KR101413371B1 - A micro-algae culture method for increasing certain useful components using deep sea water. - Google Patents

Abstract

The present invention relates to a method for cultivating microalgae using LEDs (light-emitting diodes) having different wavelengths and cultivating pigments and components of Dunaliella cultured in seawater (deep sea water and deep sea water treated with ocean water) More specifically, a step of culturing the dunariella in seawater (sea deep seawater and deep seawater treated water), a step of culturing seawater (sea deep seawater and deep seawater treated seawater) as a culture medium, Culturing the microalgae (Dunaliella), comprising steps of culturing the microalgae, and culturing the microalgae (Dunaliella) comprising steps of increasing the concentration of the pigment components such as chlorophyll and beta-carotene in the body by applying stress to the dualiella.

Description

Technical Field [0001] The present invention relates to a micro-algae culture method for enhancing a specific useful ingredient using deep sea water.

The present invention relates to a method for producing microalgae, which comprises culturing a microalgae such as a dinaria in a deep sea water or a deep sea water treated with deep sea water and changing the environmental conditions such as light wavelength and salt concentration to produce a useful ingredient contained in microalgae such as dinaria To a culture method for promoting the growth.

Deep sea water is a clean sea water resource existing below 200m depth, and it is widely used in the production of deep sea water. Deep sea water (deep sea water concentrated in the ocean) generated in deep ocean water production process is higher in salinity than deep sea water, and contains minerals and nutrients. Such deep-seawater-treated water (deep-sea water concentrated in the ocean) is applied to food such as salt production, but since it is highly salted, there are not many cases where it is applied to bio-culture.

Dunaliella salina , a single-celled green alga, is known as a basophilic species with two flagella, and is resistant to high salt environments. D. salina is also one of the microalgae rich in beta-carotene and glycerol. It is known that production of β-carotene is more likely to be caused by environmental stress (intense radiation intensity, high saltiness, lack of nutrients and extreme temperatures).

On the other hand, in addition to minerals, our body needs a lot of nutrients such as protein, fat, carbohydrates and vitamins. Among them, antioxidants are currently attracting much attention in relation to human health. Our bodies are suffering from a variety of diseases such as aging, deterioration of cytoplasm and immunity due to the destructive action of harmful active oxygen and overproducing cells and tissues. Beta-carotene, lutein, chlorophyll, etc. have been known to be effective as functional materials that can prevent or alleviate these problems.

Beta-carotene is a carotenoid-based red pigment, which is found in red-colored carrots and tomatoes. Due to its unique color and efficacy, it has been added to foods and used. Its efficacy has been widely used as anticancer drugs, antioxidants related to skin aging, skin treatments, and antimicrobial agents. However, in spite of this usefulness, mass production is difficult and it is difficult to secure because it is limited in the quantity of included green leaf vegetables. The method of obtaining from the green leaf vegetables has a disadvantage that the amount of beta carotene is small and the harvest of the green leafy vegetables is dependent on the season and the climate change and consumes a long time for the cultivation. In order to overcome these problems, a method of chemically synthesizing has been sought, but it has been difficult to secure stability. Therefore, a method of mass-producing beta carotene and the like using microalgae has been sought.

However, the method of mass production of algae has a disadvantage in that it requires a wide space for culturing and it is required to continuously supply the microalgae with insufficient nutrients in the culture, and there is a problem in that the purity of the other algae can not be ensured by breeding together . In particular, farms constructed in open ponds are not able to maintain the highest salt tolerance because they are difficult to maintain high salt tolerance, and they are also ineffective in harvesting and separating cultured algae.

On the other hand, incandescent lamps have been used in the past as an adjustable light environment for the cultivation of living creatures instead of natural light, and the use of LED light has been increasing due to the development of LED light through fluorescent lamps. LEDs are long-lasting, low-power, environmentally friendly lighting that does not use discharge gas unlike fluorescent light. Although these characteristics are being used for the field cultivation of plants in recent years and LED is expected to be applied to cultivation of all kinds of land plants in the near future, it is still desired to increase useful components in the field of marine microalgae and algae cultivation There is no example of applying LED light to the LED.

The present invention proposes a microalgae culture method for improving the content of useful components such as beta-carotene by using deep sea water and LED as an economical method to overcome these problems.

