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Substrate treatment method for phytosterol side chain degradation reaction

A technology for phytosterol and degradation reaction, which is applied in the field of substrate treatment of phytosterol side chain degradation reaction, can solve the problems of poor contact between the substrate and microbial cells, easy aggregation on the surface of the water phase, prolonged transformation time, etc. Achieving the effect of sufficient raw material sources, low price and improved conversion rate

Active Publication Date: 2016-08-31
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The selective degradation of phytosterol side chains by microorganisms is carried out under the action of steroidal invertase produced by microbial cells. The main problem encountered in the process of degrading phytosterol side chains to generate AD is that phytosterols have strong hydrophobicity. The solubility in water is extremely low, and it is easy to gather on the surface of the water phase, which makes the substrate and microbial cells not in good contact, resulting in a series of problems such as low conversion rate and prolonged conversion time.

Method used

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  • Substrate treatment method for phytosterol side chain degradation reaction
  • Substrate treatment method for phytosterol side chain degradation reaction
  • Substrate treatment method for phytosterol side chain degradation reaction

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0023] 1 Preparation of co-modified attapulgite-phytosterols:

[0024] (1) Acid co-modified attapulgite-phytosterol: add phytosterol and attapulgite to 100mL of 4mol / L acid solution in a mass ratio of 1:0.1~5, wherein the total mass of phytosterol and attapulgite does not exceed 5g, the acid solution is one of sulfuric acid, hydrochloric acid, nitric acid or phosphoric acid, ultrasonically oscillated for 30min, centrifuged and washed until neutral, dried at 60°C for 8h, and ground into fine powder.

[0025] (2) Silane coupling agent co-modified attapulgite-phytosterol: add phytosterol and attapulgite to 100mL of acetic acid aqueous solution with pH 3.5 according to the mass ratio of 1:0.1~5, the total mass of phytosterol and attapulgite is not More than 5g, then add 0.1mL of silane coupling agent, the silane coupling agent is 3-aminopropyltriethoxysilane KH550, ultrasonic vibration for 30min, centrifugal washing until no silane coupling agent, dry at 60℃ for 8h, grind into po...

Embodiment 1

[0036] Embodiment 1: silane coupling agent co-modifies attapulgite-phytosterol

[0037] 1 Modification of the substrate:

[0038] Add 1g phytosterol PS and 3g attapulgite AT to 100mL acetic acid solution (pH3.5), then add 0.1mL 3-aminopropyltriethoxysilane KH550, ultrasonically shake for 30min, centrifuge at 5000r / min for 10min, deionize Wash with water until there is no KH550, dry at 60°C for 8 hours, and grind into a fine powder, which is silane coupling agent co-modified attapulgite-phytosterol KH550-AT-PS.

[0039] 2 Degradation reactions of phytosterol side chains

[0040] The silane coupling agent co-modified attapulgite-phytosterol KH550-AT-PS was added to the mycobacterium fermentation medium for reaction.

[0041] The components of the fermentation medium are: glucose 10g / L, citric acid 2g / L, ferric ammonium citrate 0.05g / L, magnesium sulfate 0.5g / L, dipotassium hydrogen phosphate 0.5g / L, diammonium hydrogen phosphate 3.5g / L L, pH 7.2.

[0042] The inoculum amount...

Embodiment 2

[0053] Example 2: Co-modification of attapulgite-phytosterols with acid

[0054] 1 Modification of the substrate:

[0055] Add 1g of phytosterol PS and 3g of attapulgite AT into 100mL of 4mol / L sulfuric acid solution, ultrasonically oscillate for 30min, centrifuge at 5000r / min for 10min, wash with deionized water until neutral, dry at 60°C for 8h, and grind into fine powder. Sulfuric acid co-modified attapulgite-phytosterol H 2 SO 4 -AT-PS.

[0056] 2 Degradation reactions of phytosterol side chains

[0057] co-modified attapulgite-phytosterol H with sulfuric acid 2 SO 4 -AT-PS was added to the mycobacterial fermentation medium for reaction.

[0058] The components of the fermentation medium are: glucose 10g / L, citric acid 2g / L, ferric ammonium citrate 0.05g / L, magnesium sulfate 0.5g / L, dipotassium hydrogen phosphate 0.5g / L, diammonium hydrogen phosphate 3.5g / L L, pH 7.2.

[0059] The inoculum amount of mycobacteria was 7% (v / v).

[0060] The fermentation system of ad...

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Abstract

The invention belongs to the field of microbial pharmaceutics and provides a substrate treatment method for phytosterol side chain degradation reaction. The method is applied to phytosterol side chain degradation reaction for AD (androstenedione) production. Phytosterol and attapulgite are modified prior to feeding of phytosterol, and according to a substrate feeding quantity and an attapulgite addition amount, a mass ratio of the phytosterol to the attapulgite is controlled to be 1:0.1-5, and concentration of fed co-modified phytosterol and attapulgite is 1-40g / L. The substrate treatment method has advantages that dispersity of a substrate in an aqueous solution is effectively improved, the utilization rate and the reaction rate of the substrate are increased, and substrate and product inhibition is relieved to some extent to increase the conversion rate of products; in addition, the method is simple in process, economical, practical, low in environment pollution and significant to biotransfomation based steroid medicine production.

Description

technical field [0001] The invention belongs to the field of microbial pharmacy, and in particular relates to a substrate treatment method for phytosterol side chain degradation reaction. Background technique [0002] Steroid hormone drugs are an important category in the field of medicine in my country, and Androstenedion (AD) is an irreplaceable intermediate of steroid hormone drugs. my country is a big oil producing country, and there are extremely rich phytosterol resources in the by-products of the oil industry. Microorganisms selectively degrade phytosterol (phytosterol, PS) side chains to generate AD, which can replace complex multi-step chemical synthesis methods, and alleviate the current shortage of resources caused by diosgenin as raw materials. The development of the pharmaceutical industry is of great significance. [0003] The selective degradation of phytosterol side chains by microorganisms is carried out under the action of steroidal invertase produced by ...

Claims

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Application Information

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IPC IPC(8): C12P33/16C12P33/02C12R1/32
CPCC12P33/005C12P33/02
Inventor 申雁冰王敏刘晶晶汤睿骆健美郑宇
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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