JPS59173088A - Preparation of coenzyme q - Google Patents

Abstract

PURPOSE:To prepare coenzyme Q in high purity and recovery, by repeating the extraction and adsorption of a material containing coenzyme Q. CONSTITUTION:A material containing coenzyme Q, e.g. animal or vegetable tissue, microbial cells, etc. is made to contact with a hydrophilic solvent such as propanol, butanol, acetone, etc. in the presence of water to extract the coenzyme Q, and the extract is treated with a hydrophobic adsorbent such as silica gel, porous synthetic resin, etc. to adsorb the coenzyme Q from the extract liquid. The extraction treatment and the adsorption treatment are repeated preventing the contact of the extraction raw material with the adsorbent, and the adsorption residue obtained by the adsorption treatment, is if necessary, reused as the extractant.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing coenzyme Q, and more particularly to a method for obtaining coenzyme Q by repeating two processes of extraction and adsorption from a coenzyme Q-containing material.

Coenzyme Q in coenzyme Q-containing materials can be extracted using a hydrophilic solvent, and the extraction rate is better in the presence of water with a water content of several wt% than in anhydrous extraction. It is known.

On the other hand, as the water content during extraction becomes higher than the optimum value, the solubility of coenzyme Q in the extractant decreases rapidly, and the extraction rate also decreases significantly. Therefore, in extraction under high water content,
Increasing the extraction rate requires a large amount of extractant. By the way, concentrates or suspensions of coenzyme Q-containing substances usually contain a large amount of water, but when using substances containing a large amount of water as raw materials, extraction and mixing are necessary. The liquid has a high water content and requires a large amount of extractant.

To efficiently extract coenzyme Q from substances with high water content using a small amount of solvent, coenzyme Q is extracted by adding a hydrophilic solvent and an adsorbent together to a coenzyme Q-containing material and stirring A method is already known in which the contained substances are extracted and at the same time adsorbed onto an adsorbent (Japanese Patent Application Laid-Open No. 55-39
701 issue). However, in this method, before eluting coenzyme Q from the adsorbent, the extraction residue must usually be separated and removed from the adsorbent, but since both the adsorbent and the extraction residue are solids, it is necessary to remove the extraction residue substantially. Complete separation and removal is technically difficult and takes a short amount of time. Therefore, as a result of various studies regarding the method for producing coenzyme Q, the present inventors found that, instead of allowing the extraction raw material and the adsorbent to coexist, the extract was first obtained, and then the extract and the hydrophobic adsorbent were mixed together. The present invention has been completed in which after coenzyme Q is adsorbed on an adsorbent by contacting the adsorbent, the residual adsorption liquid is reused as an extractant if desired.

That is, the present invention provides a method for producing coenzyme Q from a coenzyme Q-containing material, including an extraction treatment in which a hydrophilic solvent and a coenzyme Q-containing material are brought into contact in the presence of water, and The adsorption treatment in which coenzyme Q in the coenzyme Q extract is adsorbed onto a hydrophobic adsorbent is repeated, and the adsorption residual liquid obtained from the adsorption treatment is optionally used as an extractant (7). This is a method for producing coenzyme Q.

In the present invention, coenzyme Q-containing substances include animal and plant tissues, and microbial cells such as bacteria, yeast, and filamentous fungi. Prior to extraction, movement.

Plant tissues are usually pretreated using a homogenizer, ultrasound, acid/alkali, or the like.

Further, among microorganisms, cells such as yeast and filamentous fungi are usually subjected to the above-mentioned pretreatment, but bacterial cells do not need to be subjected to pretreatment, but may be subjected to pretreatment.

After culturing, microbial cells are usually used in the form of wet microbial cells in the form of a paste or water suspension, and dried microbial cells obtained by drying these. However, it is not prohibited to use the culture solution as it is.

Hydrophilic solvents include methanol, ethanol, 7'q
/<Alfur such as alcohol and butanol, and solvents such as acetone.

Among these solvents, propatool, butanol, and acetone are usually used alone, and methanol and ethanol are also used singly, but usually propatool, min (wt)/water molecule solvent (wt)
50A'wt% or less, preferably 3 to 50W,
Particularly preferably, the water content is 5 to 407t%. Note that when water is low, it may be necessary to replenish water together with the solvent or alone. If the water content is too low, the adsorption rate of coenzyme Q to the hydrophobic adsorbent will be reduced, and therefore a large amount of adsorbent will be required per coenzyme Q to be extracted. Furthermore, if the water content is too high, the solubility of coenzyme Q in the solvent will be too low, resulting in a decrease in extraction rate.

The extraction temperature and time are not particularly limited, but the extraction temperature is preferably room temperature or room temperature to 50°C.

In addition, the extraction time varies depending on the type of extractant, type of raw material, condition, temperature, etc., but it is usually 20 to 30 minutes.
It may take from a minute to a night.

Contact between the coenzyme Q-containing material and the solvent is usually carried out by stirring an extraction mixture containing the coenzyme Q-containing material and the solvent.

