JP2016124998A - Algae fat extraction method, and ultrasonic processor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an algae fat extraction method which enables high-quality algae fat to be easily obtained in high yield, and an ultrasonic processor.SOLUTION: An algae fat extraction method 1) for extracting algae fat contained in alga is provided that comprises steps of: dissolving oxygen in an algae-containing liquid containing the algae; and obtaining an algae-crushed liquid by crushing the algae by providing the algae-containing liquid containing the algae, in which oxygen is dissolved, with ultrasonic waves. An ultrasonic processor 2) is also provided that comprises: a processing tank for storing the algae-containing liquid containing the algae; and ultrasonic wave generating means for obtaining the liquid containing algae-crushed liquid by providing the algae-containing liquid in the processing tank with ultrasonic waves.SELECTED DRAWING: None

Description

  The present invention relates to a method for extracting algal fats and oils and an ultrasonic treatment apparatus.

  In recent years, fuel production using algae has been proposed as an alternative fuel for fossil fuels. Microalgae have a high light collecting ability and carbonic acid fixing ability. Therefore, in the fuel production using the microalgae, the oil and fat productivity per unit area is high, and is attracting attention as an excellent oil and fat production technology.

However, in the existing method, when extracting fats and oils from algae, refining is performed in a plurality of steps of two or more stages, and the increase in processing costs is a problem.
As an example of a conventional method for extracting TAG (triacylglycerol) from algae, first, the algae culture solution in which algae is cultured is allowed to stand, and then solid-liquid separation is performed to concentrate or dry the algae (first stage) ). Next, the obtained algae concentrate is subjected to physical extraction using a homogenizer or the like, or chemical extraction such as solvent extraction to extract oils and fats from the algae concentrate (second stage). ). However, since the fats and oils obtained by the conventional method have a low degree of purification of TAG, it is necessary to further perform a column separation operation or the like (third stage) (Non-Patent Documents 1 and 2).
At present, the oil refining cost accounts for a significant part of the biodiesel production cost, which is one of the obstacles to the practical application of biodiesel fuel. For this reason, the development of a new method for extracting algal fats and oils has been demanded.

Hiroshi Uno “Microalgae as Biomass Resources”, Mitsui Strategic Research Institute Report, Mitsui Strategic Research Institute, December 5, 2011, Internet <URL: http://mitsui.mgssi.com/issues/report/list_report11. php> Mitsufumi Matsumoto “Challenge to develop oil production technology for bio raw materials and fuels using microalgae”, Journal of Environmental Biotechnology, 2012, Vol. 12, No. 1, pages 9-14

  The present invention has been made in view of the above circumstances, and it is an object of the present invention to provide a method for extracting algal fats and oils and an ultrasonic treatment device that can easily obtain high-quality algal fats and oils in a high yield. To do.

  As a result of diligent research to solve the above problems, the present inventors have found that the above problems can be solved by crushing the algae by applying ultrasonic waves to the algae-containing liquid in which oxygen is dissolved, thereby completing the present invention. I let you.

  That is, the present invention provides an algal fat extraction method and an ultrasonic treatment apparatus having the following characteristics.

(1) A method for extracting algal fats and oils that extracts algal fats and oils contained in algae,
Dissolving oxygen in the algae-containing liquid containing the algae;
Crushing the algae by applying ultrasonic waves to the algae-containing liquid in which oxygen is dissolved to obtain an algae crushing liquid;
A method for extracting algal fats and oils, comprising:
(2) The method for extracting algal fats and oils according to (1), further including a step of recovering the algal fats and oils from the algal crushing liquid using an organic solvent.
(3) The method for extracting algal fats and oils according to (1) or (2) above, wherein the ultrasonic frequency is 500 kHz or more.
(4) The method for extracting algal fats and oils according to any one of (1) to (3), wherein the ultrasonic frequency is in a range of 1 MHz to less than 2.5 MHz.
(5) The method for extracting algal fats and oils according to any one of (1) to (4), wherein a frequency of the ultrasonic wave is in a range of 1.3 MHz to 2 MHz.
(6) The algal fat or oil according to any one of (1) to (5), wherein oxygen is added to the algae-containing liquid so that a singlet oxygen concentration in the algae-containing liquid is 20 μM to 270 μM. Extraction method.
(7) Extraction of algal fats and oils according to any one of (1) to (6), wherein oxygen is added to the algae-containing liquid so that the hydroxyl radical concentration in the algae-containing liquid is 10 μM to 100 μM. Method.
(8) The method for extracting algal fat according to any one of (1) to (7), wherein the addition of oxygen to the algae-containing liquid feeds an oxygen-containing gas into the algae-containing liquid by bubbling.
(9) The method for extracting algal fats and oils according to any one of (1) to (8), including a culture step of culturing algae to obtain the algae-containing liquid.
(10) a treatment tank for storing algae-containing liquid containing algae;
An ultrasonic processing apparatus comprising: ultrasonic generation means for applying ultrasonic waves to the algae-containing liquid in the treatment tank to crush the algae and obtain an algae crushing liquid.
(11) The ultrasonic processing apparatus according to (10), wherein a frequency of the ultrasonic wave is selected in a range of 500 kHz or more.
(12) The ultrasonic processing apparatus according to (10) or (11), wherein a frequency of the ultrasonic wave is selected in a range of 1 MHz or more and less than 2.5 MHz.
(13) The ultrasonic processing apparatus according to any one of (10) to (12), wherein a frequency of the ultrasonic wave is selected in a range of 1.3 MHz to 2 MHz.
(14) The ultrasonic processing apparatus according to any one of (10) to (13), further including a feeding pipe for feeding an oxygen-containing gas into the algae-containing liquid.
(15) A working liquid tank for storing a working liquid that propagates ultrasonic waves emitted from the ultrasonic wave generation means to the algae-containing liquid in the treatment tank,
The ultrasonic processing apparatus according to any one of (10) to (14), wherein the processing tank is disposed inside the working liquid tank.
(16) The ultrasonic processing apparatus according to any one of (10) to (15), further including an organic solvent storage tank that stores an organic solvent for recovering algal fat from the algal crushing liquid.

According to the method for extracting algal fats and oils of the present invention, high-quality algal fats and oils can be obtained in a high yield despite the treatment simplified as compared with the conventional method.
According to the ultrasonic processing apparatus of the present invention, it is possible to easily obtain a high-quality algal fat in a high yield.

