KR20180128567A - The Mass Production Method of Odontella Aurita - Google Patents

Abstract

The present invention relates to a mass production method of microalgae Odontella Aurita. The purpose of the present invention is to provide a mass production method of Odontella Aurita, which can mass-produce microalgae Odontella Aurita having a strong antioxidative activity and an antiaging effect at low costs for a short period of time by using a medium. The mass production method of the present invention comprises: a step (a) of adjusting a pH value of seawater to a pH value range of 7.5 to 8.0 by using a neutralizing agent; a step (b) of adding an F/2 medium to the seawater to create a culture medium; a step (c) of inoculating 1 to 2 L of microalgae Odontella Aurita and a 10 to 20 L seed culture solution into an incubator; a step (d) of dividing the 10 to 20 L culture solution that has been cultured for 10 to 14 days into 100 to 200 culture solutions, and culturing the divided 100 to 200 culture solutions for 3 to 4 months to perform a primary scale-up process; a step (e) of inoculating the 10 to 20 L culture solution into a 200 to 400 L seed culture solution and culturing the culture solution inoculated into the seed culture solution for 10 to 14 days to perform a secondary scale-up process and obtain a secondary scale-up process completed culture solution; and a step (f) of recovering 50% of the culture solution from the secondary scale-up process completed culture solution, filling the recovered culture solution in the culture medium, and continuously culturing the culture solution filled in the culture medium for 1 to 2 months. The culturing process in each of the steps comprises culturing the culture solution such that the culture solution has a cell density of 16×10^4 to 20×10^4 cells/mL while supplying air in a rate of 0.03 to 0.05 m^3/min at 15 to 20°C.

Description

[0001] The present invention relates to a method for mass production of a micro-alga Odontella aurita,

The present invention relates to a method for mass production of a microalga Odontella aurita, and more particularly, to a method for mass production of a microalga Odontella aurita having a strong antioxidant ability and an anti-aging effect at a low cost in a short period of time using a medium It is about the mass production method of the Odontella aurita.

Recently, research has been reported that the ingredients contained in seaweeds are beneficial to human health. In particular, the fuco-xanthin component has been reported to have an anti-obesity effect, which is a unique carotenoid found abundantly in brown algae (K. Miyashita, Lipid Technology, August / September 2009, Vol. 21, No. 8/9) .

The anti-obesity mechanism of fucosanthin induces the expression of uncoupling protein 1 (UCP 1) in white adipose tissue (WAT) mitochondria resulting in fatty acid oxidation and heat production in WAT, resulting in apoptosis of adipocytes (apoptosis) action.

UCP 1 is a key molecule in the anti-obesity effect. The expression of UCP 1 is known to be an important component of body energy expenditure, and obesity is likely to occur when UCP 1 causes dysfunction.

Experiments showing the relationship between fucosanthin and UCP 1 expression and results of the experiment are disclosed in the above-mentioned documents. According to this document, fucosanthin induces protein and mRNA expression of UCP 1 in WAT. It appears to be due to the structural characteristics of fucoxanthin, namely the additional hydroxy substituents and allene bonds on the fucosanthin metabolites fucozanthinol and the sidechain groups of amarouscantanine A. (H. Maeda, Molecular Medicine Reports 2 : 897-902, 2009; and K. Miyashita, Same).

Such fucoxanthin has been separated and purified from brown algae in the past, but its concentration is not so large and there is a problem in that it takes time and expense for separation and purification, and therefore, attempts have been made to separate fucosanthin from microalgae, particularly Odontella aurita.

However, due to the limitations of media and culture techniques, it takes a lot of time and money to obtain enough production volume to isolate fucosanthin in sufficient quantity.

There have been attempts to cultivate Odontella aurita using synthetic media at some laboratory levels, but no results have yet been reported.

SUMMARY OF THE INVENTION Accordingly, the present invention has been made in view of the above circumstances, and it is an object of the present invention to provide a microalgae Odontella aurita having a strong antioxidant ability and an anti-aging effect and capable of mass- To provide a mass production method of the Odontella aurita.

The technical problem of the present invention as described above is achieved by the following means.

(1) adjusting the pH of the seawater to 7.5 to 8.0 using a neutralizing agent;

(b) adding F / 2 medium to the seawater to form a culture medium;

(c) inoculating a microalgae Odontella Aurita 1-2 L seed culture into a 10-20 L incubator;

(d) dividing the 10 to 20 L culture medium cultured for 10 to 14 days into 100 to 200, culturing for 3 to 4 months, and performing primary scale-up;

(e) inoculating the 10 to 20 L culture solution into a 200 to 400 L incubator and culturing for 10 to 14 days to perform a secondary scale-up;

(f) recovering 50% from the culture liquid in which the secondary scale-up has been completed, filling the culture medium and continuously culturing for 1 to 2 months,

The culture in each of the above steps is carried out by supplying air at a rate of 0.03 m 3 / min to 0.05 at 15 to 20 ° C. and culturing the cells to a cell density of 16 × 10 ⁴ to 20 × 10 ⁴ cells / mL. Mass production method of Aurita.