The microalgae containing beta carotene, such as dinaria, spirulina and chlorella, are highly active in high salt environments. It is now confirmed that they are actively reproducing and propagating in the salt of Africa, the Dead Sea of Israel, and the natural salt field where the process of enrichment progressed to some extent. These high salt and high alkaline environments are the optimal conditions for their growth.

Main ingredients and properties of Duariella division main ingredient Functional Habitat environment (salinity,%) Duarielli
(Green algae) β-carotene, α-carotene, lutein, zeaxanthin, cryptosanthin, chlorophyll, omega-3 fatty acids, etc.  Protecting eyesight, preventing aging, improving immunity, ultraviolet protection of skin, antioxidant effect 3.3 to 15.0

Table 1 shows the major components and characteristics of the two larias, with the salinity of the habitat being 3.3-15.0%. Considering that the salinity concentration of the sea is usually 3.1-3.2%, it can be seen that it lives at a high salt concentration.

 Therefore, they are naturally concentrated in the evaporation, and have survived mainly in the highly salted soil. Of course, they can survive not only in this high salt environment but also in ordinary seas. Many nutrients are needed for them to grow. However, in the case of ordinary seawater, the supply of nutrients required by them is small, but when deep seawater is used, the nutrient-rich characteristics can be utilized relatively.

In other words, when the deep sea water is concentrated, the nutrients required for them are also enriched and a better growth environment is created. However, it is difficult to meet these conditions in a normal sea water environment and it is required to incur large costs.

Therefore, cultivating them using deep sea water (deep sea water concentrated in the ocean), which is a byproduct of the production of deep seawater, and using LED light to enhance the beneficial components such as beta carotene of birds, have.

Thus, in the present invention, a culture solution of D. salina is prepared by using deep sea water, concentrated water of deep sea water, surface water, NaCl, and the like to cultivate a microalgae, Duariella, and furthermore, This study is to investigate the growth and pigmentation of D. salina by establishing the environment, and to suggest a culture method in which betacarotin, which is a useful component of the cultivated daniariella, is promoted.

Korean Patent No. 10-06097360 relates to a microalgae culture apparatus and a microalgae culture method, which can realize a high productivity by realizing sufficient agitation of a culture solution, and also to attach microalgae to a wall of a culture vessel, A microalgae culture apparatus and method capable of preventing precipitation on the bottom surface and maintaining a high culture efficiency over a long period of time. Korean Patent No. 10-08970180 relates to a photobioreactor for culturing microalgae and a microalgae production apparatus having the same, and more particularly, to a microalgae production method using sunlight and light source irradiation efficiency to maximize the productivity of microalgae And a micro-algae production apparatus having the same. According to the present invention, microscopic algae are prevented from adhering to the inside of the reaction vessel, and the efficiency of light source irradiation is maximized by using a light guide plate or an OLED, thereby enhancing the productivity of microalgae, cultivating microalgae using sunlight, It is possible to provide a photobioreactor and a microalgae production apparatus for culturing microalgae that are operated without supplying microalgae. However, in order to achieve the object of the present invention, A wavelength value and the like for selectively increasing the wavelength of the light beam are not disclosed. In addition, Korean Patent Laid-Open Publication No. 10-2011-0094830 relates to a photobioreactor for micro-algae high-density culture, and a microalgae culture and harvesting method using the same. The micro-algae is manufactured in a closed structure, A microalgae culture reactor; A reactor cap detachably coupled to the top of the microalgae culture reactor; A culture fluid inlet pipe and an air discharge pipe inserted into the reactor stopper so as to communicate with the inside of the microalgae culture reactor; An air injection unit detachably coupled to a lower end of the microalgae culture reactor to supply air or combustion gas into the microalgae culture reactor; The present invention provides a microbiological culture bioreactor for microalgae culture, and a microalgae culture and harvesting method using the same. The present invention relates to a light incubator mainly used for the purpose of extracting oil by breeding microalgae in large quantities, and differs from the composition for enhancing useful components of the present invention.

The present invention is applicable to microalgae cultured by cultivating erythroblastic microalgae such as Duariella using sea water such as deep seawater and deep seawater treated water and irradiating light by wavelength using an LED light source under optical environmental conditions The present invention provides a culture method for promoting a component.