It is a figure which shows an example of the chemical reaction considered to occur when an ultrasonic wave is given to water in which oxygen is dissolved. 1 is a diagram illustrating a schematic configuration of an ultrasonic processing apparatus according to an embodiment of the present invention. 1 is a diagram illustrating a schematic configuration of an ultrasonic processing apparatus according to an embodiment of the present invention. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution which was ultrasonicated for 10 seconds at 1.65 MHz. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution which was ultrasonicated for 20 seconds at 1.65 MHz. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution which was ultrasonicated for 30 seconds at 1.65 MHz. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution which was ultrasonically treated at 1.65 MHz for 60 seconds. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution before being ultrasonicated. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution which was ultrasonicated for 10 minutes at 1.65 MHz. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution which was ultrasonicated for 10 minutes at 47 kHz. In an Example, it is a photograph which shows the observation result of the algal cell contained in the culture solution which was ultrasonicated for 30 minutes at 47 kHz. It is a graph which shows the result of having measured the average particle diameter of the particle | grains contained in the algal culture solution after ultrasonication. In an Example, it is a graph which shows the difference of the TAG collection | recovery rate by the ultrasonic treatment time to an algal culture solution. In an Example, it is a graph which shows the result of having performed the ultrasonic treatment to the sample and comparing the singlet oxygen concentration and the hydroxyl radical concentration in the sample by the presence or absence of the bubbling treatment. In an Example, it is a photograph which shows the result of the thin layer chromatography with respect to the algal fat extracted by the extraction method of the algal fat according to the present invention. In an Example, it is a graph which shows the collection | recovery rate of TAG extracted from algae.

≪Method of extracting algal fats and oils≫
The method for extracting algal fats and oils of the present invention is an extraction method for algal fats and oils that extracts algal fats and oils contained in algae, wherein the step of dissolving oxygen in the algae-containing liquid containing the algae and the step of dissolving oxygen is performed. And applying an ultrasonic wave to the algae-containing liquid to crush the algae to obtain an algae crushing liquid.
In the present invention, “extracting algal fats and oils” means that the fats and oils present in the algal bodies are taken out of the algal bodies or released out of the algal bodies. With the extraction of the algal fat, the algae may be crushed.
Hereinafter, preferred embodiments of the present invention will be described.

  The method for extracting algal fats and oils of the present embodiment is a method for extracting algal fats and oils that extracts algae fats and oils contained in algae, comprising culturing algae to obtain the algae-containing liquid, and algae containing the algae. A step of dissolving oxygen in the containing solution, and a step of applying ultrasonic waves to the algae-containing solution in which oxygen is dissolved to crush the algae to obtain an algae crushing solution.

  Algae is not particularly limited, and almost all algae can be used. Microalgae is preferable, and unicellular green algae is preferable. As such algae, Nannochloropsis genus, Chlamydomonas genus, Botryococcus genus, Pseudocollistis genus, Feodactylum genus, Osterococcus genus, Cyanidiosizone genus, Klebsolmidium genus, Chlorochibus genus, Spirogilla genus, Examples include algae belonging to the genus Kara, Correokete, Chlorella, etc. Among them, Nannochloropsis, Chlamydomonas and Botryococcus are preferable. More specifically, Pseudochoristis ellipsoida, Botryococcus braunii, Cyanidizon melolae, and C. ) And the like.

  As algal fats and oils, in addition to triacylglycerol, glycolipids, phospholipids, fatty acids, and hydrocarbons may be used, and triacylglycerol is preferred.

The algae-containing liquid is a liquid containing algae to be extracted with fats and oils. The algae-containing liquid preferably contains water from the viewpoint of ultrasonic treatment described in detail later, and is preferably a mixture in which algae are contained or dispersed in water or an aqueous solution. As the mixture, an algae culture solution obtained by culturing algae and containing algae can be preferably used. Since the method for extracting algal fats and oils of the present embodiment can treat the algae culture solution without using the operation of solid-liquid separation of the culture solution, etc., it requires fewer operations than the conventional method. Oil extraction can be performed.
The algae-containing liquid may contain various nutrients such as salts, vitamins, and carbohydrates for culturing algae. Moreover, it is preferable that the algae-containing liquid contains a salt. When the salt is contained in the algae-containing liquid, it is possible to increase the production amount of hydroxyl radicals and singlet oxygen in the algae-containing liquid compared to the case where the algae-containing liquid does not contain salt. Become. Therefore, algal fats and oils can be extracted from algae with high efficiency. The salt is preferably a metal salt, and more preferably a metal salt of a transition metal.

  The method for culturing algae to obtain an algae-containing solution may be selected appropriately according to the type of algae to be cultured. In order to accumulate a large amount of fats and oils in algae, phosphorus ions may be reduced from a culture solution for culturing algae.

The method for extracting algal fats and oils of the present embodiment includes a step of dissolving oxygen in the algae-containing liquid containing the algae, crushing the algae by applying ultrasonic waves to the algae-containing liquid in which oxygen is dissolved, The process of obtaining.
The disruption of cells and tissues by ultrasonic waves has been conventionally performed by using an ultrasonic homogenizer as an ultrasonic disruption method.
However, as the inventors have studied in detail, the process of dissolving oxygen in the algae-containing liquid is performed, and ultrasonic waves are applied to the algae-containing liquid in which oxygen is dissolved, so that the yield and purity of fats and oils are improved. From the above, it was found that an algae crushing liquid in which algae were crushed in a very favorable state was obtained. The algal fats and oils released into the algal crushing liquid can be recovered from the algal crushing liquid with high yield and high purity. Hereinafter, each process will be described.

Although disrupting cells by ultrasonic waves is a conventional method, conventional ultrasonic cell disruption mainly involves physically disrupting cells.
When physically crushing algae, by applying ultrasonic waves to the algae-containing liquid, the ultrasonic waves propagate through the algae-containing liquid, and the algae contained in the liquid are crushed. In addition, when the ultrasonic wave has a relatively low frequency, physical cell disruption mainly occurs. Specifically, physical crushing of algae by generation of pressure by applying vibration to the liquid, and physical crushing of algae by cavitation by generation of fine bubbles can be mentioned.

  On the other hand, in the method for extracting algal fats and oils of the present embodiment, a step of dissolving oxygen in the algae-containing liquid, and an ultrasonic wave is applied to the algae-containing liquid in which oxygen is dissolved to crush the algae to obtain an algae crushing liquid. Since it has a process, the crushing of algae according to the present embodiment chemically crushes algae.

