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Strain for efficiently transforming saponin in turmeric to produce diosgenin, and application thereof

A diosgenin and high-efficiency technology, which is applied in the efficient transformation of turmeric saponins to produce diosgenin, and the layered Fusarium spore field, can solve the problems of inability to industrially produce, many by-products, and low microbial transformation efficiency.

Active Publication Date: 2012-10-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Diosgenin is usually extracted from turmeric (Dioscorea shield leaf) by acid hydrolysis, but this method has brought very serious environmental pollution, and the production of diosgenin by microbial transformation combined with enzymatic hydrolysis is generally considered to be the future The trend of diosgenin production has become one of the hot spots in the field of cleaner production
[0003] Many researchers have found that some microorganisms of the genus Trichoderma and Aspergillus can convert saponins in turmeric into diosgenin. Although the biotransformation method has mild conditions and is environmentally friendly, the conversion efficiency of these microorganisms is low and there are many by-products. really enter industrial production
Because the saponins in turmeric are complex, and most of them are fat-soluble compounds with low water solubility, the substrate feeding concentration and conversion efficiency are very low during the conversion process. These problems are difficult problems in the production of diosgenin by biotransformation.
Therefore, it is of great significance to screen a microorganism that efficiently transforms and produces diosgenin. However, there is no report on the research on the transformation and production of diosgenin by Fusarium spp. A big advantage of production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1 Enzymatic hydrolysis method to obtain turmeric saponin.

[0015] Weigh 100 g of dry turmeric powder, add it to 2000 mL of water, boil for 1 h, cool down, adjust the pH to 6.5, add 0.4 mL of medium-temperature amylase, and incubate at 65 °C for 1 h. After cooling, adjust the pH to 6.0, add 0.4 mL of thermostable amylase, and incubate at 80 °C for 1 h. Cool down, adjust the pH to 5.5, add 1 mL of liquid glucoamylase, and incubate at 60 °C for 8 h. Cool, centrifuge at 4000 rpm for 20 min, take the precipitate, dry at 60°C until constant weight, and weigh.

Embodiment 2

[0016] Embodiment 2 Utilizes layer out fusarium ( Fusarium proliferatum ) WX12 transformation to produce diosgenin.

[0017] (1) Preparation of Fusarium exfoliates ( Fusarium proliferatum ) Liquid culture of cells from the WX12 strain.

[0018] Pick Fusarium exfoliates ( Fusarium proliferatum ) One loop of WX12 strain, inoculated in a 250 mL Erlenmeyer flask containing 30 mL of seed medium, placed on a shaker at 200 r / min at 30°C and cultivated for 20-24 h to the logarithmic phase, that is Fusarium exophyllum ( Fusarium proliferatum ) Liquid culture of cells from the WX12 strain.

[0019] (2) The composition and ratio of the fermentation medium are: sucrose 20 g / L, ammonium sulfate 2 g / L, sodium nitrate 2 g / L, sodium chloride 5 g / L, dipotassium hydrogen phosphate 1 g / L, Magnesium sulfate heptahydrate 0.5 g / L, potassium chloride 0.5 g / L, iron sulfate heptahydrate 0.01 g / L, water 1000 mL, pH natural, sterilized under high-pressure steam at 121°C for 20 min.

[0020] (3) Sh...

Embodiment 3

[0022] Example 3 The diosgenin produced by the conversion was extracted with petroleum ether.

[0023] Centrifuge the transformation liquid at 8000-12000 r / min for 10 min, take the precipitate, add petroleum ether (boiling point 60 ℃ ~ 90 ℃) 20 mL ultrasonic extraction twice, 4 h each time, combine the extracts, and concentrate at 80 ℃ to about 15 mL. The obtained petroleum ether solution of diosgenin has a concentration of diosgenin of about 0.25 mg / mL and a purity of >40%.

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Abstract

The invention belongs to the biological technical field and particularly relates to a screening method of a high-yield diosgenin strain and an application of the strain in turmeric saponin transformation. With the adoption of the screening method, a Fusarium Proliferatum WX12 of high-yield diosgenin is screened; the strain is preserved in General Microbial Center of China Microbial Culture Collection Management Committee; the preservation number is CGMCCN0. 5901; the strain has characteristics of fast growth speed and high transformation efficiency, is applied in industrially producing the diosgenin, has the advantages of moderate reaction condition, high yield, low production cost and environment friendliness, and is convenient and quick in post treatment because a large number of acid or alkali is not needed.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a strain of Fusarium exfoliate ( Fusarium proliferatum ) WX12 and a method for efficiently transforming saponins in turmeric to produce diosgenin by using the bacteria. Background technique [0002] Diosgenin is an important basic raw material for the production of steroid hormone drugs. Steroid hormones have strong anti-infection, anti-allergic, anti-viral and anti-shock pharmacological effects, and are important drugs for treating rheumatism, cardiovascular, lymphatic leukemia, cellular encephalitis, skin diseases, anti-tumor and rescuing critically ill patients. It is the second largest class of drugs after antibiotics. At present, more than one thousand steroid hormone drugs are synthesized or semi-synthesized from diosgenin, and China is one of the most important producers of diosgenin. Diosgenin is usually extracted from turmeric (Dioscorea shield leaf) by acid hydrolysis, b...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P33/20C12R1/77
Inventor 史劲松张佳佳李恒李会张旦旦许正宏
Owner JIANGNAN UNIV
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