In order to solve the above problems, the seawater is concentrated to a high nutrient and high mineral state by RO (reverse osmosis membrane) apparatus and vacuum multi-stage evaporation concentrator or the like after pretreatment such as sterilization method or filtration method, The present invention relates to a method for efficiently culturing microalgae such as Duerella and the like at different wavelengths, and culturing the microalgae so as to contain a high content of useful substances such as beta-carotene.

More specifically, seawater is a deep sea water having a lot of clean and nutritious water, and the seawater for culturing the microalgae is prepared by directly using the seawater taken or by filtering the concentrated seawater, which is a byproduct remaining after desalination of the seawater, step; Adding a buffer such as ethylenediaminetetraacetic acid chelate (EDTA Chelate) or adding a culture medium containing a buffer to stabilize trace metals such as iron, copper and zinc contained in seawater.

Filtering, sterilizing and disinfecting seawater; Culturing a microalgae resistant to salt tolerance such as dunaliella, chlorella, and spirulina in an incubator capable of adjusting the culture environment such as light, air, temperature, and saline concentration; There is provided a microalgae culture method for enhancing useful components such as specific beta-carotene, chlorophyll, phycocyanin, lutein, zeaxanthin, vitamins or minerals, which is a target, while changing the culture environment and the culture period.

According to the present invention, it is possible to cultivate a dinariella containing useful substances such as enhanced beta carotene at a low cost from seawater such as deep ocean water rich in minerals and nutrients and deep sea water treated with ocean, Thus, it is possible to produce duariella with a variety of pigments and ingredients.

FIG. 1 is an overall process diagram according to a technique of culturing the change of pigment and component of duirialla according to wavelength.
2 is a photograph showing the production of concentrated water and permeated water using RO (reverse osmosis membrane).
Figure 3 is a view of an algal culture apparatus
FIG. 4 shows changes in the weight of microalgae according to wavelengths when deep seawater concentrated water, chelating agent and culture medium were used for each culture period.
FIG. 5 shows changes in pigmentation of microalgae according to wavelengths when deep seawater concentrated water, a chelating agent and a culture medium were used.
FIG. 6 shows changes in the effective components of microalgae according to wavelengths when using deep-sea concentrated water, a chelating agent and a culture medium.
FIG. 7 shows the results of Dunaliella The change of chlorophyll pigment by salinity of salina is shown.
Fig. 8 is a graph showing the distribution of Dunaliella The change of total carotenoid pigment by salinity of salina is shown.

In order to accomplish the object of the present invention, the present invention provides a method for producing microalgae, comprising the steps of: (a) preparing seawater for microalgae culture; (b) adding a buffering agent or a culture medium containing a buffer to stabilize trace metals such as iron, copper and zinc contained in seawater; (c) filtering, sterilizing and disinfecting seawater; (d) culturing a microalgae resistant to salt in an incubator capable of regulating the culture environment; (e) a microalgae culture method for enhancing a specific useful ingredient while changing a culture environment and a culture period or

(a) preparing seawater for microalgae culture; (b) filtering, sterilizing and disinfecting seawater; (c) culturing microbial flora resistant to salt in an incubator capable of regulating the culture environment; (d) a microalgae culture method is provided in which a specific beneficial ingredient to be augmented is changed while changing the culture environment and the culture period.

The seawater in step (a) may be obtained by directly using seawater taken as deep seawater having a clean and nutritious environment or by filtering concentrated seawater, which is a byproduct remaining after desalination of seawater, to remove suspended matters contained in seawater.

In the step (b), the buffer added may be ethylenediaminetetraacetic acid chelate (EDTA Chelate), and the sterilization and disinfection in the step (c) may be performed by UV or heat treatment, but not limited thereto , And the culture environment of step (d) can assume light, air, temperature, and salinity.

The salt tolerant microalgae of step (d) may be one or more algae selected from the group consisting of dunaliella, chlorella, and spirulina, and the specific useful components of step (e) include beta carotene, chlorophyll, , Zeaxanthin, vitamins, or minerals.

The incubation period of the microalgae may be culturing microalgae in seawater and culturing for 18-20 days. For the enhancement of picosanine, the LED light wavelength for improving specific useful components may be 420 ~ 480nm blue wavelength LED Light can be utilized. For the enhancement of beta-carotene, LED light having a red wavelength of 640 to 680 nm is used, and LED light having a yellow wavelength of 580 to 620 nm or a blue wavelength of 420 to 480 nm can be used for enhancing chlorophyll.