FIG. 1 is a diagram showing an example of a chemical reaction that is considered to occur when ultrasonic waves are applied to water in which oxygen is dissolved. As shown in FIG. 1, when ultrasonic waves are transmitted to water, the water is decomposed by the ultrasonic waves to generate hydroxyl radicals (OH.) And hydrogen atoms (sometimes called hydrogen radicals (H.)). .
Here, by performing a step of dissolving oxygen in the algae-containing liquid, hydroxyl radicals react with dissolved oxygen to generate hydrotrioxide radicals (HOOO.) As intermediates, and further the reaction proceeds to generate hydroxyl radicals. And singlet oxygen ( ¹ O ₂ ) is generated. The hydroxyl radicals produced here also react with dissolved oxygen, resulting in an increase in the concentration of singlet oxygen.

  By placing algae in a solution in which a reaction that occurs when ultrasonic waves are applied to water in which oxygen is dissolved, the algae reacts with the reaction product, and the algae is in a very favorable state from the viewpoint of oil extraction. Can be crushed. In particular, it is important that hydroxyl radicals, singlet oxygen, and algal cells coexist in one reaction system. Among the above reaction products, singlet oxygen and hydroxyl radical have a strong oxidative degradation action, and it is considered that algal cells are crushed by the oxidative degradation action.

  Thus, in this embodiment, by performing the step of dissolving oxygen in the algae-containing liquid, the amount of singlet oxygen and hydroxyl radicals generated in the algae-containing liquid is increased, and the algal cells are more effectively crushed. can do.

In this embodiment, it is preferable that the frequency of the ultrasonic wave given to the algae-containing liquid is 500 kHz or more. This is because when algae-containing liquid is irradiated with ultrasonic waves in a low frequency band of about 50 kHz to 500 kHz, the disruption of algae is mainly due to physical cell disruption by vibration rather than the above chemical cell disruption. It is. As the upper limit of the frequency of the ultrasonic wave applied to the algae-containing liquid, a frequency that can be technically achieved may be set as the upper limit. The ultrasonic frequency applied to the algae-containing liquid is preferably 500 kHz to 5 MHz, and more preferably 600 kHz to 5 MHz.
In addition, the inventors have a concentration nearly 100 times higher than when water is irradiated with ultrasonic waves in a low frequency band of 25 to 100 kHz when water is irradiated with ultrasonic waves of a high frequency band of 1.65 MHz. It was found that the hydroxyl radical (OH.) Of was generated. And as shown in the Example mentioned later, inventors give a high-quality oil and fat with high purity by giving an ultrasonic wave of a 1.65 MHz high frequency band to algae content liquid at high yield. I found that I can do it. Considering the above knowledge, the frequency of the ultrasonic wave applied to the algae-containing liquid is preferably 1 MHz or more and 5 MHz or less, preferably 1 MHz or more and 5 MHz or less, and more preferably 1.3 MHz or more and 5 MHz or less. preferable.
In addition, when the inventors irradiate water with ultrasonic waves in a high frequency band of 2.5 MHz, compared to when the water is irradiated with ultrasonic waves in a high frequency band of 1.65 MHz, the hydroxyl radical (OH.) The knowledge that the production | generation of is reduced was acquired. In consideration of these findings, the frequency of the ultrasonic wave applied to the algae-containing liquid is preferably less than 2.5 MHz, more preferably 500 kHz or more and less than 2.5 MHz, and 600 kHz or more and less than 2.5 MHz. More preferably, it is 1 MHz or more and less than 2.5 MHz, more preferably more than 1 MHz and less than 2.5 MHz, more preferably 1.3 MHz or more and less than 2.5 MHz, and 1.3 MHz The frequency is more preferably 2 MHz or less, particularly preferably 1.5 MHz or more and 2 MHz or less, and particularly preferably 1.5 MHz or more and 1.8 MHz or less.

  The step of dissolving oxygen in the algae-containing liquid is intended to increase the dissolved oxygen concentration in the algae-containing liquid when ultrasonic waves are applied to the algae-containing liquid. Therefore, after the step of dissolving oxygen in the algae-containing liquid containing the algae, the step of obtaining ultrasonic waves in the algae-containing liquid in which oxygen is dissolved to crush the algae to obtain an algae-crushing liquid Good. However, dissolved oxygen is considered to be released from the algae-containing liquid over time. Accordingly, simultaneously with the step of dissolving oxygen in the algae-containing liquid containing the algae, performing a step of crushing the algae by applying ultrasonic waves to the algae-containing liquid in which oxygen is dissolved to obtain an algal crushing solution. preferable. For example, it is preferable to set the step of obtaining an algae crushing solution by adding ultrasonic waves to the algae-containing solution containing algae and applying ultrasonic waves to the algae-containing solution to crush the algae.

Examples of the method for dissolving oxygen in the algae-containing liquid include a method of adding oxygen to the algae-containing liquid, and a generally known method can be employed. For example, a method of contacting an oxygen-containing gas with an algae-containing liquid, a method of stirring an algae-containing liquid with an agitator and mixing with the oxygen-containing gas, a method of blowing an oxygen-containing gas into the algae-containing liquid (bubbling), and oxygen in the algae-containing liquid Examples thereof include a method of adding a generator, and a method of mixing a liquid obtained by adding oxygen to an arbitrary liquid with an algae-containing liquid. The oxygen-containing gas may be composed only of oxygen, but air is preferable from the viewpoint of availability.
Among the above methods, oxygen can be easily and efficiently dissolved in the algae-containing liquid. Therefore, a method of sending an oxygen-containing gas to the algae-containing liquid by bubbling is more preferable, and a method of sending air to the algae-containing liquid by bubbling is preferable. Further preferred.

  Although it is considered that oxygen can be added to the algae-containing liquid by the vibration of the liquid or the generation of bubbles by the ultrasonic treatment itself, the step of dissolving oxygen in the algae-containing liquid in this embodiment is to crush the algae. Therefore, it shall be by a method other than the ultrasonic treatment.

