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Biological transformation method by utilizing hydrophobic compound of cyclodextrin

A biotransformation and cyclodextrin technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of long biotransformation time, limited bioavailability, low conversion efficiency, etc.

Inactive Publication Date: 2008-04-30
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above research results have effectively improved the biotransformation efficiency of cortisone acetate, because the method involves the preparation of microbial embedding carriers and the embedding of microbial cells and other process links, the conversion process is relatively complicated, and the above-mentioned process The application has a certain restriction and influence
[0007] The transformation of hydrophobic compounds 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.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Compound C of cortisone acetate 1,2 dehydrogenation reaction

[0077] Microbial strain

[0078] The microbial strain adopts Arthrobacter sp.CICC 10193, which can realize the C 1,2 The conversion of cortisone acetate to prednisone acetate by dehydrogenation.

[0079] Incline medium (g / L): glucose 10, yeast extract 10, agar 20, pH7.0

[0080] Seed medium (g / L): glucose 10, corn steep liquor 10, peptone 5, dipotassium hydrogen phosphate 2.5, pH 7.0

[0081] Fermentation medium (g / L): glucose 10, corn steep liquor 15, peptone 5, dipotassium hydrogen phosphate 2.5, pH 7.0

[0082] Cultivation of slant seeds: take a ring of slant strains and transfer to fresh slant medium, and culture at 32°C for 24 hours.

[0083] Bacterial culture:

[0084] Take 5mL of sterile water to wash the slanted seeds to prepare a bacterial suspension. Transplant into 30mL seed medium according to 1% inoculum amount, culture at 32°C and shake at 160r / min for 24h.

[0085] The seed culture sol...

Embodiment 2

[0090] Embodiment 2 is basically the same as Example 1

[0091] Compound C of cortisone acetate 1,2 dehydrogenation reaction

[0092] The conversion substrate cortisone acetate is ground into a fine powder, and is directly thrown into the fermentation medium at a concentration of 4% of the weight of the fermentation broth, then 5% industrial alcohol by volume of the fermentation medium is added, and the substrate acetic acid can be added simultaneously. The sulfobutyl ether-β-cyclodextrin with a pine molar ratio of 0.3:1 was fermented at 34° C. and 160 r / min for 50 h.

[0093] Heat the culture solution to a slight boil, stop the transformation, and let it cool down to room temperature naturally for analysis.

[0094] External standard method detection. The conversion rate was determined to be 98.7%. The conversion rate of the blank control sample without adding was 92.6% in the simultaneous experiment under the same conditions.

Embodiment 3

[0095] Embodiment 3 is basically the same as Example 1

[0096] Compound C of cortisone acetate 1,2 dehydrogenation reaction

[0097] The conversion substrate cortisone acetate is ground into a fine powder, and is directly thrown into the fermentation medium at a concentration of 2.5% of the weight of the fermentation broth, and then 4% industrial alcohol by volume of the fermentation medium is added, and the substrate acetic acid can be added simultaneously. The hydroxypropyl-β-cyclodextrin with a pine molar ratio of 0.25:1 was fermented at 34° C. and 160 r / min for 50 h.

[0098] Heat the culture solution to a slight boil, stop the transformation, and let it cool down to room temperature naturally for analysis.

[0099] External standard method detection. The conversion rate was determined to be 98.9%. The conversion rate of the blank control sample without adding was 93.7% in the simultaneous experiment under the same conditions.

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PUM

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Abstract

The invention discloses a bioconversion method by using cyclodextrin and the hydrophobic compound of the derivative thereof, belongs to the bioconversion technical field, in particular relates to the bioconversion method by using the cyclodextrin and the hydrophobic compound of the derivative thereof, and provides the bioconversion method by using the cyclodextrin and the hydrophobic compound of the derivative thereof. The method of the invention is particularly suitable for the microbial conversion of hydrophobic compounds, can effectively improve the dissolubility of reaction substrate in aqueous solution, improve the utilization rate and the reaction rate of the substrate, release the inhibition of the substrate and the product to a certain extent, and improve the conversion rate of the product. The invention adopts a conversion system including the cyclodextrin, the special structure of the cyclodextrin ensures the cyclodextrin to have a certain solubilizing power, and strengthens the bioavailability of the substrate.

Description

technical field [0001] The invention belongs to the technical field of hydrophobic compound biotransformation. It specifically relates to a method for biotransformation of hydrophobic compounds using cyclodextrin and its derivatives and its application. Background technique [0002] Biotransformation refers to the process of using enzymes or organisms (cells, organelles) as catalysts to achieve chemical transformation, also known as biocatalysis, which is a biological system (including bacteria, fungi, plant tissues, animal tissue culture systems or enzyme preparations of biological systems) A chemical reaction in which a source substrate is structurally modified. Biocatalysts include enzymes and cells. The biological enzymes with industrial application value are mainly derived from microbial strains. For multi-enzyme systems and biocatalysis with coenzymes, whole cells of microorganisms are often used to participate in the reaction, also known as microbial transformation r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/02C12P33/06C12R1/06C12R1/32C12R1/645
Inventor 王敏杜连祥骆健美申雁冰肖克胜王建新
Owner TIANJIN UNIV OF SCI & TECH
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