In the microalgae culture technique containing a useful substance such as beta-carotene according to an embodiment of the present invention, the microalgae may include various algae capable of growing in high saltiness without being limited to the double nariella. However, in the embodiment, the description will be centered on the Duariella.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

FIG. 1 shows a whole process diagram according to the technique of culturing the change of dye and component of dualiella according to wavelength, FIG. 2 shows a photograph of producing concentrated water and permeated water using RO (reverse osmosis membrane) It is the appearance of the algae culture device.

FIG. 4 shows the weight change of microalgae according to the wavelength when using the deep sea water concentrated water, the chelating agent and the culture medium for each culture period, and FIG. FIG. 6 shows changes in the effective components of microalgae according to wavelengths when deep seawater concentrated water, a chelating agent and a culture medium were used.

One. Deep sea water and Chelating agent  In the culture medium used By wavelength  Microalgae culture

1.1 Production of culture medium

It is important to manufacture low-cost, high-salt, high-nutrient enriched water that can grow microalgae such as dinaria in a culture technique that promotes useful substances such as beta-carotene. In addition, this process is disinfection and sterilization process by high temperature, and it is necessary to keep the concentrated water sent to the culture and transplanting apparatus clean, because it is a process to remove algae other than the algae to be cultured beforehand. There are very few algae and fungi surviving due to special environment such as deep sea water with no light and high pressure water. Approximately the fungal ratio of surface water and deep ocean water is about 9: 1 CPU / ml.

In addition, although very few, these bacteria and algae can be introduced into the seawater during the intake process and can grow in the culture apparatus. Some of the algae are beneficial, but some are harmful and some of the nutrients contained in the enrichment process may be consumed unnecessarily, thus removing some of them from the pretreatment process. The removal method removes suspended matters through a primary filtration process through a microfilter and a glass fiber filter. In natural seawater, iron, copper, and zinc, which are essential elements of photosynthetic organisms, exist, but they are also a limiting factor in basic production (Harold, 1999; Gerringa, 2000).

Ingredients of chelating agent Na ₂ EDTA Constituent density MaCl ₂ O 6 H ₂ O 1.5 g MaSO ₄ O 7H ₂ O 0.5 g KCl 0.2 g CaCl ₂ 쨌 2H ₂ O 0.2 g KNO ₃ One g NaHCO ₃ 0.043 g KH ₂ PO ₄ 0.035 g Fe-solution 10 ml Trace-element solution 10 ml Fe-solution (for 1L) Na ₂ EDTA 189 mg FeCl ₃ O 6H ₂ O 244 mg Trace-element solution (for 1L) H ₃ BO ₃ 61 mg (NH ₄₎ 6Mo ₇ O ₂₄ 4H ₂ O o 38 mg CuSO ₄ O 5H ₂ O 6 mg CoCl ₂ O 6 H ₂ O 5.1 mg ZnCl ₂ 4.1 mg MnCl ₂ O 4H ₂ O 4.1 mg

Therefore, it is necessary to stabilize the culture solution by adding a trace metal buffer such as EDTA. EDTA serves to remove from the medium where the high concentration of metal is not removed and regulates the supply of iron and the like. In this example, Na ₂ EDTA was added to the Johnsons medium to stabilize the metal elements and then used as a culture medium (Table 2). However, the chelating agent is not limited to Na ₂ EDTA, and the addition amount may vary depending on the concentration of concentrated water.

1.2 Cultivation of microalgae according to wavelength

 Dunaliella salina (KMCC 1064), a photosynthetic microalgae from Korea Marine Microalgae Bank, was inoculated 1/10 of the culture medium, and the ratio of lighting and lighting of the light source at f / 2 medium at 25 ℃ was measured L: D = 14: 10 and cultured in a 300 ml Erlenmeyer flask. Table 3 shows the conditions of the culture medium and the light source.