The singlet oxygen concentration in the algae-containing liquid to which ultrasound is applied is preferably 20 μM or more, 25 μM or more, 30 μM or more, 40 μM or more, 50 μM or more, 60 μM or more, 70 μM or more, 80 μM or more, 90 μM or more, 100 μM or more. 20 μM to 270 μM, preferably 25 μM to 270 μM, more preferably 30 μM to 200 μM, and particularly preferably 80 μM to 200 μM. Note that M represents mol / m ³ .
By performing dissolution of oxygen in the algae-containing liquid and ultrasonic treatment so that the singlet oxygen concentration in the algae-containing liquid falls within the above range, the state of crushing the algae can be made more preferable.
Further, for example, the hydroxyl radical concentration in the algae-containing liquid to which ultrasound is applied is preferably 10 μM to 100 μM, preferably 40 μM to 100 μM, more preferably 25 μM to 60 μM, and 30 μM to 50 μM. More preferably it is.
By performing dissolution of oxygen in the algae-containing liquid and ultrasonic treatment so that the hydroxyl radical concentration in the algae-containing liquid falls within the above range, the state of crushing the algae can be made more preferable.

  In the present invention, the singlet oxygen concentration and the hydroxyl radical concentration in the algae-containing liquid are respectively measured by the singlet oxygen concentration and the hydroxyl radical concentration shown in Examples described later using an electron spin resonance apparatus (ESR). It shall be measured under conditions or compatible conditions. A commercially available electron spin resonance apparatus (ESR) can be used.

  The amount of oxygen added to the algae-containing liquid can be determined as appropriate, but as a guide, it is preferably 1 mL / min to 500 mL / min, and 20 mL / min to 200 mL / min with respect to 1 mL of the algae-containing liquid. Or less, preferably 50 mL / min or more and 150 mL / min or less.

  As an example, the time for which ultrasonic waves are applied to the algae-containing liquid is preferably 10 seconds to 30 minutes, more preferably 30 seconds to 10 minutes, and even more preferably 60 seconds to 10 minutes with respect to 1 mL of the algae-containing liquid. It is considered that high-purity fats and oils can be efficiently extracted by performing ultrasonic treatment for a time in the above range.

  The temperature of the algae-containing liquid to which ultrasonic waves are applied is not particularly limited, but as a guide, it may be about 0 ° C. to 80 ° C., and preferably about 10 ° C. to 40 ° C.

The density of algal cells contained in the algae-containing liquid to which ultrasound is applied is preferably 1 cell / mL to 2 × 10 ⁹ cells / mL, more preferably 1 cell / mL to 2 × 10 ⁷ cells / mL. preferable.

Many algae accumulate fats and oils in intracellular organelles called oil bodies. Therefore, in order to extract fats and oils from algae in a high yield, it is necessary to destroy the cell walls of algal cells and take out the oil body.
However, the algal cell disruption in the conventional extraction method of algal fat is a physical algal cell disruption, and the degree of disruption of the cell is probably too high, so that impurities such as pigments are mixed in addition to the target oil and fat component. That was the problem.

On the other hand, in this embodiment, preferably, by applying ultrasonic waves having a frequency of 600 kHz or higher to the algae-containing liquid, algal cells can be crushed in a state very suitable for extracting oil and fat, The fats and oils can be obtained in a high yield while greatly reducing the contamination of impurities in the extracted fats and oils. Moreover, by performing the step of dissolving oxygen in the algae-containing liquid, it is possible to obtain highly purified oil and fat with higher efficiency while keeping the purity of the extracted oil and fat at a high level. Therefore, according to the method for extracting algal fats and oils of the present embodiment, high-quality fats and oils can be obtained in a high yield while eliminating the purification of fats and oils components that have been conventionally required.
Thus, the algal fat extraction method of this embodiment is an epoch-making method that enables high-purity fat extraction in a short time and at low cost.

In the method for extracting algal fats and oils of this embodiment, the method may further include a step of recovering the algal fats and oils from the algal crushing liquid.
The method for collecting the algal fat from the algal crushing liquid is not particularly limited. For example, a suitable solvent is brought into contact with the algal crushing liquid, the alga crushing liquid is overlaid with the solvent, and the solvent is recovered. The method of doing is mentioned. The solvent is preferably an organic solvent, and the above method is a method generally known as a solvent extraction method. The organic solvent is preferably a linear, branched or cyclic saturated hydrocarbon having 6 to 10 carbon atoms, and more preferably hexane.
The solvent extraction method is preferable in that the oil and fat can be recovered only by overlaying the solvent on the algal crushing liquid, and the oil and oil recovery cost can be kept low. However, when the solvent extraction method is employed in the conventional oil and fat extraction method, impurities other than the target oil and fat component may be recovered in the solvent. On the other hand, in the method for extracting fats and oils of the present embodiment, an algae crushing liquid in which algae cells are crushed in a state that is very suitable for extracting fats and oils is obtained. High quality oils can be recovered.

≪Ultrasonic treatment equipment≫
The ultrasonic treatment apparatus of the present invention is a treatment tank for accumulating algae-containing liquid containing algae, and crushing the algae by applying ultrasonic waves to the algae-containing liquid in the treatment tank to obtain an algae crushing liquid Ultrasonic generation means.
The ultrasonic treatment apparatus of the present invention can be suitably used for carrying out the algal fat extraction method of the present invention. Hereinafter, preferred embodiments of the ultrasonic processing apparatus according to the present invention will be described with reference to the drawings, but the method for extracting algal fats and oils of the present invention is not limited to the method using the following ultrasonic processing apparatus.

(First embodiment)
The ultrasonic processing apparatus of the present embodiment has a processing tank for accumulating algae-containing liquid containing algae, and an action of propagating ultrasonic waves generated by the ultrasonic wave generation means to the algae-containing liquid in the processing tank. A working liquid tank for accumulating the liquid, ultrasonic generation means for crushing the algae by applying ultrasonic waves to the algae-containing liquid in the treatment tank, and obtaining an algae-crushing liquid; and the algae-containing liquid contains oxygen A feed pipe for feeding gas, and the treatment tank is disposed inside the working liquid tank.

  As shown in FIG. 2, the ultrasonic treatment apparatus 10 of this embodiment includes a treatment tank 1, a working liquid tank 2, an ultrasonic wave generating means 3, and a feed for feeding an oxygen-containing gas into the algae-containing liquid. A tube 4. The ultrasonic wave generating means 3 is disposed in the working fluid tank 2.

