KR100590296B1 - Method of Cultivating Chlorella - Google Patents

Abstract

In the present invention, in culturing chlorella using a culture medium containing nutrients such as nitrogen or phosphorus as a carbon source for the growth of chlorella as a carbon source to assist the photosynthesis of chlorella,

Iii) receiving 50% by weight of groundwater in a chlorella culture water tank and storing it in a natural state at 20 to 30 ° C. for a certain period of time (ground water aging step);

Ii) separately adding glucose and nutrients as carbon sources to 50% by weight of groundwater and storing the mixture for a period of time at 20 to 30 ° C. to prepare a culture solution (culture medium production step);

Iii) adding the culture solution prepared in step ii) to the aged ground water prepared in step iii) and culturing and propagating chlorella by photosynthetic action by natural light while stirring by impeller at 15-25 ° C. for a certain period of time (chlorella culture propagation) step);

It relates to a chlorella culture method characterized in that the culture through.

 In the present invention, the chlorella can be cultured without inoculating the chlorella fungus into the groundwater withdrawn, so it is not necessary to have a separate facility for aseptic work, and nutrients contained in the culture medium during chlorella culture do not precipitate in the culture solution. In order to dissolve evenly in the culture medium and to promote the photosynthesis action of chlorella to sufficiently absorb the dissolved nutrient oil, the culture solution is agitated by an impeller or the aeration of the culture medium is aerated to supply an air to the culture medium to chlorella culture. In order to accelerate the speed, conventionally, there is an effect of reducing the incubation period of 5 to 10 days to 3 days.

Chlorella, groundwater, glucose, nutrients, urea, magnesium sulfate, ammonium carbonate, potassium monophosphate, polyethylene, polypropylene, acrylic

Description

Chlorella cultivation method {Method of Cultivating Chlorella}

The present invention relates to a method for culturing chlorella, and more particularly, in a chlorella culturing method using a culture medium containing nutrients such as glucose and nitrogen or phosphorus as a carbon source, the growth rate of chlorella using natural light in groundwater taken underground It relates to a chlorella culture method characterized in that it can promote.

In general, chlorella is spherical or elliptical and rapidly proliferates 4 to 8 times every 20 hours through nuclear fission of its own cells, and has a very strong viability. Chlorella is a green algae plankton that dwells in fresh water such as lakes and contains large amounts of protein, dietary fiber, vitamins and minerals. Chlorella is 10 times more chlorophyll than normal vegetables and 10 times more photosynthetic. Alkaline foods are balanced with acidic constitution by overingestion of meat and grains with alkaline foods. It is also known to reduce action. In addition, calcium, zinc, and protein contained in chlorella function to prevent cadmium from being absorbed into the blood from the small intestine and to neutralize the toxicity of cadmium by producing low-molecular metal-bound proteins in the liver and kidneys, which are organs in which cadmium accumulates. Since it is known to make a variety of health supplements are being developed and marketed.

Therefore, many studies on chlorella cultivation methods have been conducted, and all of the above techniques use glucose as a carbon source for photosynthesis and nutrients such as nitrogen or phosphorus as a nutrient source for chlorella growth. Many patents have been filed for methods of inoculating and culturing chlorella in a sterile state.

In culturing chlorella in a medium containing saccharides and nitrogen in Korean Patent Publication No. 82-1310 (announced on July 21, 1982), the culture medium was added with 1.5 to 2.5% by weight of saccharifying enzyme and sterilized. In order to proliferate and cultivate chlorella by passing through O ₃ and irradiating with sunlight, it is known that the patent adopts a sterile culture method, so that a separate facility for supplying ozone to the culture solution is required, as well as culturing chlorella. There is a problem that the culture efficiency is low because the period takes 7 to 8 days, and the Republic of Korea Patent Publication No. 193748 (June 15, 1999) and Korea Patent Publication No. 19217 (March 15, 2001) In the production of chlorella by heterotrophic culture, glucose, such as crystalline glucose and glucose solution, is used as the main carbon source, and ammonia, urea, Incubate the culture by optimizing the initial sugar concentration within the range of 4 to 8% (W / V) and the inoculation amount within the range of 4 to 10% (v / v) in a sterile culture tank shaded using water as the main nitrogen source. In addition to artificial aeration, agitation, temperature and pH adjustment, at the time when the residue becomes 1 to 2% (W / V), additional sugar is added so that the residue becomes 3 to 4% (W / V). It is known that the cultivation method characterized in that the production, but the patent is because the chlorella is cultivated in a light-shielded sterile culture tank, as well as a separate installation for autoclaving at 121 ℃ high temperature for aseptic work There was a problem that the culture period of chlorella for use as a long time of 7 to 8 days and the culture efficiency such as proliferation for 5 days also very low.