Condition of culture medium and light source wavelength Culture solution LED-blue (420-480 nm) Deep sea water + chelating agent (Na2EDTA) LED-orange (610nm) Deep sea water + chelating agent (Na2EDTA) LED-red (660nm) Deep sea water + chelating agent (Na2EDTA) Fluorescent lamp (2500lx) Deep sea water + chelating agent (Na2EDTA) LED-blue (450nm) Deep sea water + concentrated water (Johnsons medium) LED-orange (610nm) Deep sea water + concentrated water (Johnsons medium) LED-red (660nm) Deep sea water + concentrated water (Johnsons medium) Fluorescent lamp (2500lx) Deep sea water + concentrated water (Johnsons medium)

The light irradiation was performed using a light emitting diode, and the blue wavelength ranged from 420 to 480 nm with a maximum value of 450 nm, the red wavelength ranged from 640 to 680 nm with a 660 nm peak, and the yellow wavelength ranged from 580 to 620 nm with a 610 nm range , And the control was adjusted to 2500 lux by using a fluorescent lamp (FIG. 4).

1.3 Propagation changes of microalgae

After microalgae were propagated using light emitting diodes of different wavelengths, the proliferation change was measured by dry weight. Dry weight was measured by collecting 10 ml in a culture vessel once every two days, filtering with a GF / C filter, drying at 60 ° C for 24 hours, and heating to 0.001 g. All experiments were repeated 3 times.

As a result, the highest value was observed at 18 days in all cultures, but the increase was not significant after 8 days. Finally, at 0.04 g / 100 g on day 0 of culture, it was increased about three times on day 18 after culturing to 0.147 g / 100 g. There was a difference of about 0.04 g / 100 g in the experimental range of the blue wavelength and the red wavelength, but not in the other experimental period (FIG. 4).

1.4 Pigment changes in microalgae

The photosynthetic pigment of phytoplankton is chlorophyll a, b, c and auxiliary pigment. Of these, chlorophyll - a is included in all phytoplankton, so the biomass of phytoplankton can be best evaluated by measuring its amount. These pigments were extracted with a 90% acetone solution and the absorbance of the extract was measured at 750, 665, 645, 630 and 480 nm to calculate chlorophyll-a, total chlorophyll and plant carotenoid contents. The following is a calculation formula according to the Marine Environment Processing Test Standard (2010) for calculating chlorophyll-a, total chlorophyll and plant carotenoid content in the present invention. Here, E represents the absorbance value after the optimum absorbance correction and the turbidity correction, b represents the absorbance measurement value after the acid treatment, and 0 represents the absorbance measurement value before the acid treatment (FIG. 5).

Chlorophyll a = 11.6 E665 - 1.31 E645 - 0.14 E630

Chlorophyll b = 20.7 E645 - 4.34 E665 - 4.42 E630

Chlorophyll c = 55 E630 - 1.31 E665 - 0.14 E645

Total chlorophylls = Chlorophyll a + b + c

Plant Carotenoids = 4.0 E480

Phaeo-pigments = 26.7 (1.7 665b - 6650)

In addition to the phytoplankton living in seawater, photosynthetic pigments are also contained in the detritus, which are post-degraded, and the crust of animals predominating in phytoplankton. Therefore, in order to measure chlorophyll of living phytoplankton, it is necessary to measure phaeo-pigmant amount, which is an early decomposition product. In this experiment, the amount of phaeo-pigmant as well as chlorophyll-a and total chlorophyll were also analyzed, and all experiments were repeated three times.

In the case of chlorophyll-a, the content of the experimental period started to show from the mid-culture. At the blue wavelength, it was 1.02 ug / L, but at the red wavelength it was 0.27 ug / L. The difference was about 4 times. In the total chlorophyll, blue, yellow, and red wavelengths were observed in the mixed culture of deep sea water and chelating agent in the order of chlorophyll a and amount.

Therefore, in order to increase the chlorophyll content, it was found to be effective to cultivate blue wavelengths and fluorescent lamps. The phaeo-pigment content was 0.75 ug / L at the red wavelength, 1.86 ug / L at the yellow wavelength, 2.10 ug / L at the fluorescent lamp, and 2.90 ug / L at the blue wavelength, .

In the mixed culture of deep sea water and chelating agent, the content of carotenoids in plants was in the order of red, blue wavelength, fluorescent lamp and yellow wavelength. The changes in carotenoid content of plants were significantly increased from the start of cultivation. After 20 days of culture, blue, yellow, and fluorescent lamps showed 6.8-8.8 ug / L, while red wavelengths were 14.2 ug / L, indicating selective pigment production depending on wavelength (FIG. 6) .