  The ultrasonic processing apparatus of this embodiment includes a feed pipe 4 and a pump 5 as oxygen supply means. The oxygen-containing gas supplied from the pump 5 is supplied to the processing tank 1 through the feed pipe 4. The processing tank 1 is a tank that stores the algae-containing liquid L1 to which ultrasonic waves are applied. The opening at the tip of the feed pipe 4 for ejecting the oxygen-containing gas is provided below the liquid surface of the algae-containing liquid L1. When the pump 5 is operated, the oxygen-containing gas is blown into the algae-containing liquid L1, and the dissolved oxygen concentration in the algae-containing liquid L1 is increased.

  When the ultrasonic wave generating means 3 emits an ultrasonic wave in a state where the working liquid L2 propagating ultrasonic waves is accumulated in the working liquid tank 2, the ultrasonic wave emitted from the ultrasonic wave generating means 3 is generated in the working liquid tank 2. Is transmitted to the algae-containing liquid L1 in the treatment tank 1 via the working liquid L2. The algae-containing liquid to which the ultrasonic wave is applied is crushed in the treatment tank 1 to form an algae crushing liquid.

The ultrasonic frequency of the ultrasonic wave generating means is preferably 500 kHz or more. This is because when the algae-containing liquid is irradiated with ultrasonic waves in a low frequency band of about 50 kHz to 500 kHz, the disruption of the algae is mainly due to physical cell disruption by vibration rather than the above chemical cell disruption. It is to become. As the upper limit of the ultrasonic frequency of the ultrasonic wave generation means, a frequency that can be technically achieved may be set as the upper limit value. The ultrasonic frequency of the ultrasonic wave generating means is preferably 500 kHz or more and 5 MHz or less, and more preferably 600 kHz or more and 5 MHz or less.
In addition, the inventors have a concentration nearly 100 times higher than when water is irradiated with ultrasonic waves in a low frequency band of 25 to 100 kHz when water is irradiated with ultrasonic waves of a high frequency band of 1.65 MHz. It was found that the hydroxyl radical (OH.) Of was generated. And as shown in the Example mentioned later, inventors give a high-quality oil and fat with high purity by giving an ultrasonic wave of a 1.65 MHz high frequency band to algae content liquid at high yield. I found that I can do it. Considering the above knowledge, the ultrasonic frequency of the ultrasonic wave generating means is preferably 1 MHz or more and 5 MHz or less, preferably 1 MHz or more and 5 MHz or less, more preferably 1.3 MHz or more and 5 MHz or less. preferable.
In addition, when the inventors irradiate water with ultrasonic waves in a high frequency band of 2.5 MHz, compared to when the water is irradiated with ultrasonic waves in a high frequency band of 1.65 MHz, the hydroxyl radical (OH.) The knowledge that the production | generation of is reduced was acquired. Considering these findings, the ultrasonic frequency of the ultrasonic wave generating means is preferably less than 2.5 MHz, more preferably 500 kHz or more and less than 2.5 MHz, and 600 kHz or more and less than 2.5 MHz. More preferably, it is 1 MHz or more and less than 2.5 MHz, more preferably more than 1 MHz and less than 2.5 MHz, more preferably 1.3 MHz or more and less than 2.5 MHz, and 1.3 MHz The frequency is more preferably 2 MHz or less, particularly preferably 1.5 MHz or more and 2 MHz or less, and particularly preferably 1.5 MHz or more and 1.8 MHz or less.

  The pump 5 that is a supply source of the oxygen-containing gas and the ultrasonic wave generating means 3 can be operated sequentially or simultaneously.

The ultrasonic wave generating means is, for example, an ultrasonic vibrator. The ultrasonic processing apparatus may further include a motor for driving the ultrasonic transducer.
In addition, in this embodiment, although the aspect in which the ultrasonic wave generation means 3 was arrange | positioned in the working fluid tank 2 was illustrated, for example, the wall surface of the working fluid tank 2 outside the working fluid tank 2 is the ultrasonic wave generating means 3 The ultrasonic wave generating means 3 may be embedded inside the wall of the working fluid tank 2.

Moreover, in this embodiment, although the ultrasonic wave generation means 3 illustrated the aspect arrange | positioned in the inside of the working fluid tank 2, as another aspect, the ultrasonic wave generation means 3 is arrange | positioned in the processing tank 1. FIG. May be. In this case, the ultrasonic wave emitted from the ultrasonic wave generating means is given to the algae-containing liquid without passing through the working liquid, and the working liquid tank is not an essential component.
As shown in this embodiment, the working fluid tank 2 is provided, and the ultrasonic wave generating means 3 is disposed in the working liquid tank 2 so that the ultrasonic wave generating means and the algae-containing liquid are directly connected. Ultrasonic processing can be performed without contact. By setting it as such a structure, since it is hard to receive to the influence of algae containing liquid with respect to an ultrasonic generation means, it can be set as the ultrasonic processing apparatus excellent in manageability, durability, etc.

  The feeding pipe 4 may be connected to the processing tank 1, and a communication port communicating with the feeding pipe 4 may be provided on the wall surface of the processing tank 1.

  According to this embodiment, the oxygen-containing gas is introduced into the algae-containing liquid by the oxygen supply means. When ultrasonic waves are applied to the algae-containing liquid in a state where the dissolved oxygen concentration is increased by the ultrasonic wave generating means, the algal cells can be more effectively crushed as described in the method for extracting algal fats and oils. And high quality algal fats and oils can be obtained in high yield.

(Second Embodiment)
As shown in FIG. 3, the ultrasonic processing apparatus 20 of the present embodiment further includes an organic solvent storage tank 6 and a pipe 7 for accumulating an organic solvent for recovering algal fats and oils from the algal crush liquid. It differs from the ultrasonic processing apparatus 10 shown in FIG. Other configurations are the same as those of the ultrasonic processing apparatus 10, and the description of the points common to the ultrasonic processing apparatus 10 of the first embodiment is omitted. For the same components, the ultrasonic processing apparatus 10 is used. The same symbols as used are used.

  The organic solvent stored in the organic solvent storage tank 6 is introduced into the processing tank 1 through the pipe 7. When the organic solvent is introduced from the organic solvent storage tank 6 into the processing tank 1 while the algal crushing liquid is stored in the processing tank 1, the fats and oils in the algal crushing liquid are recovered into the organic solvent.

  According to this embodiment, since the organic solvent can be introduced into the algal crushing liquid and there is no need to send the algae crushing liquid, the running cost of oil extraction can be reduced.

  EXAMPLES Next, although an Example is shown and this invention is demonstrated further in detail, this invention is not limited to a following example.