(2) In the above (1)

(a) adjusting the pH of the seawater to 7.5 with a neutralizing agent;

(b) adding F / 2 medium to the seawater to form a culture medium;

(c) inoculating 20 L of a microalga Odontella Aurita 2 L seed culture;

(d) dividing the 20-L culture medium for 14 days into 200, culturing for 3 months, and performing primary scale-up;

(e) inoculating the 20 L culture solution into a 400 L culture and culturing for 14 days to perform secondary scale-up;

(f) collecting 50% from the culture liquid in which the secondary scale-up is completed, filling the culture medium, and continuously culturing for 1 month,

Wherein the culture in each of the above steps is carried out so as to have a cell density of 16 x 10 ⁴ cells / mL while supplying air at 0.03 m 3 / min at 18 ° C.

As described above, according to the present invention, it is possible to produce a small algae Odontella aurita having a strong antioxidant ability and an anti-aging effect in a large quantity of Odontella aurita which can be mass-produced in a short time at low cost by using a medium.

Figure 1 is a 20 L culture profile of Odontella aurita according to the present invention.
Figure 2 is a 400 L culture profile of Odontella aurita according to the present invention.
Figure 3 is a 400 L continuous culture profile of Odontella aurita according to the present invention.
4 shows the cell density of Odontella aurita according to the present invention by 400 L reaction group.

According to an aspect of the present invention, there is provided a method for mass-

(a) adjusting the pH of the seawater to 7.5 to 8.0 using a neutralizing agent;

(b) adding F / 2 medium to the seawater to form a culture medium;

(c) inoculating a microalgae Odontella Aurita 1-2 L seed culture into a 10-20 L incubator;

(d) dividing the 10 to 20 L culture medium cultured for 10 to 14 days into 100 to 200 and culturing for 3 to 4 months;

(e) inoculating the 10 to 20 L culture medium into a 200 to 400 L incubator and continuously culturing the same for 1 to 2 months,

The culture in each of the above steps is characterized in that air is supplied at a rate of 0.03 to 0.05 m 3 / min at 15 to 20 ° C while culturing to a cell density of 16 × 10 ⁴ to 20 × 10 ⁴ cells / ml.

Hereinafter, the contents of the present invention will be described in more detail.

In the present invention, the culture medium of the microalga Odontella Aurita is obtained by filtering seawater through a filter device to remove impurities, and then neutralizing the pH to 7.5 to 8.0 using a neutralizing agent (Na ₂ S ₂ O ₃ ) And then adding thereto a known amount of F / 2 medium according to the conventional method. Each of the components constituting the F / 2 medium enhances the growth of microalgae. It is currently marketed in Korea through various sales companies. In the present invention, modified F / 2 medium supplemented with other additives can be used Of course.

Preferably, glycerol and / or organic acid may be added to the culture solution to increase the culture rate of the microalga Odontella Aurita. Examples of the organic acid include succinic acid, tartaric acid and citric acid.

The addition amount of glycerol is in the range of 100 to 200 g / ton, and when it is added at a rate of 100 g / ton, the effect of increasing the culture rate of the microalga Odontella Aurita is difficult to expect, , It is not preferable because the economical efficiency may be lowered or the growth may be adversely affected.

In the present invention, the organic acid is added to accelerate the micro-alga Odontella Aurita culture rate when used with glycerol, and it is sufficient to add the organic acid in a proportion of 100 to 200 g / ton of total organic acid .

In the present invention, micro-alga Odontella Aurita is inoculated into the culture medium prepared as described above and cultured in 1 to 2 L seed culture medium.

The 1 to 2 L seed culture obtained as described above was inoculated again into 10 to 20 L incubator and incubated at 15 to 20 ° C for 10 to 14 days at a rate of 0.03 to 0.05 m 3 / min to perform primary scale-up do.

Next, the 10-20 L culture medium is divided again into 100 ~ 200 cells, and cultured for 3 to 4 months under the same conditions to prepare a seed of 200 ~ 400 L culture medium.

The prepared 10-20 L culture medium was inoculated into 100-200 of 200-400 L incubators and cultured at 15-20 ° C for 10-14 days at 0.03-0.05 m 3 / .

After recovering 50% of the culture through the secondary scale-up, the culture medium is added again, and the cells are continuously cultured for 1 to 2 months. The microalga Odontella Aurita can be continuously and continuously cultured from the culture thus recovered, and the recovery process of the continuous culture liquid is as follows.

That is, in the present invention, a recovery line is provided at a height of 1/4 point of each incubator in order to recover the culture liquid, so that 1/2 of the culture liquid is allowed to escape. The recovered culture liquid is sent to a precipitation tank connected to the end of the recovery line Move.

