JPS62146597A - Method for separating fat and oil component in mold - Google Patents

Abstract

PURPOSE:To collect high-purity fat and oil compnents produced by a mold, by adding a solvent to the dried mold, grinding the mold, bringing a supercritical fluid into contact with the mold and dissolving the fat and oils components in the mold in the supercritical fluid and the solvent. CONSTITUTION:A solvent is added to a mold which was dried to preferably <=7 wt.% water content, the mold is ground, fluidized and a supercritical fluid (fluid such as carbon dioxide, ethylene, ammonia, etc., forming a fluid in a supercritical state, having density close to that of liquid and a diffusion constant close to that of gas) is brought into contact with the mold so that fat and oils components in the mold are eluted in the solvent and the supercritical fluid. A filamentous fungus, especially Mortierella isabellina is preferable as the mold. Carbon dioxide is preferable as the superctirical fluid. A solvent suitable for uses of fats and oils selected from a lower alcohol, hydrocarbon ether and ketone is preferable as the solvent.

Description

[Detailed description of the invention] [Industrial use! ? ] This issue 1!1 relates to the separation power of fats and oils in bacterial cells, and will be explained in more detail in 1. This invention relates to a novel method for effectively separating oil and fat components produced by bacteria from bacteria.

[Prior art and its problems] In recent years, the supply H,+ from resources such as natural oils and fats has tended to be flat or depleted, and due to fluctuations in the harvest j11 due to changes in the weather, etc. Since the supply of oils and fats to the industry111 may fluctuate, a stable supply is desired in the food industry, cosmetics industry, pharmaceutical industry, etc.

Therefore, although there are methods to synthesize oils and fats that are the same as natural oils in a two-pronged manner, in industrial production, catalyst residue,
Impurities such as by-products are unavoidable, and how to remove these impurities is a major problem.

Also, 2 synthetic anti-J4. Since side reactions always occur, the problem in the synthesis of fats and oils is how to suppress side reactions and synthesize fats and oils efficiently.

In addition, as a new attempt, there seems to be a method of extracting the fats and oils in the bacterial cells with a solvent, but it requires the use of a single solvent.The water in the bacterial cells and the solvent form an emulsion. This may make it difficult to shunt the fats and oils from the solvent, and such methods are not always advantageous.
The industrial method is 1: difficult.

[Object of the Invention] This invention was made out of consideration of the above-mentioned (1).

In other words, the second feature of this invention is to create an oil and fat component based on a completely new concept, departing from the conventional chemical concept of reacting chemical substances with chemical substances to combine 1 nt oil component. The purpose of this article is to provide a method for extracting the oil and fat components produced by the bacteria, in more detail.
The purpose is to provide a method that is easy to use and can be used by humans with good yield.

[Step R for Achieving the Aforesaid Object] This invention has the following major points: Biosynthesis in bacterial cells is extremely specific, and specific substances can be biosynthesized chemically. , when the 1y1 body of 1 is cultivated, the special import substance of the large j11 is biosynthesized IIr moat 1.゛, -1 If it is possible to separate a specific component in a biosynthesized bacterial cell using a supercritical fluid, 1. -41f with high Xi line purity and high yield
It was completed in 611 years due to its ability to separate constant components.

In other words, the gist of the invention for achieving the 11th objective is as follows:
A solvent is added to the dry-cooked bacterial cells to crush and fluidize the bacterial cells, and then the bacterial cells are brought into contact with a supercritical fluid to convert oil and fat components in the bacterial cells into the supercritical fluid and the solvent. This is a method for separating oil and fat components in bacterial cells.

In this Part II, the oil and fat components are comprehensive, and include all the oil and fat components that affect the taste and are produced by the bacterial cells.

In general, oil and fat components, which are unrelated to microbial cells, are natural oils such as glycerin esters of fatty acids, oils and fats obtained from nature, purified by deodorizing, decolorizing, etc., and in some cases hydrogenated. Cosmetics b that can be added (11); Examples include oils and fats as ingredients, waxes containing L(m fatty acids and long-chain/hydric alcohol esters as one component), etc.

Natural oils and fats are classified into animal oils and fats and oils based on their origin, and these include mixtures of glycerin esters of various fatty acids shown below and 1. It's me. The fatty acids generally include saturated fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, montanic acid, and melisic acid, or Lindell's acid, Touhakuchu, and tuzunic acid. .

myristoleic acid, palmitoleic acid, petroselic acid, petroselysic acid, oleic acid, nilain,
Examples include fatty acids such as monoeno-M such as gontonic acid and erucic acid, trienoic acids such as niruhinic acid, linoleic acid, linoleic acid, linoleic acid, and linoleic acid.

