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High-concentration cholesterol culture medium as well as preparation method and application thereof

A cholesterol, high-concentration technology, applied in the field of cholesterol degradation research, can solve the problems of complicated medium configuration process, inability to complete the configuration, 2-3 days, etc., to achieve uniform and stable distribution of cholesterol, not easy to precipitate, and long storage time. Effect

Pending Publication Date: 2021-06-01
JIANGXI SCI & TECH NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, according to "Cholesterol-lowering Probiotic Lactic Acid Bacteria Screening and Research on Its Functional Mechanism" and "Phospholipid and cholesterol uptake by mycoplasma cells and membranes", the cholesterol medium used for traditional culture of microorganisms or cells is lecithin-cholesterol micellar medium, which requires Dissolve lecithin and cholesterol in chloroform, then blow dry with nitrogen, and then perform processes such as ultrasonic crushing and centrifugation, and the prepared medium needs to be stored in a 4°C refrigerator for a maximum of 2-3 days
The preparation process of this medium is complicated, it cannot be prepared in a short time, the storage time is short, and the materials used are expensive, so it is only suitable for the preparation of milligram-level low-concentration cholesterol medium

Method used

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  • High-concentration cholesterol culture medium as well as preparation method and application thereof

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Effect test

Embodiment 1

[0030] A cholesterol sterile CD liquid medium, the content of effective components of the medium includes: sodium nitrate 0.2g / 100mL, potassium dihydrogen phosphate 0.1g / 100mL, magnesium sulfate 0.05g / 100mL, potassium chloride 0.05g / 100mL , sodium chloride 0.05g / 100mL, ferrous sulfate 0.002g / 100mL, cholesterol 1.5g / 100mL and water.

[0031] Its preparation method concrete steps are:

[0032] S1. Accurately weigh sodium nitrate, potassium dihydrogen phosphate, magnesium sulfate, potassium chloride, sodium chloride, and ferrous sulfate with an electronic balance, add deionized water to dissolve, pack in a triangular flask, and seal;

[0033] S2. Sterilize the sealed triangular flask containing the medium in an autoclave at a temperature of 121°C for 20 minutes, and store it at 4°C to obtain a sterile CD liquid medium. stand-by;

[0034] S3. Spread the weighed cholesterol on a class 100 ultra-clean bench, close the glass window of the ultra-clean bench, and carry out ultraviole...

Embodiment 2

[0039] A cholesterol sterile CD solid medium, the effective component content of the medium includes: sodium nitrate 0.2g / 100mL, potassium dihydrogen phosphate 0.1g / 100mL, magnesium sulfate 0.05g / 100mL, potassium chloride 0.05g / 100mL , sodium chloride 0.05g / 100mL, ferrous sulfate 0.002g / 100mL, agar 1.5g / 100mL, cholesterol 1.5g / 100mL and water.

[0040] Its preparation method concrete steps are:

[0041] S1. Accurately weigh sodium nitrate, potassium dihydrogen phosphate, magnesium sulfate, potassium chloride, sodium chloride, ferrous sulfate, agar, and deionized water to dissolve with an electronic balance, pack in a triangular flask, and seal;

[0042] S2. Sterilize the sealed triangular flask containing the medium in an autoclave at a temperature of 121°C for 20 minutes, and store it at 4°C to obtain a sterile CD solid medium. stand-by;

[0043] S3. Spread the weighed cholesterol on a class 100 ultra-clean bench, close the glass window of the ultra-clean bench, and carry o...

Embodiment 3

[0048] Add 100 μ L of Aspergillus oryzae spore suspension in the high-concentration cholesterol CD liquid medium of embodiment 1 (cholesterol concentration is 1.5g / 100ml, loading is 100mL) and grows, will add the cholesterol CD medium of Aspergillus oryzae spore suspension , placed in a shaker at 30°C for 96 hours of fermentation, the appearance photos of its growth at different times are as follows figure 1 Shown, where a is 0h; b is 48h; c is 72h; d is 96h. Depend on figure 1 It can be seen that as time increases, the medium changes from turbid to clear after 48 hours, because cholesterol is insoluble in water, so it shows that cholesterol in high-concentration cholesterol medium decreases with time, and Aspergillus oryzae gradually utilizes and metabolizes cholesterol. Illustrate that the high-concentration cholesterol CD liquid medium provided by the present invention can be used for research in the field of cholesterol, and in this medium, Aspergillus oryzae has higher m...

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Abstract

The invention provides a high-concentration cholesterol culture medium as well as a preparation method and application thereof, and belongs to the technical field of cholesterol degradation research. The invention provides a high-concentration cholesterol culture medium, the content of cholesterol in the culture medium can reach 3g / 100mL, the culture medium not only contains nutrient substances required by the growth of aspergillus oryzae, but also has relatively good dispersity and stability, and the culture medium is suitable for cholesterol degradation research tests of microorganisms, especially aspergillus oryzae. The invention also provides a preparation method of the high-concentration cholesterol culture medium, the use of an organic solvent is not involved in the preparation method, the preparation steps are simple and rapid, and the cholesterol in the prepared culture medium is uniformly distributed and cannot be oxidized and degraded when being used under normal test conditions. Tests prove that the culture medium can be used for related experimental research on cholesterol degradation of microorganisms, especially aspergillus oryzae.

Description

technical field [0001] The invention belongs to the technical field of cholesterol degradation research, and in particular relates to a high-concentration cholesterol medium and its preparation method and application. Background technique [0002] Cholesterol is an essential substance for the human body, accounting for 0.2% of the total body weight. When cholesterol is excessive in the body, it will lead to hypercholesterolemia, arteriosclerosis, coronary heart disease, stroke and other diseases, which have seriously threatened human health. At present, scholars at home and abroad have more and more researches on microbial degradation of cholesterol. In the study of microbial degradation of cholesterol, the configuration of high-concentration cholesterol medium is an important link. However, according to "Cholesterol-lowering Probiotic Lactic Acid Bacteria Screening and Research on Its Functional Mechanism" and "Phospholipid and cholesterol uptake by mycoplasma cells and me...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/60C12Q1/02C12R1/69
CPCC12N1/14C12Q1/60C12Q1/02G01N2333/38
Inventor 曾斌邱尚昆
Owner JIANGXI SCI & TECH NORMAL UNIV
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