1. Culture of algae As algae, a wild strain of Chlamydomonas (C9 strain CC-408 mt ⁻ ) was used. In normal culture, a modified medium of liquid TAP medium was used, and swirling culture was performed at 23 ° C. under continuous light of 20 to 40 μE m ⁻² s ⁻¹ . In order to accumulate a large amount of fats and oils, phosphorus ions are removed from the modified medium of the liquid TAP medium. Hereinafter, the modified medium of the liquid TAP medium is referred to as TAP modified medium (-P). In the experiment, cells transferred to TAP-modified medium (-P) and cultured were used. The process of transferring is as follows. Chlamydomonas alga bodies in the logarithmic growth phase are collected by centrifugation (5 minutes, 1,800 × g), washed twice with TAP-modified medium (−P), resulting in a cell concentration of 1.0 × 10 ⁵ cells / mL The suspension was suspended in TAP-modified medium (-P) as described above, and swirled and cultured at 23 ° C. for 8 days under continuous light of 20 to 40 μE m ⁻² s ⁻¹ .

2. Ultrasonic treatment of algae culture solution As described above (1. algae culture), algae is cultured, and the cultured algae are TAP so that the number of cells becomes 1-2 × 10 ⁷ cells / mL. This was suspended in a modified medium (-P), which was used as an algae culture solution, and 1 mL was used for sonication. The ultrasonic treatment is performed using an ultrasonic treatment device (Spin Nano Device IV (manufactured by Nimo Co., Ltd.)) at an output of 36 V and 1.65 MHz for 10 seconds, 20 seconds, 30 seconds, or 60 seconds. Or ultrasonication (room temperature) for 10 minutes at an output of 48 V and 1.65 MHz. The ultrasonic treatment of the algae culture solution is the tank attached to the spin nano device, and water is put into a tank provided with an ultrasonic vibrator inside the tank, and the algae culture solution is put into the water stored in the tank. The test tube was submerged. In addition, as a control group, ultrasonic treatment (room temperature) was performed on an algae culture solution at an output of 47 kHz for 10 minutes or 30 minutes using an ultrasonic cleaning apparatus (BRANSONIC 3210, manufactured by Nippon Emerson). The test was carried out in a state where the test tube containing the algae culture solution was submerged in the water stored in the tank of the ultrasonic treatment apparatus.
Immediately after the ultrasonic treatment, observation was performed with an optical microscope.

4 to 11 show the observation results of the algal cells contained in the culture solution that was sonicated under each of the above conditions.
4 shows an output of 36 V at 1.65 MHz after sonication for 10 seconds, FIG. 5 shows an output of 36 V at 1.65 MHz for 20 seconds, and FIG. After sonication for 30 seconds at 65 MHz, FIG. 7 shows the state of the algal cells after sonication for 60 seconds at an output of 36 V and 1.65 MHz.
FIG. 8 shows the state of algal cells before sonication, and FIG. 9 shows the state of algal cells after sonication for 10 minutes at an output of 48 V and 1.65 MHz.
FIG. 10 shows the state of algal cells after sonication at 47 kHz for 10 minutes, and FIG. 11 shows the state of algal cells after sonication at 47 kHz for 30 minutes.

As shown in FIGS. 10 and 11, it can be seen that the cells are not crushed well in the 47 kHz ultrasonic treatment.
On the other hand, in the 1.65 MHz ultrasonic treatment, as shown in FIGS. 4 to 7, it is understood that the algal cells are crushed as the treatment time of the ultrasonic treatment is increased. Moreover, as FIG. 9 shows, it turns out that the algal cell was destroyed favorably by the ultrasonic treatment for 10 minutes.

3. Measurement of particle size of particles contained in algae culture solution after sonication As described above (1. algae culture), algae is cultured and the number of cells becomes 1-2 × 10 ⁷ cells / mL Algae bodies were prepared as described above, and 1 mL was used for ultrasonic crushing. The processing conditions were an output of 48 V and 1.65 MHz for 10 minutes. Samples before and after the treatment were appropriately diluted, and the particle size was measured with a laser diffraction particle size distribution measuring apparatus. SALD-2300 manufactured by Shimadzu Corporation was used for the laser diffraction particle size distribution measuring apparatus.

  The results are shown in FIG. The average particle size of the particles contained in the algal culture solution before sonication was about 10 μm, and the average particle size of the particles contained in the algal culture solution after sonication was about 1.3 μm. . From this, it was confirmed that the algal cells in the algae culture broth were destroyed by ultrasonication to become smaller particles.

4). Examination of ultrasonic treatment time As described in the above (1. Culture of algae), algae is cultured, and algal cells are prepared so that the number of cells is 1-2 × 10 ⁷ cells / mL. 1 mL was used for ultrasonic crushing. The ultrasonic treatment conditions were an output of 48 V, 1.65 MHz, and ultrasonic treatment was performed at an algal culture solution temperature of 50 ° C. for 5 minutes, 10 minutes, 20 minutes, or 30 minutes.
Each 1 mL of the algae culture solution was overlaid with 1 mL of hexane and allowed to stand overnight. Thereafter, the hexane layer was recovered, and thin layer chromatography (TLC Silica gel 60, 20 × 20 cm, Merck, product code 1.05721.0009, developing solvent composition was hexane: diethyl ether: acetic acid = 160: 40: 4 (vol / vol / vol)).
The content of the oil and fat triacylglycerol (TAG) was scraped from the plate and the content was measured.
TAG was subjected to methanolysis treatment using 15: 0 fatty acid as an internal standard sample. Specifically, in a glass test tube with a screw cap, 100 μl of 1 mM 15: 0 hexane solution (pentadecanic acid, Sigma-Aldrich, P-6125) and 350 μl of 5% (vol / vol) chloride in silica gel powder containing TAG A hydrogen methanol solution (Wako Pure Chemicals, 089-03971) was added and treated at 85 ° C. for 1 hour. After the methanolysis treatment, fatty acid methyl ester was recovered with hexane, dried with nitrogen gas, and then recovered with 60 μl of hexane, of which 3 μl was analyzed by gas chromatography. Gas chromatography (Shimadzu Corporation, GC-2014, column is HR-SS-10 (25 m × 0.25 mm ID) manufactured by Shinwa Kako Co., Ltd.), column temperature 180 ° C., vaporization chamber and detector 250 ° C., inlet pressure ( kPa) 68.2, column flow rate (ml / min) 0.53, split ratio 68.8, measurement time 15 minutes).
The TAG recovery rate in the hexane layer was determined by extracting total lipids from algal cells not subjected to ultrasonic treatment based on the Bligh and Dyer method (Can. J. Biochem. Phsiol. (1959) 37, 911). TAG was quantified using thin layer chromatography and gas chromatography in the same manner as described above, and the recovery rate was calculated with the value as 100%.