Therefore, in order to solve the above problems, the present invention provides a carbon source for assisting chlorella's photosynthesis, and in culturing chlorella using a culture medium containing nutrients such as nitrogen or phosphorus as a nutrient source for growth of chlorella and chlorella. In addition, since the chlorella can be cultured without inoculation of the chlorella fungus, it is an object of the present invention to provide a chlorella culturing method characterized in that it does not need to have a separate facility for aseptic work.

In the present invention, in the culturing of chlorella using a culture medium containing nutrients such as phosphorus and nitrogen as a carbon source for the growth of chlorella as a carbon source to assist the photosynthesis action of chlorella, the nutrients in the culture medium In order to dissolve evenly in the culture medium without being precipitated, and to promote the photosynthesis action of chlorella to sufficiently absorb the dissolved nutrient oil, the culture medium is agitated by an impeller, or the culture medium is aerated in an acid pipe supplying air into the culture medium. Another object of the present invention is to provide a method for culturing chlorella, characterized in that it can promote the culturing rate of chlorella.

In the present invention, in culturing chlorella using groundwater of natural intake without sterile operation, the incubation period of 5 to 10 days can be shortened to 3 days. There is a characteristic.

The present invention relates to a method for culturing chlorella, and more particularly, to cultivate chlorella using a culture medium containing nutrients such as nitrogen or phosphorus as a carbon source to assist chlorella's photosynthesis, and a nutrient source for chlorella growth. The present invention relates to a chlorella culturing method characterized by rapidly culturing chlorella without inoculation of chlorella fungi by using natural light in groundwater taken underground.

In the present invention, unlike the chlorella culturing method of inoculating chlorella fungi in a sterile state and then basing the culture, the present invention does not need to sterilize the groundwater taken from the basement without leaving the sterilization treatment at room temperature for 3 to 6 days before the growth of chlorella. If nutrients such as nitrogen or phosphorus are added to glucose and nutrient sources, which are necessary carbon sources, and then aerated at 25 to 35 ° C. for rapid cultivation, chlorella is excellent in photosynthetic capacity and 4 to 20 times every 20 hours through nuclear fission of its own cells. Because it multiplies very fast, 8 times, and has very strong vitality, after a certain period of time, the growth of other fungi is suppressed, and only chlorella grows.

Hereinafter, the chlorella culture method of the present invention will be described in detail.

In culturing chlorella using a medium containing glucose as a carbon source to assist chlorella's photosynthesis and a nutrient such as nitrogen or phosphorus as a nutrient source for chlorella growth,

Iii) receiving 50% by weight of groundwater in a chlorella culture water tank and storing it in a natural state at 20 to 30 ° C. for a certain period of time (ground water aging step);

Ii) separately adding glucose and nutrients as carbon sources to 50% by weight of groundwater and storing the mixture for a period of time at 20 to 30 ° C. to prepare a culture solution (culture medium production step);

Iii) adding the culture solution prepared in step ii) to the aged ground water prepared in step iii) and culturing and propagating chlorella by photosynthetic action by natural light while stirring by impeller at 15-25 ° C. for a certain period of time (chlorella culture propagation) step);

Chlorella is cultured through.

In step iii), the groundwater is stored in a chlorella culture tank at room temperature for a certain period of time to mature groundwater. The chlorella culture tank used at this time is made of a transparent material in front and rear and left and right side walls so that sunlight can pass through the culture tank. , Glass or polyethylene resin, polypropylene resin, acrylic resin may be used. And the maturation period of the groundwater is yellowed in 2 to 3 days in summer and 5 to 6 days in winter. The color of the groundwater changes yellow because various fungi contained in the groundwater multiply. At this time, the chlorella fungus multiplies with other fungi.

Step ii) is a step of preparing a culture solution by adding glucose and various nutrients to the groundwater, wherein the amount of nutrients added to the groundwater is as follows.

The amount of glucose, which is a carbon source, is added to 0.2 to 0.4% by weight of the total amount of groundwater, and the amount of nutrients is 0.3 to 0.6% by weight of the amount of urea, 0.2 to 0.4% by weight of magnesium sulfate, Ammonium carbonate is dissolved by adding 0.2 to 0.4% by weight of the total amount of ground water and potassium monophosphate is added by 0.2 to 0.4% by weight of the total amount of ground water.

