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Method for extracting coenzyme Q10 from microorganism

A technology of microorganisms and coenzymes, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of low extraction rate, unsuitable for batch sample application, and time-consuming, etc., and achieve high extraction rate, reagents and The effect of rapid operation steps and low cost

Active Publication Date: 2012-07-04
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The laboratory separation technology reported in Japanese related literature is simply using organic solvents for long-term extraction, which is too time-consuming and the extraction rate is too low, so it is not suitable for batch sample applications

Method used

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  • Method for extracting coenzyme Q10 from microorganism
  • Method for extracting coenzyme Q10 from microorganism
  • Method for extracting coenzyme Q10 from microorganism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031]Embodiment 1, extract coenzyme Q from Rhodobacter sphaeroides 10

[0032] Coenzyme Q is extracted from Rhodobacter sphaeroides by ultrasonic crushing method of the present invention 10 , the specific method includes the following steps:

[0033] 1. Fermentation and cultivation of Rhodobacter sphaeroides

[0034] Rhodobacter sphaeroides ATCC 17023 (purchased from the American Type Microorganism Collection) was added to the medium shown in Table 1. The medium is mixed with the components in Table 1, the final volume is adjusted to 1 liter, the pH is adjusted to 7.0 (pH6-8 is acceptable), and then autoclaved at 121° C. for 20 minutes. For solid media, mix the above ingredients with 15 g of agar, then autoclave, cool the media to about 60°C, and pour into sterilized Petri dish plates. Stir under aerobic conditions, ferment and cultivate for 28 hours at a temperature of 25-35° C. (18-72 hours are all available). After the cultivation, 4 g of wet bacterial cells were obta...

Embodiment 2

[0053] Embodiment 2: Alkaline alcohol saponification method extracts coenzyme Q 10 And HPLC content detection (control)

[0054] 1. Alkaline alcohol saponification to extract coenzyme Q 10

[0055] Take 4g of fermented product of Rhodobacter sphaeroides ATCC 17023, add 3g of pyrogallic acid, stir evenly, then add 5g of sodium hydroxide and 26mL of methanol aqueous solution, stir evenly, saponify at 90°C for 1 hour, after cooling rapidly, add 50mL of n-hexane, vigorously After oscillating, let stand to separate layers, extract twice, take the extract, wash with deionized water until neutral, then remove water with anhydrous sodium sulfate, 35°C rotary evaporation under reduced pressure, redissolve in absolute ethanol to obtain coenzyme Q 10 The crude extract was stored at -20°C until testing.

[0056] 2. HPLC content detection

[0057] Carry out HPLC analysis with the method identical with step 2, the result is as follows Figure 4 As shown, coenzyme Q appears at the rete...

Embodiment 3

[0058] Embodiment 3: Coenzyme Q is extracted by alkaline lysis 10 And HPLC content detection

[0059] Coenzyme Q is extracted from Rhodobacter sphaeroides ATCC 17023 fermentation product by alkaline lysis method of the present invention 10 , the specific method includes the following steps:

[0060] Preparation of alkaline lysate:

[0061] Solution I: Accurately weigh 0.9908g glucose, 0.3028g Tris Cl, 0.3722g EDTA, dissolve them in distilled water, adjust the volume to 100mL, adjust the pH to 8.0, sterilize at 121°C for 20min, and put them into 10mL sterile EP tubes , stored at 4°C for later use.

[0062] Solution II: Accurately weigh 0.4000g NaOH and 0.5000g SDS, dissolve them in distilled water, dilute to 50mL, and set aside.

[0063] Solution III: Accurately weigh 14.7225g of potassium acetate, dissolve in distilled water, add 5.75mL of glacial acetic acid, make up to 50mL with distilled water, divide into 10mL sterile EP tubes, and store at 4°C for later use.

[0064]...

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Abstract

The invention discloses a method for extracting coenzyme Q10 from microorganisms, wherein the ultrasonic disintegration is to mix microorganisms with the ability of producing the coenzyme Q10 and an organic solvent in which the coenzyme Q10 is well dissolved according to the mass volume ratio of 1-5 to 10-20, and to obtain the coenzyme Q10 after ultrasonication at a temperature of 0 DEG C, wherein the ultrasonic frequency is between 0.2 and 0.8 and the power is between 300 and 500 watts; and the alkaline splitting method is to extract the coenzyme Q10 from fermentation broth of Rhodobacter sphaeroides. The microorganisms are preferred to be microorganisms of the Rhodobacter sphaeroides. The method for extracting the coenzyme Q10 has the advantages of high efficiency, low pollution, low requirements on apparatuses and equipment, low cost and so on, and is suitable for promotion and application.

Description

technical field [0001] The present invention relates to coenzyme Q 10 Extraction method, particularly involving a method of extracting coenzyme Q from microorganisms 10 Methods. Background technique [0002] coenzyme Q 10 (Coenzyme Q 10 , Ubiquinone) is widely present in animal and plant tissues and microbial cells. It is an important hydrogen transporter in the respiratory chain and an essential coenzyme for generating ATP. coenzyme Q 10 As a natural antioxidant and cell activator, it has the functions of directly improving human immunity, enhancing antioxidant capacity, and delaying aging. coenzyme Q 10 Clinically, it is mainly used in the treatment of cardiovascular diseases, and it can also be used in the comprehensive treatment of cerebral edema caused by viral hepatitis, subacute liver necrosis and fulminant hepatitis. coenzyme Q 10 Congenital deficiency (clinically divided into four categories: myopathies with myoglobinuria and central nervous system diseases;...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/26C07C50/28C12R1/01
Inventor 于群李伟静贺敏宋丽雅乔志新任鹏
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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