  The results are shown in FIG. From the graph shown in FIG. 13, it was found that the amount of collected oil was almost the maximum when the ultrasonic treatment was performed for about 10 minutes at the alga-containing liquid temperature of 50 ° C. under the above treatment conditions.

5). Measurement of Singlet Oxygen Concentration and Hydroxyl Radical The amount of generated hydroxyl radicals and singlet oxygen was measured by ultrasonic treatment and bubbling treatment in addition to ultrasonic treatment. The bubbling treatment was measured in both cases of oxygen bubbling treatment and air bubbling treatment.
Singlet oxygen concentration and hydroxyl radical concentration were quantified using an X-band electron spin resonance (ESR) apparatus (JES-FA-100, manufactured by JEOL Ltd.). For the measurement of the hydroxyl radical concentration, a mixed solution of 150 μL of TAP-modified medium (-P) and 50 μL of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) (1.2 M) was prepared. For measurement of singlet oxygen concentration, 750 μL of TAP-modified medium (−P) medium and 250 μL of 2,2,5,5-tetramethyl-3-pyrroline-3-carboxamide (TPC) (100 mM) mixed solution Prepared.
Each sample was sonicated at 48 V output, 1.65 MHz for 1 minute or 5 minutes, and the ESR spectrum was measured. In addition, a bubbling treatment in which oxygen or air was blown at 100 mL / min was performed from 5 minutes before the ultrasonic treatment of each sample until the time when the ultrasonic treatment was completed.
The ESR measurement conditions are: microwave output 1 mW, microwave frequency 9.428 GHz, sweep magnetic field 330.5 to 340.5 mT, modulation frequency 100 kHz, modulation width 0.1 mT, sweep time 2 minutes, time constant 0 JES-FA-100 manufactured by JEOL Ltd. was used as the ESR spectrometer for 1 second. Each radical concentration was calculated based on the radical concentration of 20 μM TEMPOL using the signal area of Mn ²⁺ derived from manganese oxide as an internal standard substance and the signal area derived from the sample.

The results are shown in FIG. FIG. 14A is a graph comparing the singlet oxygen concentrations in the sample when the sample is subjected only to ultrasonic treatment and when the sample is subjected to ultrasonic treatment and oxygen bubbling treatment. FIG. 14B is a graph comparing the hydroxyl radical concentration in the sample when the sample is subjected only to ultrasonic treatment and when the sample is subjected to ultrasonic treatment and oxygen bubbling treatment.
FIG. 14C is a graph comparing the singlet oxygen concentration in the sample when the sample is subjected to only ultrasonic treatment and when the sample is subjected to ultrasonic treatment and air bubbling treatment. FIG. 14D is a graph comparing the hydroxyl radical concentration in the sample when the sample was subjected to only ultrasonic treatment and when the sample was subjected to ultrasonic treatment and air bubbling treatment.
It was shown that the singlet oxygen concentration in the medium increased in both cases of the oxygen bubbling treatment and the air bubbling treatment.
The hydroxyl radical concentration was different from the singlet oxygen concentration, but this was due to the generation of hydroxyl radicals from the two processes of water decomposition and hydrotrioxide radical decomposition. This is thought to be due to the consumption of this reaction.

6). Ultrasonic treatment and bubbling treatment on algae culture (Example 1)
Algae was cultured as described above (1. Culture of algae). The cultured algal cells were suspended in TAP-modified medium (-P) so as to have a cell number of 1 to 2 × 10 ⁷ cells / mL, and used as an algal culture solution. 1 mL of this algae culture solution was used for ultrasonic treatment and bubbling treatment.
The algae culture was sonicated at 48 V output, 1.65 MHz for 10 minutes at room temperature. In addition, oxygen bubbling treatment was performed in which oxygen was blown into the algae culture solution at a rate of 100 mL / min from 5 minutes before the ultrasonic treatment to the end of the ultrasonic treatment.

(Example 2)
The algae culture solution was subjected to ultrasonic treatment and air bubbling treatment in the same manner as in Example 1 except that air was blown into the algae culture solution at 100 mL / min instead of oxygen.

(Comparative Examples 1-2)
Algal culture solution was obtained in the same manner as Example 1. The algae culture was neither sonicated nor bubbled.

(Comparative Examples 3-4)
After obtaining the algae culture solution in the same manner as in Example 1, the algae culture solution was sonicated in the same manner as in Example 1. The algae culture was not bubbled.

7). Lipid extraction and recovery 1 mL of algae culture solution subjected to ultrasonic treatment and air bubbling treatment in Example 2 and 1 mL of algae culture solution subjected to ultrasonic treatment in Comparative Example 4 were overlaid with 1 mL of hexane. And left it overnight. Thereafter, the hexane layer was recovered, and thin layer chromatography (TLC Silica gel 60, 20 × 20 cm, Merck, product code 1.05721.0009, developing solvent composition was hexane: diethyl ether: acetic acid = 160: 40: 4 (vol / vol / vol)).
The results are shown in FIG. In the plate on which the aqueous layer was spotted (FIG. 15B), separation of other polar lipids typified by pigment, diacylglycerol and membrane lipid was observed in addition to the spot of oil (triacylglycerol). On the other hand, in the plate on which the hexane layer is spotted (FIG. 15 (a)), only the spot of oil (triacylglycerol) is observed, and separation of other polar lipids typified by pigments, diacylglycerol and membrane lipids is hardly observed. I could not confirm.
From this, it became clear that the purity of the fats and oils collected in the hexane layer from the algae culture solution that had been subjected to ultrasonic treatment was very high. Moreover, it became clear that the purity of the fats and oils which were collect | recovered by the hexane layer from the algal culture liquid which passed through the ultrasonic treatment and the air bubbling process is also very high purity. These recovered fats and oils are considered to have such a high purity that it is not necessary to purify the fats and oils that are usually required in the conventional fat and oil extraction method.

