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Method for promoting efficient conversion of Colletotrichum lini to DHEA (dehydroepiandrosterone) by adding ethylene glycol

A technology of Trichospora flaxensis and DHEA, which is applied in the biological field to achieve the effects of high substrate conversion rate, high product yield, and high substrate feeding concentration

Inactive Publication Date: 2017-04-26
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in the process of DHEA hydroxylation by Collectotrichum lini (Collectotrichum lini) ST-1, 16 kinds of common organic solvents were selected for analysis, and it was found that adding ethylene glycol can significantly improve the conversion rate of DHEA and the total yield of produ

Method used

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  • Method for promoting efficient conversion of Colletotrichum lini to DHEA (dehydroepiandrosterone) by adding ethylene glycol
  • Method for promoting efficient conversion of Colletotrichum lini to DHEA (dehydroepiandrosterone) by adding ethylene glycol
  • Method for promoting efficient conversion of Colletotrichum lini to DHEA (dehydroepiandrosterone) by adding ethylene glycol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Adding ethylene glycol, using Collectotrichum lini (Collectotrichum lini) ST-1 to hydroxylate DHEA

[0020] (1) Preparation of a cell liquid culture of the flaxenum (Collectotrichum lini) ST-1 strain

[0021] Pick a loop of Collectotrichum lini ST-1 strain on the solid PDA medium, inoculate it in a 250mL Erlenmeyer flask with 30mL seed medium, place it on a shaker at 200rr at 30°C Cultivate at a rotational speed of 20-24 hours to the logarithmic phase to obtain a cell liquid culture of Collectotrichum lini ST-1 strain.

[0022] (2) The composition and ratio of the fermentation medium are: glucose 20-50g / L; yeast powder 10-30g / L; peptone 5-20g / L; corn steep liquor 1-10g / L; pH 6.5-7.5, 121°C Sterilize under high pressure steam for 20min.

[0023] (3) Shake flask fermentation: inoculate the liquid culture of cells of the above-mentioned Collectotrichum lini ST-1 strain in a sterilized 30mL fermentation tank with an inoculum of 6-10% (w / w) The culture medium is...

Embodiment 2

[0025] After embodiment 2 adds ethylene glycol, to the influence of thorn sporodium flaxensis (Collectotrichum lini) ST-1 hydroxylation DHEA

[0026] (1) Substrate conversion rate after adding ethylene glycol

[0027] In the 3L liquid fermentation medium, while adding the substrate DHEA, 15mL of glycerin was added. At the end of the fermentation, the conversion rate of the substrate in the obtained fermentation broth was as high as 90.4%, and the conversion rate of the substrate in the control example without adding ethylene glycol It is 67.5%, which is 33.9% higher than the control example without adding ethylene glycol.

[0028] (2) Conversion rate of products 7α-OH-DHEA and 7,15α-diOH-DHEA after adding ethylene glycol

[0029] In the 3L liquid fermentation medium, while adding the substrate DHEA, 15mL of glycerin was added. At the end of the fermentation, the product yields of the products 7α-OH-DHEA and 7,15α-diOH-DHEA in the resulting fermentation broth increased to 86.8...

Embodiment 3

[0030] Embodiment 3 adds 16 kinds of organic solvents to the influence of substrate DHEA biotransformation reaction

[0031] The applicant selected a variety of common organic solvents, and found that there was no significant improvement in the substrate biotransformation in the scheme of the present invention. Only after adding 0.5% ethylene glycol, the applicant unexpectedly found that no matter the substrate conversion rate , or the yield of 7,15α-diOH-DHEA, the total yield of 7α-OH-DHEA and 7,15α-diOH-DHEA were significantly increased.

[0032] The Influence of Organic Solvents on the Conversion of DHEA

[0033]

[0034]

[0035]

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Abstract

The invention relates to a biotechnology field and relates to a method for improving hydroxylation of Colletotrichum lini ST-1 to DHEA (dehydroepiandrosterone) using ethylene glycol. The method is characterized in that 0.5% (v/v) of ethylene glycol is directly added while the substrate DHEA is added during the conversion, thereby significantly improving the conversion efficiency of the DHEA. Under the condition that the feeding amount of the substrate is 12g/L, the conversion of the substrate is up to 90.4%, and the total yield of the product 7alpha-OH-dehydroepiandrosterone and 7,15alpha-diOH-dehydroepiandrosterone is 86.8%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method of adding ethylene glycol to increase the hydroxylated dehydroepiandrosterone (DHEA) of Colletotrichum lini ST-1 to 7α-hydroxy-dehydroepiandrosterone (7α-OH-DHEA) and 7,15α-hydroxy-dehydroepiandrosterone (7,15α-diOH-DHEA) methods. Background technique [0002] Some major 3-hydroxy steroids such as dehydroepiandrosterone (DHEA), cholesterol, and pregnenolone will be transformed into corresponding 7α-hydroxy steroid derivatives in natural organisms. In recent years, the important biological activities of DHEA, 7α-hydroxy-DHEA and 7,15α-hydroxy-DHEA have attracted more attention. At present, studies have found that 7α-OH-DHEA can prevent hypoxic cell death of neurons in vitro, and has obvious antioxidant and anti-tumor effects. It can be used as a hydroxysteroid with important pharmacological effects and medicinal value. body compound; and the dihydroxylation prod...

Claims

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Application Information

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IPC IPC(8): C12P33/06C12R1/645
Inventor 李会许正宏史劲松李聪倪瑜张晓梅
Owner JIANGNAN UNIV
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