5- To separate the extract from the extraction mixture after being stirred in this manner, conventional solid-liquid separation means such as centrifugation, sedimentation, and filtration are employed. In addition, since microbial cells are usually aggregated in the presence of a hydrophilic solvent, they are easily separated by sedimentation or filtered, and an extract can be easily obtained continuously.

The extract is then brought into contact with a hydrophobic adsorbent, and coenzyme Q is adsorbed onto the adsorbent.

Hydrophobic adsorbents are not particularly limited, but include chemically bonded silica gel with minimal residual silanol groups or porous synthetic resins such as commercially available Amberlite (trade name) and Hivolas (trade name). etc. are preferably used.

For this adsorption, these adsorbents are usually packed in a column, for example, and the extract is allowed to flow down the column. Alternatively, the extract and the adsorbent may be mixed and stirred or shaken.

The former is preferred industrially. In this way, coenzyme Q is adsorbed onto the adsorbent, and this adsorbent is separated from the adsorption residue.

Although the extract is usually subjected to the adsorption treatment as it is, it is not prohibited to add or remove water or a solvent as long as the water content does not fall within the range of the extraction.

The adsorption temperature is usually the same as or slightly higher than the extraction temperature, for example, about 5 to 10°C higher. If the adsorption temperature is lower than the extraction temperature, oily substances may precipitate and solidify, and the surface of the forebrain adhesive may be coated, causing column clogging.

In the present invention, a series of extraction-adsorption operations must be performed multiple times on the same coenzyme Q-containing material, but it is usually repeated two or more times, preferably about 4 to 10 times. Note that the extractant used each time may be updated each time, and the adsorption residual liquid obtained in the previous time may be recycled and reused, but from an industrial perspective, later application is preferable.

After adsorption, coenzyme Q is desorbed and eluted from the adsorbent using an organic solvent, and coenzyme Q is recovered and purified from this eluate. Although desorption may be carried out each time the above-mentioned adsorption is performed, it is performed after the adsorbent is almost saturated with coenzyme Q by several adsorptions. The organic solvent used for this desorption may be any organic solvent as long as it dissolves coenzyme Q well, and hydrophobic organic solvents such as petroleum ether and →-hexane can also be used, but the hydrophilic solvent used for extraction It is preferable to use the same type of solvent as it does not increase the number of types of solvent. In this case, it is preferable to use a solvent for desorption with a lower water content because the amount of solvent required for desorption can be reduced.

If necessary, by bringing a water-containing hydrophilic solvent into contact with the adsorbent prior to desorption, some of the adsorbed impurities can be desorbed, and the purity of the coenzyme Q elution fraction can be improved. can be improved.

Although the adsorption residue obtained by adsorption and which does not substantially contain coenzyme Q can be discharged outside the system, it is preferable to reuse it as an extractant in the extraction process.

In the adsorption according to the present invention, an extract with a relatively high water content is used as an adsorption raw material during adsorption, and a solvent with a relatively low water content is used during desorption. The adsorption capacity increases, and coenzyme Q and impurities are substantially completely desorbed by desorption, and after desorption, the adsorption capacity of coenzyme Q is substantially completely recovered, which is 1.
It does not get clogged and can be used repeatedly over a long period of time.

However, in the present invention, there is no need to separate the adsorbent from the solid extraction raw material and extraction residue, and the recovery rate of coenzyme Q is high, and the period of use of the adsorbent is long, and the resulting coenzyme High purity.

The present invention will be explained in more detail below with reference to Examples.

However, the present invention is not limited thereto.

Example 1 Protaminobacter cultured using methanol as a carbon source
600 g of bacterial cell concentrate (coenzyme Q+) was obtained by centrifugation from the culture solution 201 of bacteria of the genus 9-.

216 mg) was obtained. To this, 1,500 ml of 2 wt % aqueous acetone was added, and a 2-volume Erlenmeyer flask was used as an extraction tank, and the mixture was stirred at 35°C (the water content of the extraction mixture was 31.5
%). This extraction mixture was suction-filtered to obtain an extract, while the extraction residue remained in the extraction tank. This extract solution -q% was filled with 3 nmJ of porous synthetic resin Diaion Hp-211 (Mitsubishi Kasei Corporation) as an adhesive and allowed to flow down a column kept at 35°C to allow the adsorbent to adsorb coenzyme QIO. The extraction residual liquid discharged from the column was recycled to the extraction tank and reused. Extraction-adsorption-recycling continued in this manner for 17 hours. The amount of adsorption residue recycled during this period corresponded to the amount required to repeat the series of extraction-adsorption-recycling operations about 5 times.

Next, the coenzyme Q++ adsorbed on the adsorbent was desorbed and eluted by flowing 1 ooml of 2wt rich aqueous acetone. Coenzyme Q+a in the eluate was 210Tn9. This corresponds to a recovery rate of 97%.

10- Comparative Example 1, Bacterial cell concentrate 600 obtained in the same manner as Example 1, ji
Add 1500 TLl of 2 wt% aqueous acetone to
The mixture was stirred and extracted at ℃ for 17 hours. After completion of the extraction, the extract obtained by centrifugation was added to Diaion Hp-20, coenzyme Q, and the like in Example 1. After adsorbing
Elution was carried out in the same manner as in Example 1 using 2 wt % aqueous acetone.