8). Measurement of the amount of TAG The spot of fat and oil triacylglycerol (TAG) was scraped from the plate to measure the content.
TAG was subjected to methanolysis treatment using 15: 0 fatty acid as an internal standard sample. Specifically, in a glass test tube with a screw cap, 100 μl of 1 mM 15: 0 hexane solution (pentadecanic acid, Sigma-Aldrich, P-6125) and 350 μl of 5% (vol / vol) chloride in silica gel powder containing TAG A hydrogen methanol solution (Wako Pure Chemicals, 089-03971) was added and treated at 85 ° C. for 1 hour. After the methanolysis treatment, fatty acid methyl ester was recovered with hexane, dried with nitrogen gas, and then recovered with 60 μl of hexane, of which 3 μl was analyzed by gas chromatography. Gas chromatography (Shimadzu Corporation, GC-2014, column is HR-SS-10 (25 m × 0.25 mm ID) manufactured by Shinwa Kako Co., Ltd.), column temperature 180 ° C., vaporization chamber and detector 250 ° C., inlet pressure ( kPa) 68.2, column flow rate (ml / min) 0.53, split ratio 68.8, measurement time 15 minutes).
The TAG recovery rate in the hexane layer was determined by extracting total lipids from algal cells not subjected to ultrasonic treatment based on the Bligh and Dyer method (Can. J. Biochem. Phsiol. (1959) 37, 911). TAG was quantified using thin layer chromatography and gas chromatography in the same manner as described above, and the recovery rate was calculated with the value as 100%.

The results are shown in FIG. FIG. 16 (a) shows the algae culture solution subjected to the ultrasonic treatment and oxygen bubbling treatment in Example 1, the algae culture solution of Comparative Example 1, and the algae culture solution subjected to ultrasonic treatment in Comparative Example 3. It is a graph which shows the collection | recovery rate of the fats and oils collect | recovered from each to the hexane layer.
Compared with the recovery rate of TAG from the algae culture solution of Comparative Example 1 and Comparative Example 3, the recovery rate of TAG from the algae culture solution of Example 1 has increased dramatically, achieving a high oil and fat recovery rate. It was shown that it was possible.

FIG. 16 (b) shows the algae culture solution that has been subjected to ultrasonic treatment and air bubbling treatment in Example 2, the algae culture solution of Comparative Example 2, and the algae culture solution that has been subjected to ultrasonic treatment in Comparative Example 4. It is a graph which shows the collection | recovery rate of the fats and oils collect | recovered by each from the hexane layer.
Compared with the recovery rate of TAG from the algae culture solution of Comparative Example 2 and Comparative Example 4, the recovery rate of TAG from the algae culture solution of Example 2 has increased dramatically, achieving a high oil and fat recovery rate. It was shown that it was possible.
Therefore, it was shown that a high oil recovery rate could be achieved even by air bubbling simpler than oxygen bubbling.

  The configurations and combinations thereof in the embodiments described above are examples, and the addition, omission, replacement, and other modifications of the configurations can be made without departing from the spirit of the present invention. Further, the present invention is not limited by each embodiment, and is limited only by the scope of the claims.

DESCRIPTION OF SYMBOLS 10,20 ... Ultrasonic processing apparatus, 1 ... Processing tank, 2 ... Working liquid tank, 3 ... Ultrasonic wave generation means, 4 ... Feeding pipe, 5 ... Pump, 6 ... Organic solvent storage tank, 7 ... Piping

Claims (16)

An extraction method for algae fats and oils that extracts algae fats and oils contained in algae,
Dissolving oxygen in the algae-containing liquid containing the algae;
Crushing the algae by applying ultrasonic waves to the algae-containing liquid in which oxygen is dissolved to obtain an algae crushing liquid;
A method for extracting algal fats and oils, comprising:   The method for extracting algal fats and oils according to claim 1, further comprising a step of recovering the algal fats and oils from the algal crushing liquid using an organic solvent.   The method for extracting algal fat according to claim 1 or 2, wherein the ultrasonic frequency is 500 kHz or more.   The method for extracting algal fats and oils according to any one of claims 1 to 3, wherein the ultrasonic frequency is in a range of 1 MHz or more and less than 2.5 MHz.   The method for extracting algal fats and oils according to any one of claims 1 to 4, wherein a frequency of the ultrasonic wave is in a range of 1.3 MHz to 2 MHz.   The method for extracting algal fats and oils according to any one of claims 1 to 5, wherein oxygen is added to the algae-containing liquid so that a singlet oxygen concentration in the algae-containing liquid is 20 µM to 270 µM.   The method for extracting algal fats and oils according to any one of claims 1 to 6, wherein oxygen is added to the algae-containing liquid so that a hydroxyl radical concentration in the algae-containing liquid is 10 µM to 100 µM.   The method for extracting algal fats and oils according to any one of claims 1 to 7, wherein the addition of oxygen to the algae-containing liquid feeds oxygen-containing gas into the algae-containing liquid by bubbling.   The method for extracting algal fats and oils according to any one of claims 1 to 8, further comprising a culture step of culturing algae to obtain the algae-containing liquid. A treatment tank for storing algae-containing liquid containing algae;
An ultrasonic processing apparatus comprising: ultrasonic generation means for applying ultrasonic waves to the algae-containing liquid in the treatment tank to crush the algae and obtain an algae crushing liquid.   The ultrasonic processing apparatus according to claim 10, wherein a frequency of the ultrasonic wave is selected in a range of 500 kHz or more.   The ultrasonic processing apparatus according to claim 10 or 11, wherein a frequency of the ultrasonic wave is selected in a range of 1 MHz or more and less than 2.5 MHz.   The ultrasonic processing apparatus according to any one of claims 10 to 12, wherein a frequency of the ultrasonic wave is selected in a range of 1.3 MHz to 2 MHz.   The ultrasonic processing apparatus as described in any one of Claims 10-13 provided with the feeding tube for sending oxygen-containing gas into the said algae containing liquid. A working liquid tank for storing a working liquid that propagates ultrasonic waves emitted from the ultrasonic wave generation means to the algae-containing liquid in the treatment tank;
The ultrasonic processing apparatus according to any one of claims 10 to 14, wherein the processing tank is disposed inside the working liquid tank.   Furthermore, the ultrasonic processing apparatus as described in any one of Claims 10-15 provided with the organic-solvent storage tank which accumulate | stores the organic solvent for collect | recovering algal fats and oils from the said algal crushing liquid.