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A strain of Burkholderia cepacia and its application

A Burkholderia and onion technology, applied in the field of microorganisms, can solve problems such as the separation and extraction of unfavorable sterol enone derivatives, the impact on the secretion and activity of cholesterol oxidase, and the toxicity of microorganisms

Active Publication Date: 2021-03-30
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above method is to add a small amount of sterol solubilizer (organic solvent) in the fermentation medium, which will cause poisoning to microorganisms, affect the secretion and activity of cholesterol oxidase, and is not conducive to the subsequent large-scale generation and subsequent separation and extraction of sterol enone derivatives.

Method used

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  • A strain of Burkholderia cepacia and its application
  • A strain of Burkholderia cepacia and its application
  • A strain of Burkholderia cepacia and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Screening of Burkholderia cepacia ZWS15 and identification of 16SrDNA

[0046] (1) Sample collection and processing

[0047] The oil-contaminated soil was collected from Karamay Oilfield in Xinjiang (soil sampling depth is 5-20cm), placed in the enrichment medium on a shaker at 180rpm for enrichment culture for 48h, and the temperature was 30°C.

[0048] Enrichment medium (g / L): glucose 20, yeast extract 10, KH 2 PO 4 0.05, KNO 3 1. MgSO 4 0.5, FeSO 4 0.01, NaCl 0.5, Cholesterol 1, Triton X-100 0.5% (v / v).

[0049] (2) Primary screening

[0050] After the enrichment culture of the soil sample, the enriched bacterial liquid was taken and cultured on the primary screening medium plate at 50°C for 3 days.

[0051] Primary screening medium (g / L): K 2 HPO 4 2, KCl 0.5, NaNO 3 2, TritonX-100 3.4mL, MgSO 4 100mM, and add 0.1% cholesterol as carbon source.

[0052] (3) Re-screening

[0053] Pick the colonies grown on the primary screening medium pla...

Embodiment 2

[0059] Example 2: Physiological and biochemical properties of Burkholderia cepacia ZWS15

[0060] Burkholderia cepacia ZWS15 were respectively inoculated in the culture medium with cholesterol, sitosterol, stigmasterol, ergosterol, cholesterol, pregnenolone and other sterol substances as the only carbon source, and cultured at 50°C for 48h, Burkholderia cepacia was found All cepacia ZWS15 could grow colonies, which indicated that Burkholderia cepacia ZWS15 could utilize these carbon sources.

[0061] Medium formula (g / L): K 2 HPO 4 2, KCl 0.5, NaNO 3 2, TritonX-100 3.4mL, MgSO 4 100mM, and add 0.1% mass volume ratio of sterol substances as carbon source.

[0062] The physiological and biochemical characteristics of Burkholderia cepacia ZWS15 are shown in Table 2.

[0063] Physiological and biochemical characteristics of the bacterial strains in Table 2

[0064]

[0065] Note: ND: Not sure; +: Indicates that microorganisms can degrade the substance; -: Indicates that...

Embodiment 3

[0066] Example 3: Preparation of cholesterol oxidase using Burkholderia cepacia ZWS15

[0067] (1) Incline culture: inoculate Burkholderia cepacia ZWS15 on LB solid medium, and culture statically for 20-28 hours at 37°C;

[0068] LB solid medium: Add 5 g of yeast powder, 10 g of peptone, 10 g of NaCl, and 20 g of agar powder per 1 L of distilled water.

[0069] (2) Seed cultivation: Pick a single colony of the strain cultivated in step (1) under aseptic conditions and inoculate it in 50 mL of LB liquid medium, and culture it in a shaker at 37° C. with a rotation speed of 200 rpm for 9-13 hours.

[0070] LB liquid medium: add 5 g of yeast powder, 10 g of peptone, and 10 g of NaCl for every 1 L of distilled water.

[0071] (3) Fermentation culture: take the activated seed solution in step (2), inoculate it into the fermentation medium at an inoculum size of 3%-10%, and cultivate it in a shaker at 30° C. with a rotation speed of 200 rpm for 20-28 hours.

[0072] Fermentation me...

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Abstract

The invention discloses a strain of Burkholderia cepacia and its application, belonging to the field of microorganisms. The Burkholderia cepacia ZWS15 was deposited in the China Center for Type Culture Collection on November 6, 2017, with the preservation number CCTCC NO: M2017661, and the preservation address is Wuhan University, Wuhan, China. The bacterium can use cholesterol as the sole carbon source, can grow at 50°C, tolerates 10% organic solvents, and the enzyme activity of cholesterol oxidase in the crude enzyme liquid obtained by its fermentation is as high as 200U / L. Using Burkholderia cepacia ZWS15 fermented crude enzyme solution to catalyze cholesterol in a two-phase system in the presence of organic solvents, the amount of cholest-4-en-3-one collected was about 5-11% of the added amount of substrate %, the amount of cholest-4-ene-3,6-dione is about 10-15% of the amount of substrate added.

Description

technical field [0001] The invention relates to a strain of Burkholderia cepacia and its application, belonging to the field of microorganisms. Background technique [0002] Sterol derivatives have various physiological functions and can be used in industries such as medicine, food and feed. The 5-en-3-one derivatives, 4-en-3-one derivatives, and 4-en-3,6-dione derivatives of sterols all have good functions of improving lipid metabolism and can be used as weight-loss drugs. It has significant curative effect in treating liver disease, atherosclerosis and anti-obesity. [0003] Enone derivatives of sterols are mainly obtained by chemical synthesis or natural product extraction. For example, cholest-4-ene-3,6-dione is made from cholesterol, which is chemically synthesized by Jones reagent oxidation or PCC / dichloromethane method, or derived from marine natural product extracts; stigmaster-4-ene -3,6-dione can be obtained from plant extracts; cholest-4-en-3-one can be obtaine...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/04C12P33/02C12R1/01
CPCC12N1/20C12N9/0006C12P33/02C12Y101/03006C12N1/205C12R2001/01
Inventor 张玲王武武迪杨海麟辛瑜孙柳青
Owner JIANGNAN UNIV
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