In the above glucose, chlorella, especially on a cloudy day or when it is rainy, because the photosynthetic action of chlorella is not active and can not properly receive the supply of carbon at this time serves as a carbon source necessary for the growth of chlorella. If the added amount is less than 0.2% by weight, the amount of carbon necessary for the growth of chlorella is not sufficiently supplied, and if it exceeds 0.4% by weight, it does not help much for the growth of chlorella.

In the above, urea serves as a source of nitrogen components necessary for nitrogen chlorination of chlorella, and when the amount is less than 0.3 wt%, the amount of nitrogen necessary for the growth of chlorella is not sufficiently supplied, and the growth of chlorella is greater than 0.6 wt%. It doesn't help much.

Magnesium sulfate is a basic constituent of chlorophyll of plants, which promotes photosynthesis to increase starch quality, thus promoting chlorophyll formation, increasing yield, and facilitating phosphate transport at the growth point where phosphoric acid is absorbed and cell division is active. When the addition amount is less than 0.2% by weight of chlorophyll is not increased because the growth of chlorella is difficult, and even more than 0.4% by weight does not help the growth of chlorella is not economical.

Ammonium carbonate serves to dissociate in the culture medium to produce ammonium ions, which are carbon dioxide and nitrogen sources necessary for photosynthesis. If the added amount is less than 0.2% by weight, the amount of carbon dioxide generated in the culture medium is insufficient, so that photosynthetic action cannot be progressed smoothly, and the proliferation of chlorella is suppressed. Even if it does occur, it does not help the growth of chlorella.

The potassium monophosphate is a nutrient necessary for the growth of chlorella, and serves to increase the protein content in the chlorella. If the amount is less than 0.2% by weight, the protein content in the chlorella decreases due to the lack of nutrients necessary for the growth of the chlorella. Even if it exceeds 0.4% by weight, the protein content in the chlorella does not increase particularly.

The culture solution prepared in step ii) is stored for 1 day to sufficiently dissolve glucose and various nutrients added to the culture solution so that chlorella can absorb it well.

The optimum temperature of the groundwater of step iii) is 20-30 ° C, which is the optimum temperature of the initial fungus activity, and ii) the optimum temperature is 20-30 ° C because the culture medium must also be mixed with the groundwater of step iii). to be.

In the step iii), the culture rate of chlorella is increased by agitating the culture solution by an impeller or by aspirating the culture medium to an air supplying air in the culture medium in order to promote the photosynthesis action of the chlorella to sufficiently absorb the dissolved nutrient oil. Will be promoted. When the culture solution turns green after 2 to 3 days at 15 to 25 ° C., the culture growth of chlorella is completed. The population of the grown chlorella is 1,000 to 15 million / 500 L.

The optimum temperature at which chlorella is most active for photosynthesis is 15-25 ° C., where chlorella undergoes rapid proliferation culture. When chlorella is cultured and grown at room temperature, the chlorella continues to grow and perform vigorously photosynthesis. Therefore, the chlorella is killed before harvesting chlorella due to the lack of carbon dioxide and various nutrients in the culture medium. When stored at a low temperature of ℃ chlorella activity is inhibited as much as possible no longer proliferate, so the chlorella will not die until harvest.

(Examples 1 to 3)

1) After receiving 500ℓ of groundwater taken from 200m underground and storing it in 1,000l culture tank formed with glass walls on the left and right sides, and storing it at room temperature for 3 days, fungi contained in the groundwater multiply by photosynthetic action by natural light. The color changes to yellow. At this time, the groundwater is not subjected to autoclaving.

2) Separately, take 500 liters of groundwater and add glucose as a carbon source, urea, magnesium sulfate, ammonium carbonate, and potassium monophosphate as carbon sources, respectively, as shown in Examples 1 to 3 below. It is dissolved and aged for one day at room temperature so that Chlorella can absorb the nutrients well to prepare a culture solution.

3) The culture solution prepared in step 2) is mixed with the culture water tank of step 1) and added to the aged ground water, and then stored for three days with sufficient mixing with an impeller. The culture solution turns green, and cultivation of chlorella is completed. . The cultured chlorella is stored at a low temperature of 5 ° C. until shipment to inhibit chlorella's photosynthetic activity.

(Comparative Examples 1 to 3)

By adopting the same method as in Examples 1 to 3, the chlorella is cultured, but the groundwater taken in step 1) and the culture solution prepared in step 2) are sterilized at 120 ° C. for 20 minutes, and the growth of step 3) is performed. It was carried out in the dark. Proliferation cultured chlorella was stored at room temperature. At this time, the amount of the nutrients added to the culture solution was added in the same amount as in Examples 1 to 3, and is as described in Table 1 below.