In this case, the coenzyme Q+o in the eluate was 98■.

This corresponded to a recovery rate of 45 cm, which was an extremely low value.

Comparative Example 2 To 600 g of the bacterial cell concentrate obtained in the same manner as in Example 1,
wt hydrated acetone 1500ml and Diaio 7 Hp
-2030n11 was added together and stirred at 35°C for 17 hours. This extraction mixture was passed through a 100-mesh sieve to remove Diamond Ion T (p-20), but the bacterial cells were in an agglomerated state.
The residue remained on the sieve, so it could not be sieved as it was. Therefore, approximately 6 liters of water was added to this extraction mixture to break up the aggregation of the bacterial cells to some extent, and then sieved. However, some bacterial cells still remained on the sieve, so the sieve was immersed in a water tank and shaken. Finally, only the bacterial cells were allowed to pass through, and Diamond Ion Hp-20
I was able to sift through the

After filling the column with this Diamond Ion Hp-20,
Adsorb coenzyme Q with 2% aqueous acetone. was eluted.

Coenzyme Q+6 in the eluate in this case was 174. This corresponds to a recovery rate of about 81%.

Note that when the amount of Diaion Hp-20 used was 60 ml, which was twice the above amount, the recovery rate was about 90 ml.

Example 2 A culture solution 2) of bacteria of the genus Xanthobacter cultured as a methanol carbon source was centrifuged, and 300 g of bacterial cell concentrate (
Coenzyme Q+O content 108 ■) was obtained, and to this was added 12 wt% hydrated isopropanol (approximately equivalent to the composition with water) 75:O, and 35
It was stirred at ℃ (the water content of the extraction mixture at this time was 375
h) Extraction was performed using the same # as in Example 1 except for the following. The obtained extract was subjected to x
Coenzyme Q1° was adsorbed by flowing down a column filled with 5 ml and kept at 37°C. The adsorption residue that flowed out of the column was recycled to the extraction tank. This series of extraction-adsorption-recycling operations was continued for 4 hours. During this period, the amount of liquid recycled was equivalent to repeating the series of extraction-adsorption-recycling operations about 5 times. The coenzyme Qll+ adsorbed on the column was eluted by flowing down 100ml of 12 wt% aqueous isopropanol. Coenzyme Q1° in the eluate was 104 mg. This is a recovery rate of 96
Corresponds to

Comparative Example 3: Extraction was carried out in the same manner as in Example 2 for 4 hours, except that the adsorption residual liquid was not recycled, and after the extraction was completed, the extract obtained by centrifugation was adsorbed in the same manner as in Example 2. I took it off. The amount of coenzyme Qra in the obtained eluate was 65. This corresponded to a collection rate of only 60.

Comparative Example 4゜1 for 300g of bacterial cell concentrate obtained in the same manner as in Example 2.
2wt water-containing ingropanol 750d and 15ml of YMC gel ODS were added and extracted at 35°C for 4 hours.

MMC gel ODS was separated and adsorbed in the same manner as in Comparative Example 2, except that about 3 g of water was further added to this extraction mixture.
After desorption, coenzyme Q+a was eluted.

In this case, the amount of coenzyme Q+a in the eluate was 91.

This corresponds to a recovery rate of 84 cm.

Example 3 Prota mixed solvent 4 obtained by culture using methanol as a carbon source
An extract was obtained in the same manner as in Example 1 except that oamJ was added and stirred at 40°C, and this extract was adsorbed and desorbed in the same manner as in Example 2 to adsorb 14-coenzyme Qlo, and The adsorption residual liquid was recycled to the extraction tank. Extraction-adsorption-recycling continued for 3 hours.

During this period, the volume of the recycled adsorption residual liquid was measured, and the extraction-adsorption-recycling operation was repeated about 8 times.

Coenzyme Q, adsorbed on an adsorbent. was desorbed and eluted by flowing 15 nmJ of 2wt rich aqueous acetone. The amount of coenzyme Q+a in the eluate was 195m9. This is a recovery rate of 985
Corresponds to

Coenzyme Q and Coenzyme Q were prepared in the same manner as in Example 3 except for +-3-II and IA. was obtained only 96~. This was a recovery rate of only 49%.

Patent applicant: Mitsubishi Gas Chemical Co., Ltd. Representative Kazuyoshi Nagano 15- 505-

Claims (1)

[Claims] In the method for producing coenzyme Q from a coenzyme Q-containing material, an extraction treatment in which a hydrophilic solvent and a coenzyme Q-containing material are brought into contact in the presence of water, and a coenzyme Q extract obtained by the extraction treatment. A method for producing coenzyme Q, which comprises repeating an adsorption treatment in which coenzyme Q is adsorbed onto a hydrophobic adsorbent, and reusing the adsorption residual liquid obtained in the adsorption treatment as an extractant, if desired.