In addition, waxes obtained from nature include the above-mentioned cholic acid and 1:
M# fatty acids, besides various esters with primary alcohols,
It is a complex mixture containing ML#alcohol, hydrogen crosslinking, polar fat, etc.

In this invention, all the substances and mixtures of glycerin esters of the various fatty acids that can be biosynthesized by the bacterial cells in the body are referred to as oil and fat components. 1) The types of oils and fats, waxes, and ingredients used are unknown.

Therefore, in this invention, if the type of bacterial cells can be determined, the oil and fat components obtained from them can also be determined.

To give an example, filamentous fungi such as Mortierella spp.
It is possible to obtain the oil and fat components used as ingredients.

Other examples of fungal cells include, for example, filamentous fungi such as Mortierella genus, Aspercillus genus, Talatosborium j.
Examples include the genus A.1core and the genus Limpus, and these fungal cells contain the same oil and fat components as those of the genus Mortierella and Izahelina described in III, although they differ in the amount of Ijj contained.

The advantage of Ir<n in this development 191 is that dried bacterial cells are used.

If undried bacterial cells are used as they are in the method of 91, the extraction process will take a lot of time, and water and oil and fat components will form an emulsion, making it impossible to efficiently separate the oil and fat components.

In this invention, if the moisture H,+ in the bacterial cell is 20 tonj11% or more, it may be called a dry bacterial cell, and furthermore, if the moisture H,- in the bacterial cell is less than I O+ +j-%. conveniently,
Further, the method of the present invention can be carried out even more conveniently if it is less than 7 tons and 111%.

For example, dry cooking of bacterial cells involves dehydrating the mixture of bacterial cells and water in a fermentation tank using a centrifugal machine, press filter, etc. to obtain a cake, flake, or other After washing the formed bacterial cell aggregate with water to remove the 1″X:a source in the bacterial cells, double tram drying,
The bacterial cells are dehydrated twice by spray drying, fluidized drying, etc. to bring the water content in the bacterial cells to within the above range.

The dried bacterial cells are crushed by adding a solvent to make them fluid.

The purpose of adding a solvent is to make crushing easier and to facilitate transportation, and also to improve the efficiency of dissolving oil and fat components into the supercritical fluid.

Therefore, jIi of the added solvent may be as low as 11: for example, the ratio of solvent/bacteria (1,000 r'+: Lt) may be 10 or more, preferably 4 to 0.5. The use of such a small amount of solvent is advantageous because the degree of emulsion formation between the oil and fat components and the solvent is small.

Various kinds of solvents can be used as the solvent, and there are no particular limitations. For example, lower alcohols, yJ aromatic hydrocarbons, Irtt ribbed carbon hydrocarbons, aliphatic ketones, etc. can be used. However, depending on the use of the 117 oil components separated by the method of the present invention, it is necessary to pay particular attention to points 1) and 5.

That is, if the oil IA component separated by the method of this invention is used in foods, cosmetics, medicines, etc., the solvents should be 1 Cosmetic Raw Materials 11 Food Additives 2, 11 Main Office It is preferable that it is product 11 listed in the Japanese version.

There are no particular limitations on the uses of the oil and fat components obtained by the method of the present invention, but if the uses are limited to foods, cosmetics, pharmaceuticals, etc., suitable solvents are, for example,
Examples include ethyl alcohol and n-hexano.

The fluidized bacterial cells first come into contact with a supercritical fluid.

A supercritical fluid is a fluid that forms a fluid in a critical state, such as ethylene, ammonia, or carbon oxide, and has a negative coefficient close to that of a liquid and a large diffusion coefficient close to that of a gaseous body. . Due to this characteristic, oil and fat components can be efficiently extracted from dried bacterial cells, and separation of oil and fat components from this supercritical fluid becomes easy.

Therefore, when a supercritical fluid is used in the method of this invention, 1; This is because six growing bacterial cells are brought into contact with a supercritical fluid.

The contact conditions can be determined depending on the type of supercritical fluid, such as a pressure and temperature of 1 min to maintain the tfl critical condition jE.

For example, if - oxidized carbon 7k is used.