TABLE 1

                                                            (Unit: kg / 500ℓ)

division Example Comparative example One 2 3 One 2 3 glucose 2 3 4 2 3 4 Element 3 5 6 3 5 6 Magnesium sulfate 2 3 4 2 3 4 Ammonium Carbonate 2 3 4 2 3 4 Potassium monophosphate 2 3 4 2 3 4

                                 TABLE 2

(Unit: 10,000)

division Example Comparative example One 2 3 One 2 3 Chlorella Individuals (Number of Individuals / 500ℓ) 1,026 1,248 1,497 315 369 402

In Table 2, the amount of the protein in the culture medium was measured to show the results of calculating the population by the size of the chlorella (based on 2㎛).

As a result of [Table 1], as a result of culturing and growing in a culture medium to which various nutrients were added, in the case of Examples 1 to 3, chlorella was cultured in a natural state without sterilizing the groundwater taken from the added amount of nutrients. It can be seen that the proportion of chlorella grows in proportion, but in the case of Comparative Examples 1 to 3, chlorella population compared to Examples 1 to 3 as a result of culturing in cows after autoclaving to keep the groundwater and the culture solution aseptically maintained. You can see that much less. However, even in comparison, it was found that the chlorella population also increased in proportion to the amount of nutrients added as in the example.

In addition, in the case of Examples 1 to 3 after the growth of chlorella was completed, chlorella survived after 5 days as a result of inhibiting chlorella's photosynthetic activity at a low temperature of 5 ° C., but in Comparative Examples 1 to 3, After 5 days at room temperature after the completion of culture propagation, it was confirmed that chlorella died.

In the present invention, in culturing chlorella, chlorella can be cultivated without inoculating chlorella fungus into the groundwater withdrawn, and therefore, there is no need for a separate facility for aseptic work, and nutrients contained in the culture medium during chlorella culture. In order to promote the photosynthesis action of chlorella and to sufficiently absorb the dissolved nutrient oils without being precipitated in this culture and to promote the photosynthesis action of the chlorella, the culture solution is stirred by an impeller or the culture medium is supplied to an air incubator that supplies air into the culture solution Since aeration to promote the culture rate of chlorella has the effect that can be shortened to three days in the conventional culture period of 5 to 10 days.

Claims (8)

In culturing chlorella using a medium containing glucose as a carbon source to assist chlorella's photosynthesis and a nutrient such as nitrogen or phosphorus as a nutrient source for chlorella growth, Iii) receiving 50% by weight of groundwater in a chlorella culture water tank and storing it in a natural state at 20 to 30 ° C. for a certain period of time (ground water aging step); Ii) separately adding glucose and nutrients as carbon sources to 50% by weight of groundwater and storing the mixture for a period of time at 20 to 30 ° C. to prepare a culture solution (culture medium production step); Iii) adding the culture solution prepared in step ii) to the aged ground water prepared in step iii) and culturing and propagating chlorella by photosynthetic action by natural light while stirring by impeller at 15-25 ° C. for a certain period of time (chlorella culture propagation) step); Chlorella culture method, characterized in that the culture. The method of claim 1, wherein the chlorella culture tank used in the step iii) is a transparent material in front, rear, and left and right sidewalls so that sunlight can pass through the culture tank, and among the glass or polyethylene resin, polypropylene resin, and acrylic resin. Chlorella culture method, characterized in that using one selected. According to claim 1, Chlorella culture method characterized in that the storage period of the groundwater in the step iii) is 2 to 3 days in the summer. According to claim 1, Chlorella culture method characterized in that the storage period of the groundwater in the step iii) 5 to 6 days in winter. The method of claim 1, wherein the addition amount of glucose as a carbon source in step ii) is 0.2 to 0.4% by weight of the total ground water usage. According to claim 1, wherein the amount of nutrients added in step ii) is 0.3 to 0.6% by weight of the total ground water consumption of urea, 0.2 to 0.4% by weight of the total ground water usage of magnesium sulfate, 0.2 to 0.2% of the total ground water usage of ammonium carbonate Chlorella culture method, characterized in that 0.4% by weight, potassium monophosphate is 0.2 to 0.4% by weight of the total ground water usage. The method of claim 1, wherein in step ii), the culture solution is stored for one day, and the glucose and various nutrients added to the culture solution are sufficiently dissolved so that chlorella can absorb well. The method of claim 1, wherein the culture in step iii) Chlorella culture method, characterized in that stored for 2 to 3 days.