2. Deep sea water and The culture medium  In the culture medium used By wavelength  Microalgae culture

2.1 Production of Culture Medium Using Seawater and Culture Medium

Some of the seawater algae are beneficial, but some of them are harmful and some of the nutrients contained in seawater (ocean deep seawater and deep seawater treated water) may be unnecessarily consumed, thus removing some of them in the pretreatment process. The removal method was the same as in Example 1.1, and the removed deep-sea water treated water was used as a base solution to prepare a culture medium containing a chelating agent. Therefore, only nutrients are added without adding sodium chloride, which is already used for seawater. In this experiment, Dunaliella Johnson ' s medium, which is used as a salina -specific medium, was added and used as a culture medium.

2.2 Proliferation of microalgae

The procedure of Example 1.1 was followed. The results are shown in Table 1. As in the case of Example 1, the results were highest at 18-20 days after culturing in all experimental groups, and did not show a significant increase after 8 days. Finally, after culturing at 0.06 g / 100 g on day 0 of culture, it was found to be about three times as high as 0.146 g / 100 g on day 18. In this experiment, there was no significant difference according to wavelength (FIG. 4).

2.3 Pigment changes in microalgae

And proceeded and measured in the same manner as in Example 1.4. The chlorophyll a content was 1.3 ug / L for blue wavelength, 1.7 ug / L for yellow wavelength, 0.6 ug / L for red wavelength and 1.2 ug / L for fluorescent lamp.

This result was slightly different from Example 1. Although the chlorophyll-a content is low and the content is high at other wavelengths at the red wavelength, the use of the culture medium is basically due to a large amount of nutrients required for growth, which is not limited to the wavelength 5).

This is because nitrate, phosphate, carbonate and various minerals are added to the culture medium. Of course, it is included in clean concentrated water basically, but the amount is increased, so it can be regarded as a difference depending on the nutritional composition. The total chlorophyll content showed a significant difference from the mid-culture to the experimental period. On the 20th day of culture, the blue wavelength was 2.6 ug / L, the yellow wavelength was 3.3 ug / L, the red wavelength was 1.2 ug / ug / L, which is similar to the chlorophyll a content.

Therefore, it is considered that the yellow wavelength is most suitable to increase the chlorophyll content, and the red wavelength can be selected to lower the chlorophyll content. In the case of phaeo-pigmant content, the experimental period started to show significant difference after 2 days of culture. In particular, on the 20th day of culture, no difference was observed between the blue wavelength of 3.8 ug / L and the fluorescent lamp of 3.7 ug / L, but the yellow wavelength of 4.7 ug / L and the red wavelength of 1.7 ug / there was.

The concentrations of carotenoids in plants were in the order of red, blue, yellow, and fluorescent lamps in the mixed cultures of deep sea water and culture medium. As with previous Example 1.1, the chlorophyll content was inversely proportional to the chlorophyll content. The carotenoid content tended to be higher at the red wavelength from the early stage of culture. Finally, the blue wavelength, the yellow wavelength and the fluorescence lamp showed 4.2 ~ 5.6 ug / L, while the red wavelength was 11.0 ug / (Fig. 6).

The amounts of chlorophyll and phaeo-pigmant, which are early decomposition products, were higher than those of mixed culture of deep seawater concentrated concentrate and chelating agent when the concentration of deep seawater and culture medium were higher than those of mixed culture of deep seawater and chelating agent. In case of plant carotenoid pigment, Water and chelating agent were higher than those of concentrated water and culture medium.

In order to increase the amount of chlorophyll and phaeo-pigmant, which is an initial degradation product, it is preferable to investigate the yellow wavelength using deep sea water and culture medium. In case of plant carotenoid pigment, deep seawater concentrate and chelating agent are used And it is judged that a good result can be obtained by examining the red wavelength.

3. Dunaliella saline Of pigment in salinity

The salinity was measured in f / 2 medium based on deep sea water (DSW), surface water (SSW), and surface water. The salinity was measured by the following method: NaCl 3.4%, NaCl 7.0%, NaCl 10%, NaCl 15%, NaCl 20% (CW + DSW), salt (10%), concentrated water and deep ocean water mixture (CW + DSW), salt 15%, concentrated water and deep sea water mixture (CW + DSW) (CW + DSW) and salt (25%), respectively. For each salt condition, 1/15 of the culture was inoculated and cultured for 24 hours in a glass tube to prevent cell aggregation. The stock culture was carried out at 25 ° C under fluorescent lamp conditions with L: D = 14: 10 in the f / 2 medium and the ratio of darkness and darkness to the total carotenoids. Chlorophyll a, total chlorophyll, phaeo-pigment, Respectively.

