Method for preparing sterol derivative through Burkholderia conversion and application

A technology of Burkholderia and Krkholderia, applied in the field of Burkholderia transformation and preparation of sterol derivatives

Inactive Publication Date: 2020-07-31
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The Burkholderia screened in our laboratory can not only produce cholesterol esterase and cholesterol oxidase, but also degrade cholesterol, sitosterol, st...

Method used

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  • Method for preparing sterol derivative through Burkholderia conversion and application
  • Method for preparing sterol derivative through Burkholderia conversion and application
  • Method for preparing sterol derivative through Burkholderia conversion and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035]Example 1: α-cyclodextrin system B.cepacia converts cholesterol to prepare cholest-4-ene-3,6-dione

[0036] (1) Slant culture: Take Burkholderia ZWS15 stored in a glycerol tube at -40°C, inoculate 30 μL into LB solid medium, place it in a constant temperature incubator at 37°C, and cultivate for 24 hours until a single colony grows;

[0037] (2) Seed culture: Pick a single colony from the strain cultivated in step (1) under sterile conditions and inoculate it in 50 mL LB liquid medium, and culture it in a shaker at 37°C with a rotation speed of 200 rpm for 9-13 hours to OD 600 5.0 to 6.0.

[0038] (3) B. cepacia conversion of cholesterol: the seed solution cultivated in step (2) was inserted into the transformation medium at an inoculum size of 5% by volume, and cultured at 30° C. and 200 rpm for 48 hours. The group without cyclodextrin or its derivatives was used as the control.

[0039] (4) TLC separation and extraction of the conversion product of cholesterol: the ...

Embodiment 2

[0045] Example 2: β-cyclodextrin system B.cepacia converts cholesterol to prepare cholest-4-ene-3,6-dione

[0046] See embodiment 1 for the specific embodiment.

[0047] The amount of fixed cholesterol in the transformation medium was 2g / L, that is, 5.18mM; the molar ratio of β-cyclodextrin to cholesterol was 1:2, 1:1 and 2:1. The two were pretreated with 60W ultrasonic for 10-15min and then added to the transformation medium.

[0048] Analysis and detection: When the molar ratio of β-cyclodextrin and cholesterol is 1:2, 1:1 and 2:1, the molar conversion rate of cholest-4-ene-3,6-dione is 0.38% respectively , 0.36% and 0.42%. The conversion rate of the control group under the same conditions was 0.2%.

Embodiment 3

[0049] Example 3: γ-cyclodextrin system B.cepacia converts cholesterol to prepare cholest-4-ene-3,6-dione

[0050] See embodiment 1 for the specific embodiment.

[0051] The amount of fixed cholesterol in the transformation medium was 5.18mM; the molar ratio of γ-cyclodextrin to cholesterol was 1:2, 1:1 and 2:1. The two were pretreated with 60W ultrasonic for 10-15min and then added to the transformation medium.

[0052] Analysis and detection: when the molar ratio of γ-cyclodextrin to cholesterol is 1:2, 1:1 and 2:1, the molar conversion rate of cholest-4-ene-3,6-dione is 0.51% respectively , 0.35% and 0.26%. The conversion rate of the control group under the same conditions was 0.2%.

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Abstract

The invention discloses a method for preparing a sterol derivative through Burkholderia conversion and application, and belongs to the technical field of sterol microbial conversion. Burkholderia is added into a reaction system containing cyclodextrin or a derivative thereof, so that the product conversion yield can be increased to 12-15% from 0.1-0.3% of a control group; and in addition, an organic solvent does not need to be added in the preparation process of the method, so that the method is more environment-friendly, economical and safe.

Description

technical field [0001] The invention relates to a method and application for preparing sterol derivatives by transformation of Burkholderia bacteria, and belongs to the technical field of sterol transformation by microorganisms. Background technique [0002] Cholesterol is an important sterol compound that can be used as a raw material for steroidal drugs. Some microorganisms can excise the saturated side chain of cholesterol and oxidize the specific part of the steroid nucleus, so as to obtain a variety of derivatives with medicinal value. Liu WH et al. used Mycobacterium sp. to convert cholesterol into testosterone (TS); Nagasawa et al. used Arthrobacter simplex to degrade cholesterol and successfully produced androsta-1,4-diene-3,17-dione (ADD); Guo Qian et al. used Rhodococcus sp. converts cholesterol to cholest-4-en-3-one. In addition to the above derivatives, cholesterol can also be converted by microorganisms into cholest-4-ene-3,6-dione. [0003] Cholester-4-ene-3...

Claims

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Application Information

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IPC IPC(8): C12P33/02C12R1/01
CPCC12P33/02
Inventor 张玲王武杨海麟武迪
Owner JIANGNAN UNIV
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