The sedimentation tank containing the recovered culture medium is fed with a precipitant for sedimenting the microalga Odontella Aurita. As the precipitation agent used in the present invention, a polychlorinated aluminum flocculant is added, and the amount of the flocculant is preferably 0.4 to 0.5 mL per ton of the culture liquid.

When mixing the precipitant with the culture solution, mix the solution of the coagulant solution diluted to about 1000 times with water in small increments when the culture solution is collected in the precipitation tank so that the precipitant can be evenly mixed in the tank. The sedimentation time should be maintained for at least 12 hours to allow the sediment to settle down sufficiently on the bottom, and after the sedimentation, the culture fluid other than the float deposited on the floor should be drained through the discharge valve to maximize the sediment concentration.

Preferably, the operation of precipitating the microalgae, Odontella Aurita, as described above, is repeated two to three times after the general tap water is added and then the microalgae agglomerates are precipitated again. The salinity and the culture medium It is preferable to sufficiently remove the fine algae sediment.

In the next step, the precipitate of the most concentrated microalgae as described above is recovered and concentrated using centrifugation. Centrifuge operation is carried out by injecting the precipitate of microalgae at a speed of about 15000 ~ 18000 rpm at a speed of 500 ~ 600 L per hour through the lower part of the centrifuge, concentrating inside the centrifuge pawl, and then exiting only the clear culture liquid.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these embodiments are for the purpose of understanding the present invention and should not be construed as limiting the scope of the present invention.

[Example 1]

The seawater was adjusted to a pH of 7.5 with a neutralizing agent. F / 2 medium was added to the sea water according to the conventional method to prepare a culture medium, which was then cultured for 4 days to obtain a 2 L seed culture of Odontella Aurita. The 2L seed culture thus obtained was inoculated into a 20 L incubator and cultured at 15 캜 for 14 days with air being introduced at 0.03 m 3 / min (see the culture profile in FIG. 1). The 20L culture was further divided into 100 to 200 wells, maintained for 3 months, and prepared as seeds of a 400 L incubator. 20 L culture was inoculated into 200 L of a 400 L culture medium and cultured for 14 days (see the culture profile of FIG. 2).

After 14 days, 50% of the culture was recovered, and the culture medium was continuously added for one month. The culture profile according to the continuous culture was as shown in Fig. As described above, the cell density of the culture solution obtained as a result of continuous culture according to the embodiment of the present invention was measured as shown in FIG. 4, which was found to be 16 × 10 ⁴ / mL or more.

[Example 2]

The microalga Odontella Aurita was cultured in the same manner as in Example 1 except that the culture was added at a rate of 200 g / ton of glycerol.

[Example 3]

The microalga Odontella Aurita was cultured in the same manner as in Example 1, except that the culture medium was added at a ratio of 200 g / ton of glycerol and 200 g / ton of succinic acid.

[Example 4]

Odontella aurita was cultivated in the same manner as in Example 1, except that the culture medium was added at a ratio of 200 g / ton of glycerol, 100 g / ton of succinic acid and 100 g / ton of tartaric acid.

As can be seen from the above results, the present invention enables production of a high value-added food or cosmetic product at a low cost in a short period of time by using a microalgae Odontella Aurita in a culture medium and continuously culturing it It is expected that it will contribute greatly.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. It can be understood that

Claims (2)

(a) adjusting the pH of the seawater to 7.5 to 8.0 using a neutralizing agent;
(b) adding F / 2 medium to the seawater to form a culture medium;
(c) inoculating a microalgae Odontella Aurita 1-2 L seed culture into a 10-20 L incubator;
(d) dividing the 10 to 20 L culture medium cultured for 10 to 14 days into 100 to 200, culturing for 3 to 4 months, and performing primary scale-up;
(e) inoculating the 10 to 20 L culture solution into a 200 to 400 L incubator and culturing for 10 to 14 days to perform a secondary scale-up;
(f) recovering 50% from the culture liquid in which the secondary scale-up is completed, filling the culture medium, and continuously culturing for 1 to 2 months,
The culture in each of the above steps is carried out by supplying air at a rate of 0.03 m 3 / min to 0.05 at 15 to 20 ° C. and culturing the cells to a cell density of 16 × 10 ⁴ to 20 × 10 ⁴ cells / mL. Mass production method of Aurita. The method according to claim 1,
(a) adjusting the pH of the seawater to 7.5 with a neutralizing agent;
(b) adding F / 2 medium to the seawater to form a culture medium;
(c) inoculating 20 L of a microalga Odontella Aurita 2 L seed culture;
(d) dividing the 20-L culture medium for 14 days into 200, culturing for 3 months, and performing primary scale-up;
(e) inoculating the 20 L culture solution into a 400 L culture and culturing for 14 days to perform secondary scale-up;
(f) collecting 50% from the culture liquid in which the secondary scale-up is completed, filling the culture medium, and continuously culturing for 1 month,
Wherein the culture in each of the above steps is carried out so as to have a cell density of 16 x 10 ⁴ cells / mL while supplying air at 0.03 m 3 / min at 18 ° C.

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