenin include ethanol crystallization (CN1188425C), super (near) critical water hydrolysis (CN1896093), vaporized acid acid hydrolysis (CN1709904), and the use of new equipment (CN1182152C) for production, but these methods all can't get rid of the acid hydrolysis process, although there are also the disclosure of the patents of the non-wastewater production method (CN1850852) and the pollution-free production method (CN1850853) of diosgenin, the treatment cost of waste water and pollutants is huge, and will undoubtedly increase production cost
The supercritical fluid extraction method disclosed in Chinese patent CN1240791 does not use an acid hydrolysis process, but the equipment and operating costs are expensive, which restricts its application in production
Chinese patent CN 85108564B introduces a method for co-producing citric acid and diosgenin by fermenting Aspergillus niger. In this method, Dioscorea shield leaf is first fermented by Aspergillus niger, and then acid hydrolysis and solvent extraction are carried out. The yield of diosgenin can reach 4 %, but the patent failed to get rid of the acid hydrolysis process, the investment in wastewater treatment was huge, and the comparison effect between the fermentation method of Aspergillus niger and the acid hydrolysis method was not disclosed, and it was impossible to correctly evaluate the microorganism in the case of large differences in the content of Dioscorea scutellaria in different production areas conversion efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Medium: A. Seed medium: 30ml of bran extract (10g of bran plus 100ml of tap water boiled for 20min, filtered, and the filtrate was added to 100ml with water). B. Fermentation medium: 0.9g (60 mesh) of Dioscorea scutellaria powder, 0.1g of urea, KH 2 PO 4 0.03g, bran extract 30ml, sterilized at 121°C for 20min.

[0023] The slant surface of Aspergillus oryzae CICC 2436 was inoculated on the seed culture based on 25°C, 24 hours of cultivation, the fermentation culture was carried out in a 250ml shake flask, the actual liquid volume was 30ml, the inoculation amount was 10%, the speed was controlled at 150rpm, and the temperature was controlled at 30°C . After the fermentation was carried out for 48 hours, the culture temperature was changed to 50° C., and the culture was continued for 2 hours.

[0024] After the cultivation, add 95% ethanol twice the volume of the fermentation broth, microwave-assisted extraction, add 1% medical activated carbon for decolorization, evap...

Embodiment 2

[0027]Medium: A. Seed medium: 30ml of bran extract. B. Fermentation medium: transformation substrate pangolin powder 0.9 g (60 mesh), urea 0.08 g, KH 2 PO 4 0.03g, bran extract 30ml, sterilized at 121°C for 20min.

[0028] The slant surface of Aspergillus oryzae CICC 2436 was inoculated on the seed culture based on 25°C, 24 hours of cultivation, the fermentation culture was carried out in a 250ml shake flask, the actual liquid volume was 30ml, the inoculation amount was 8%, the speed was controlled at 220rpm, and the temperature was controlled at 25°C After 60 hours of fermentation, the culture temperature was changed to 60° C., and the culture was continued for 3 hours.

[0029] After the cultivation, add 80% methanol twice the volume of the fermentation broth, microwave-assisted extraction, add 1% medical activated carbon for decolorization, evaporate ethanol, extract with an appropriate amount of No. 120 gasoline, concentrate, and use an appropriate amount of 90 The pure...

Embodiment 3

[0032] Culture medium: A. Seed medium: bran extract. B. Fermentation medium: 2500ml of bran extract, 75g of the transformation substrate Dioscorea scutellaria, 7.5g of urea, KH 2 PO 4 3g, sterilized at 121°C for 20min.

[0033] The slant surface of Aspergillus oryzae CICC 2436 was inoculated on the seed culture based on the seed medium, the liquid volume was 30mL, the shaking table speed was 220rpm, the culture temperature was 30°C, and the culture time was 24 hours; the fermentation culture was carried out in a 5L fermenter, and the actual loading The liquid volume is 2.5 L, the inoculum volume is 10%, the rotational speed is controlled at 250 rpm, the air volume is 1 vvm, and the temperature is controlled at 30°C. After the fermentation was carried out for 60 hours, the culture temperature was changed to 50° C., and the culture was continued for 2 hours.

[0034] After the fermentation is finished, add 95% ethanol 1 times the volume of the fermentation broth, and microwav...

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Abstract

The invention discloses a transmitting method from dioscin into diosgenin saponin in the biological conversing and purifying technical domain of natural product, which comprises the following steps: adding carbon source and nitrogen source to form seed culture medium and fermenting culture medium; seeding aspergillus oryzae; aerating gas to stir; fermenting; extracting ferment liquid assisted by microwave; improving receiving rate by 1.8-2.2 times corresponding to acid hydrolytic method.

Description

technical field [0001] The invention belongs to the technical field of biotransformation and extraction of natural products, and particularly relates to a method for converting diosgenin in plants into diosgenin with microorganisms. Background technique [0002] Diosgenin (Formula I), which is a derivative of isospirostane, is a diosgenin in the rhizomes of Dioscoreaceae plants, and its glycosides are usually glucose, rhamnose, galactose, arabinose, etc. Dioscin has the effect of estrogen, which can lower cholesterol, relieve cough, eliminate phlegm, desensitize, restore diseased tissue, and stimulate heparin cells and bile secretion. It is an ideal precursor for the synthesis of various steroid hormones and steroidal contraceptives. [0003] Dioscorea saponins widely exist in plants in nature, especially in Dioscoreaceae. Among them, Dioscorea zingiberensis wright, D. nipponica Makino, D. parviflora Ting Sp. Nov, D. Panthaica Prain et. (D. nipponica Makno subsp. Deltoide...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/20C12R1/69
Inventor 修志龙董悦生刘琳齐珊珊王辉张代佳
Owner DALIAN UNIV OF TECH
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