The pressure is about 72.9 atmospheres 1^ or less, and the temperature is 31.0°C or less F1.
If ethylene is used, a pressure of 505 atmospheres or less, or a temperature of 9.9 degrees Celsius, can be recommended. Or even younger! - By recommending high temperatures,
If you use ammonia, the pressure should be 11
1.3 Qi Ha] or better J'; r +",'
, the power and temperature can be +32-3°C or higher.

However, considering the scale of I7, it is preferable to adopt carbon oxide 2+ as the supercritical 1 realm demon body and use it as the moon and force-claw. This is because it is a pollution hazard, and even if it remains there is no 4j, so it can be safely used for food products.

When dry bacterial cells to which a solvent has been added come into contact with a supercritical fluid,
The oil and fat components in the bacterial cells move to the solvent and the supercritical fluid.

Therefore, when a solvent and a supercritical fluid containing oil and fat components are taken out and the temperature and pressure are reduced, the oil and fat components that have been transferred to the solvent and supercritical fluid are diluted.

Next, an example of method V of the present invention will be further explained with reference to the drawings.

Figure 1 shows this 9. This is a separation device as an example of implementing the bright method υ.

In Fig. 1, dried bacterial cells are mixed with a solvent in a mixing tank 1 as shown in Figure 1.Next, the bacterial cells are crushed by stirring the mixture of the solvent and dried bacterial cells in a mill 2, and the mixture is mixed. Let be the fluid state yE.

Next, the microbial cells that have become fluid in the mill 2 are transferred to the extraction tank 3. In this extraction tank 3, supercritical state j! '; for example, carbon dioxide) and the crushed bacterial cells come into contact.

This extraction tank 3 is maintained at, for example, 40° C. and 200 atm, and the carbon oxide is in a supercritical state yE.

In the extraction tank 3, oil and fat components in the bacterial cells are transferred to a solvent and a supercritical fluid.

The bacterial cell residue from which the fat and oil components have been removed is appropriately extracted from the bottom of the extraction tank 3, for example.

The solvent and supercritical fluid containing oil and fat components are transferred to the first separation tank 5 via the transfer pipe, and this first separation tank R4fi
By lowering the pressure in the supercritical fluid and the solvent, the oil and fat components dissolved in the supercritical fluid and solvent are no longer dissolved and separated.
It accumulates at the bottom of the first separation tank 5. The separated oil and fat components are appropriately extracted from the bottom of the first separation tank 5.

- On the other hand, the supercritical fluid and solvent in the first separation tank 5 are transferred to the second separation tank 6 via a transfer pipe, and by reducing the pressure in this second transfer tank 6, the supercritical fluid is turned into scum. At the same time, the solvent is made into a liquid and dispensed into the bottom of the second separation tank 6.

The gasified carbon dioxide and molybdenum are supplied by a circulation pump (not shown).
is transferred from the second separation tank 6 by the pressure capo knob 4.
The fluid is pressurized and becomes a supercritical fluid, which is then pressurized into the extraction tank 3.

This method of development II (V) is not limited to the method shown in Fig. 1. For example, after extracting oil and fat components from bacterial cells in an extraction tank, the pressure in this extraction layer is reduced. Remove the bacterial extraction residue and oil and fat components that have accumulated at the bottom of the tube. 1. Pass the resulting mixture through a filter, centrifuge, and chopsticks to remove the oil and fat components. .

In addition, after the oil and fat components in the bacterial cells are transferred to the supercritical fluid and the solvent, the object of the present invention can be achieved by separating the oil and fat components by using a known appropriate chopstick.

[Example] Next, let's take a very small example of one person from IJ1.

(Examples 1 to 3) Inoculum of the filamentous fungus Mortierella genus Isabelina was obtained from the Chemical Branch Research Institute and was described in JP-A-59-205979 and JP-A-80-188391. It was more effective to do so.

The cultured viable bacterial cells were dehydrated using a centrifuge and washed with water. The obtained bacterial cells were dried in a double drum dryer to 150%
Dehydrated at °C. The water content +, + in the bacterial cells was adjusted by the drying time.

The moisture content in the bacterial cells was measured using a get infrared moisture meter. In addition, the oil and fat components in the bacterial cells are methanol-
A mixed solvent of chloroform was added to the bacterial cells, crushed in a ball mill, the solid content was removed by passing through one mouth, and methanol-
Chlorophort is Hy (a lot of oil and fat components! 11 is 'A')
Calculated by keeping it constant.

