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Biological sterol transforming process with cyclodextrin and its application

A biotransformation and microbial transformation technology, applied in the field of sterol biotransformation, can solve the problems of low conversion efficiency, limited bioavailability, long biotransformation time, etc., and achieve the effect of increasing dissolution

Inactive Publication Date: 2007-08-22
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The transformation of hydrophobic compounds such as sterols by microorganisms is often severely restricted by some factors: the solubility of the substrate in water is very small, which limits its bioavailability, the product / substrate has an inhibitory effect on microorganisms, the biotransformation time is long, and the conversion efficiency is low. shortcoming

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Activation culture of microbial slant: Take a ring of slant bacteria and transfer to fresh slant, culture at 29°C for 3 days.

[0049] Activation culture of microbial cell liquid seeds: Wash the seeds cultivated on the slope with 0.5% Tween80 sterile aqueous solution to make the bacterial suspension A 600 The value was controlled at 1.0±0.2, and the inoculum was inserted into the transformation medium according to the inoculum size of 2%, and activated and cultivated at 27°C and 200r / min for 36h.

[0050] Phytosterol conversion: liquid seeds are inserted into the transformation medium (pre-added 1% phytosterol) by 5% inoculum size, hydroxypropyl-β-cyclodextrin is added at 30 hours, and the culture solution is heated when it is cultivated for 72 hours When it boiled slightly, the transformation was stopped, and it was naturally lowered to room temperature for analysis.

[0051] Microbial strains using Mycob actorium sp.NRRL B-3683

[0052] Slant medium (g / L): 0.5 potas...

Embodiment 2

[0056] Embodiment 2 basic method is the same as example 1

[0057] Activation culture of microbial slant: Take a ring of slant strains and transfer to fresh slant, culture at 29°C for 3 days.

[0058] Activation culture of microbial cell liquid seeds: Wash the seeds cultivated on the slope with 0.5% Tween80 sterile aqueous solution to make the bacterial suspension A 600 The value was controlled at 1.0±0.2, and the inoculum was inserted into the transformation medium according to 2% of the inoculum amount, and activated and cultured at 30°C and 200r / min for 30h.

[0059] Phytosterol conversion: liquid seeds are inserted into the transformation medium (pre-added 0.5% phytosterol) according to the inoculum size of 5%, add sulfobutyl ether-β-cyclodextrin at 24h, and heat the culture solution when cultivating to 72h When it boiled slightly, the transformation was stopped, and it was naturally lowered to room temperature for analysis. The molar ratio of phytosterol to sulfobutyl e...

Embodiment 3

[0062] Embodiment 3 basic method is the same as example 1

[0063] The microorganisms were activated and cultured at 28°C and 200r / min for 36h, and 10% of the inoculum was inserted into the transformation medium (with 1% phytosterol added in advance), and hydroxypropyl-β-cyclodextrin was added at 36h, the molar ratio It is hydroxypropyl-β-cyclodextrin: phytosterol = 2:1. When the culture reaches 84 hours, heat the culture solution to a slight boil, stop the conversion, and naturally cool down to room temperature for analysis. The molar ratio of phytosterol to hydroxypropyl-β-cyclodextrin is 1:2,

[0064] Phytosterols consist of sitosterol, stigmasterol and campesterol in a 2:1:1 ratio. The microbial strain used was Mycobacterium sp.NRRL B-3683. The conversion rate was 81.9%, compared to 27.7% for the control experiment.

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PUM

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Abstract

The present invention belongs to the field of biological sterol transforming technology, and is especially biological sterol transforming process with cyclodextrin and its derivative and its preparation. In a transforming system, cyclodextrin or its derivative in certain amount is added, or substrate and cyclodextrin or its derivative in certain ratio are made to form inclusion compound, so as to facilitate transformation. The process of the present invention is especially suitable for microbial transformation of hydrophobic compound, and can raise the solubility of reaction substrate in water solution, raise the substrate utilizing rate and reaction rate, release substrate and product inhibition and raise product converting rate. When the substrate concentration is 3-20g / L, the present invention has conversion period shorted to 3-4 days, conversion rate of 50-95 % and high industrial application value.

Description

technical field [0001] The invention belongs to the technical field of sterol biotransformation. It specifically relates to a sterol biotransformation method utilizing cyclodextrin and its derivatives and its application. Background technique [0002] Sterol is a substance with cyclopentane polyhydrophenanthrene as the skeleton (also known as steroid nucleus), which exists in free and bound forms in nature. The appearance is white to off-white crystalline powder. Sterols can be divided into three categories: animal sterols, plant sterols and microbial sterols. Animal sterols are mainly cholesterol, and phytosterols are mainly sitosterol, stigmasterol and campesterol, etc., which exist in plant seeds. [0003] More than 80% of the raw materials for the synthesis of steroid drugs in the world are to obtain androst-4-ene-3,17-dione (AD) and androst-1,4-ene-3,17-dione (ADD) by sterol side chain degradation ), the AD and ADD obtained after microbial transformation of sterols ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P33/16C12R1/06C12R1/32
Inventor 王敏杜连祥骆健美申雁冰肖克胜
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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