FIG. 7 shows the results of Dunaliella The change of chlorophyll pigment by salinity of salina is shown. In the case of chlorophyll-a, all of them showed a tendency to decrease on the 4th to 6th day, but they increased again on the 8th day and increased until 14th day, but the increase was less.

NaCl 3.4%> DSW ≥ SSW 20, 25%, it is thought that this is due to the effect of osmotic pressure since it is not detected from the 4th day after the inoculation. 10%> 15%> dense water, and 7% = 10%> 3.4%> 15% in the experimental group using the deep seawater concentrated water. 3.4%, and increased after 14 days. Except for f / 2 medium, 10% of concentrated water and 7% of NaCl showed similar tendency.

Fig. 8 is a graph showing the distribution of Dunaliella The change of total carotenoid pigment by salinity of salina is shown. In the case of total carotenoids, deep sea water (DSW) and surface water (SSW) did not show any difference and showed lower contents than 3.4% NaCl. The concentrations of chlorophyll - a and total carotenoids were 10%> concentrated water (7%)> 15% in the experimental water using concentrated water (CW). Especially on the fourth day, from 10% to 0.51 ug / L.

In the experiment using NaCl, 3.4%, 7% and 10% did not show difference on the 6th day, but more than 0.45ug / L in 7-10% on 8-10th day. Therefore, the high concentration of short - term deep seawater concentrate + salt was 10%.

Chlorophyll a content and total carotenoid content were not significantly different in the experimental group using sodium chloride, but 7 ≥ 10%> 15%. And 1.5 times higher than that of sodium chloride.

Chlorophyll a content and total carotenoid contents were higher in the order of 3.4% NaCl> DSW alone ≥ SSW alone in the order of salinity of 3.4% and 2 days and 12 days of culture.

By cultivating microalgae with deep seawater treated water (deep sea water concentrated in the ocean), which is a byproduct of the production of deep seawater, it is possible to use LED light to enhance useful components such as beta carotene Beta-carotene can be selectively and effectively obtained.

Claims (10)

(a) preparing seawater for microalgae culture;
(b) filtering, sterilizing and disinfecting seawater,
(c) culturing a microalgae resistant to salt in an incubator capable of regulating the culture environment;
(d) LED light with a wavelength of 420 to 480 nm is used to enhance phycocyanin while changing the culture environment and incubation period, LED light having a wavelength of 640 to 680 nm is used for beta-carotene enhancement, chlorophyll augmentation Characterized in that an LED light having a wavelength of 580 to 620 nm or a wavelength of 420 to 480 nm is used for the microalgae culture.
(a) preparing a deep seawater concentrated water for microalgae culture;
(b) adding a chelating agent to stabilize trace metals such as iron, copper, and zinc contained in seawater;
(c) filtering, sterilizing and disinfecting seawater,
(d) culturing at least one microalgae selected from the group consisting of dunaliella, chlorella, and spirulina in an incubator capable of controlling the culture environment;
(e) Culturing environment Microalgae culturing method for enhancing specific useful components, characterized by increasing the plant carotenoid pigment by culturing the cultured period for 2 to 14 days by irradiating a red wavelength at a salt concentration of 7-20% .
(a) preparing a deep seawater concentrated water for microalgae culture;
(b) adding a culture medium containing a buffer to stabilize trace metals such as iron, copper, and zinc contained in seawater;
(c) filtering, sterilizing and disinfecting seawater,
(d) culturing at least one microalgae selected from the group consisting of dunaliella, chlorella, and spirulina in an incubator capable of controlling the culture environment;
(e) Culture environment The specific effective component (s) is characterized by increasing the amount of chlorophyll and phaeo-pigmant, which is an initial degradation product, by culturing for 2 to 14 days after culturing the cultured period by examining the yellow wavelength with a salt concentration of 7-15% Of the microalgae. delete delete delete delete delete delete delete

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