For example, in FIG. 1, in a 1ζ separation device, in a mixing tank, the oil and fat components of H and H shown in Table 1 and the dried bacterial cells (filamentous fungus Mortierella genus Isabelina) and the ITI shown in Table 1, '+j of ethanol (solvent) and stirred until the resulting mixture became fluid, thereby crushing the bacterial cells.

The fluid containing the bacterial cells was transferred to an extraction tank, and in the extraction tank, the bacterial cells and supercritical FE (dioxide) were brought into contact with the element. The temperature, pressure, and time during contact are listed in Table 1.

After the contact, the supercritical fluid IR element containing oil and fat components and the solvent are transferred to the first separation tank, and the temperature of the first separation tank is set to 5.
The oil and fat components were taken out from the bottom of the first separation tank by setting the temperature to 0°C and the pressure to 80 atm. Further, the solvent obtained from the second separation tank and the oil and fat components taken out from the first separation tank are mixed, the solvent is distilled off, and the remaining portions H and B are
The extracted oil and fat components were designated as H, +. The value obtained by dividing this ton jIt by 1,000 z of the oil and fat components in the prepared bacterial cells is calculated as the extraction value.
41 in Table 1.

In addition, the supercritical fluid and solvent after the oil and fat components have been separated are separated into solvent and gaseous G fat dioxide (5>lead) in a second separation tank, and the solvent is mixed with dry bacterial cells by +1 degree. When used, the raw material (gaseous dioxide) is pressurized to a supercritical state of 111 degrees and returned to the extraction tank for use in IIl Ikawa.

(Comparative Example 1.2) Using microbial cells whose water content i, j, and Separation of fat and oil components was carried out using Since the bacterial cells used in this comparative example have a high water content, it is difficult to extract oil and fat components, and the extraction efficiency 4t is very low.

(Comparative Example 3) Water-containing microbial cells similar to those in Example 2 were mixed with ethanol, crushed, and then subjected to solvent extraction by adding n-hexano. Note that n-hexane was added because ethanol hardly dissolves oil and fat components.

The extraction results are shown in Table 1.

(Comparative Example 4) Bacterial cell 1 containing moisture, +, ; similar to Example 2
00g, add 60-80 mesh glass peas to 150
In addition, after crushing, it is poured into a small extraction tank as shown in Figure 1, and extracted with supercritical fluid (-carbon oxide, also). [became.

The extraction conditions are shown in Table 1. Karas beads were used because it is difficult to disrupt the bacterial cells by themselves.

The extraction rate was lower than in Example 2.

“Invention of +IJ!!”J According to this issue 151, (1) Since dried bacterial cells are used, it takes a small amount to crush the bacterial cells.
It is only necessary to use a solvent of “l:”, (2)
By using the solvent in step 1 and drying the bacterial cells, it is possible to suppress the formation of emulsion between the moisture in the bacterial cells and the solvent, and therefore the extraction time for oil and fat components can be shortened accordingly. 3) Since it uses supercritical fluid, it is effective in 11: - (A) It can effectively extract oil and fat components!

(4) Since supercritical fluid is used, it is possible to efficiently separate one oil and fat component by applying 511° force, once turning, and lowering once. It is possible to provide the distribution of oil and fat components in the dried bacterial cells, which corresponds to the ratio 1:l.

Also, according to Equation 1!1, the fungal body produces dh-rich even number j and number 2 oil and fat components with H'71 purity T'i, so it is anti-1 ", ", like the A organism.・A: 1', +l containing pure ligature, qll 1lfj component can be separated into person j, -.

[Brief explanation of drawings]

FIG. 1 is a schematic 41 diagram illustrating an example of carrying out the method of the present invention. l...mixing tank, 2...mill, 3, ψ, extraction tank, 5◆
... 1st minute tank, 6... 2nd minute tank.

Claims (4)

[Claims] (1) Adding a solvent to the dried bacterial cells to crush and fluidize the bacterial cells, and then bringing the bacterial cells into contact with a supercritical fluid,
1. A method for separating oil and fat components in bacterial cells, which comprises including the oil and fat components in bacterial cells in the supercritical fluid and the solvent. (2) Claim 1, wherein the fungal body is a filamentous fungus.
The method for separating oil and fat components in bacterial cells as described in . (3) The method for separating oil and fat components in a fungus according to claim 1, wherein the fungus is a filamentous fungus of the genus Mortierella Isabelina. (4) The method for separating oil and fat components in bacterial cells according to any one of claims 1 to 3, wherein the supercritical fluid is carbon dioxide.