Method for producing coenzyme Q10 by enzyme engineering method

A technology using enzymes and coenzymes, applied in quinone separation/purification, fermentation, etc., can solve the problems of coenzyme Q10, such as no relevant reports and inconvenience, and achieve the effects of improving synthesis efficiency, improving product quality, and reducing production costs

Inactive Publication Date: 2009-12-09
SHANGHAI FANGYI BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, there are no relevant reports at home and abroad on the production of coenzyme Q10 by using enzyme engineering methods
[0009] This shows that the above-mentioned existing method for producing coenzyme Q10 obviously still has inconvenience and defects in the method, and needs to be further improved urgently.
In order to solve the problems existing in the method of producing coenzyme Q10, relevant manufacturers have tried their best to find a solution, but no suitable design has been developed for a long time, and there is no suitable way to solve the above problems in general methods

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example

[0035] The polymerase was prepared by Rhodopseudomonas photosynthetic bacteria.

[0036] 1) Species selection: Select Rhodopseudomonas photosynthetic bacteria under a microscope. The shape of the bacteria is short rod-shaped or egg-shaped, with a width of 0.5-0.9 microns and a length of 1.2-2.0 microns. It has no spores, no capsule, and monopolar flagella. It is a healthy and mobile strain.

[0037] 2) Preparation of fermentation broth: 5.5g sodium dihydrogen phosphate, 1.5gDL malic acid, 2g sodium acetate, 2g sodium hydroxide, 1g ammonium chloride, 0.25g magnesium chloride, 0.05g calcium chloride, 3.5g glucose dissolved in 1 liter of distilled water , Stir evenly, and the measured pH should be between 6.5 and 7.0.

[0038] 3) Take 10 250mL Erlenmeyer flasks, inject 100mL culture solution into each after sterilization, and seal them after inoculation. Place them in a light incubator at 32°C for 72 hours and take them out.

[0039] 4) Collect the culture medium and centrifug...

Embodiment 1

[0041] Using coenzyme Q1 and solanesol as raw materials, coenzyme Q10 is synthesized under the action of polymerase.

[0042]1) Coenzyme Q1 phosphorylation. Under the condition of 30°C, dissolve 250g of coenzyme Q1 in 1.5L of phosphate buffer (pH=7.4), add 12.5g of phospholipid according to 5% of the weight of coenzyme Q1, and stir for 30 minutes;

[0043] 2) Bromination of solanesol. Under the condition of 5°C, dissolve 631g of solanesol in 3L of ether, add 63mg of phosphorus trichloride according to 0.01% of the weight of solanesol, and stir for 150 minutes;

[0044] 3) According to the method of the above-mentioned polymerase preparation example, prepare the polymerase for use;

[0045] 4) Add the prepared coenzyme Q1 and solanesol into the reaction vessel under the condition of 40°C-60°C, add 86 mg of the prepared polymerase, and stir for 10-30 minutes;

[0046] 5) Add 4.5 L of n-hexane, stir and extract at 30° C. for 60 minutes. After resting, take the n-hexane layer....

Embodiment 2

[0052] Using coenzyme Q0 and polydecaprene as raw materials, coenzyme Q10 is synthesized under the action of polymerase.

[0053] 1) Coenzyme Q0 phosphorylation. Under the condition of 30°C, dissolve 172g of coenzyme Q0 in 1.2L of phosphate buffer, add 8.6g of phosphate ester according to 5% of the weight of coenzyme Q0, and stir for 30 minutes;

[0054] 2) Phosphorylation of polydecaisoprene. Under the condition of 25°C, 709 g of polydecaisoprene was dissolved in 3.3 L of phosphate buffer, and adenosine triphosphate was added according to 0.01% of the weight of polydecaisoprene, and stirred for 120 minutes;

[0055] 3) According to the method of the above-mentioned polymerase preparation example, prepare the polymerase for use;

[0056] 4) Under the condition of 40°C-60°C, add the prepared coenzyme Q0 and polydecaisoprene into the reaction vessel, add 86 mg of polymerase, and stir for 10-30 minutes;

[0057] 5) Add 4.5 L of n-hexane, stir and extract at 30° C. for 60 minut...

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PUM

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Abstract

The present invention relates to a method for producing coenzyme Q10 using an enzyme engineering method, using coenzyme Q0 and decapolyisoprene as raw materials, or using coenzyme Q1 and solanesol as raw materials, wherein Phosphorylation of polyisoprene, bromination of solanesol; under the action of polymerase, under the condition of 40 ℃ ~ 60 ℃, the synthesis reaction of coenzyme Q10 is carried out in the reactor, followed by extraction, concentration and crystallization , and dried to obtain the finished product. The polymerase is obtained through the following steps: selecting Rhodopseudomonas photosynthetic bacteria containing coenzyme Q10 and culturing; breaking the bacterial cells and precipitating with 3% saline; separating the precipitate to obtain the polymerase. The invention has specificity, improves the reaction efficiency and improves the product quality. Coenzyme Q10 produced by the enzyme engineering method has no isoprene with cis structure in its side chain, and the biological activity of the product is the same as that of the product produced by the microbial fermentation method, and the production cost is greatly reduced.

Description

technical field [0001] The invention relates to a method for producing coenzyme Q10, in particular to a method for producing coenzyme Q10 by using an enzyme engineering method. Background technique [0002] Coenzyme Q10, also known as ubiquinone 10, is an over-the-counter drug included in the Pharmacopoeia in my country. Coenzyme Q10 is a fat-soluble quinone, which is orange-yellow crystal at room temperature, and its melting point is 48°C. It is odorless and tasteless. It is named after the degree of polymerization of pentadiene group is 10, and it is a quinone ring compound. The physiological function of coenzyme Q10 mainly comes from the oxidation-reduction properties of the quinone group and the physical properties of the isoprenoid side chain. It has two main functions in the human body. One is to have an obvious anti-lipid peroxidation effect; Nutrients play an important role in the conversion of nutrients into energy in the mitochondria. Its quinone ring plays the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/66C07C46/10
Inventor 熊军胡秋滨马开龙
Owner SHANGHAI FANGYI BIO ENG
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