KR100876614B1 - Flotation induction and harvesting of Arthrospira platensis KCTC 11039 - Google Patents

Abstract

The present invention relates to the induction and harvesting method of arthrospa pulmonary injury, and more particularly to the cyanobacteria Arthrospira which is used as a dietary supplement, bioactive substances, livestock and pet food. A method for inducing the injury of Arstrophira spp. By adding a certain amount of sodium carbonate to the existing culture water which makes it difficult to economically harvest as the flocculation capacity of Arsphrodera decreases depending on the culture conditions. will be.

Arthrospira formation, injury induction, harvesting, sodium carbonate

Description

Flotation induction and harvesting of Arthrospira platensis KCTC 11039}

Figure 1 shows a micrograph of Arthrospira platensis KCTC 11039BP.

Figure 2a shows the addition of 0.025, 0.05, 0.1, 0.2, 0.3 M test spheres from the left side of sodium carbonate to the Arthrospyra plethesis KCTC 11039BP culture sample by concentration, and after 12 hours, Arthrospyra plethesis KCTC 11039BP is a photograph showing the degree of injury of the body,

FIG. 2B shows the incidence of Arthrospyra condyle after 12 hours of addition of 0.025, 0.05, 0.1, 0.2, and 0.3 M test zones from the left side of sodium bicarbonate to the Arsspira ple tensis KCTC 11039BP culture sample. Shows a photograph showing the degree,

Figure 2c shows the addition of 0.025, 0.05, 0.1, 0.2, 0.3 M test zone, control (far left) of sodium chloride in the Arthrospyra plethesis KCTC 11039BP culture sample by concentration and after 12 hours This is a photograph showing the degree of injury of Arstrophira.

3 is a graph showing the degree of injury of Figures 2a, 2b, 2c as a numerical value (injury rate,%).

Figure 4 is a graph showing the rate of injury (%) over time after the addition of sodium carbonate to the Arthrospyira plethesis KCTC 11039BP samples by concentration.

Figure 5 is a photograph showing the appearance (a) and injured appearance (b) of the in-situ mass culture of Arthrospyira plestinis KCTC 11039BP.

Fig. 6 is a schematic diagram showing a harvesting device of the floating Arthrospyra plethesis KCTC 11039BP aggregates.

[Description of Symbols for Main Parts of Drawing]

 1: channel type outdoor incubator 2: blower

 3: switchgear 4: filter network

The present invention relates to the induction and harvesting method of arthrospa pulmonary injury, and more particularly to the cyanobacteria Arthrospira which is used as a dietary supplement, bioactive substances, livestock and pet food. A method for inducing the injury of Arstrophira spp. By adding a certain amount of sodium carbonate to the existing culture water which makes it difficult to economically harvest as the flocculation capacity of Arsphrodera decreases depending on the culture conditions. will be.

Arthrospira is a coil wound in the form of a long filament as shown in Figure 1, the cell width is 3 ~ 16㎛, the length between the coil is 10 ~ 70 ㎛, the diameter of the coil 20 to 100 μm, cells and cells are connected in septum (septum) unit.

Arsroth containing abundant protein content (60 ~ 70%), vitamins (vitamin B12, provitamin A), essential amino acids, minerals and essential fatty acids, additives and dietary supplements for pet food, aquaculture feed Microalgae are the world's highest-produced microalgae [Belay, A., 2004, The potential application of Spirulina ( Arthrospira ) as a nutritional and therapeutic supplement in health management]. According to recent estimates, Arthrospy is the world's largest producer of microalgae, producing approximately 4000 tons per year. A major problem in the industrial production of microalgae, including arthrospyira, is the productivity and economics of harvesting. The economic cost of harvesting exceeds 20% of the total cost.

In the case of arthrospyra, it is known to perform vertical migration including air vacuole in cells. In other words, during the day it repeats the pattern to rise to the water surface for photosynthesis and to settle at night.

Harvesting from mass cultivation of Arthrospyra is mainly by filtration, but it causes a lot of difference in filtration efficiency depending on cell length and requires a lot of time and effort. Harvesting by the treatment of the flocculant requires a lot of processes such as the treatment of the supernatant and removal of the flocculant component, and harvesting by the centrifugal is difficult to completely harvest due to the flotation of arthrospira and incurs many economic costs. In addition, the coagulation, filtration, and harvesting by centrifugation requires the preparation of ancillary facilities because the whole culture must be processed. The harvest of Arthrospyra in the outdoor cultivation site is left overnight, with all lights, aberrations, and air pumps turned off to take advantage of Arthrospyra's flotation, and naturally rises the next morning to form a thick layer. A simple method of harvesting an arthrospyra is also used. Therefore, using the natural flotation of Arthrospyra to harvest the flocculated chunks on the surface of the culture water with a net-like instrument would be the most economical and the method with little change in the physical properties of the Arthrospyras. . Korean Laid-Open Patent No. 2005-0030340 is a patent using the flotation of Arsspira (or Spirulina, also called Spirulina ), and it is a method that can be easily harvested by adding sodium chloride to the culture solution of Arsrospyra to improve the flotation activity. Presented. In other words, by improving the flocculation of the tank in the arthrospy culture tank, it is possible to easily recover the bath suspended in the upper layer of the culture tank. However, the problem of the Republic of Korea Patent Publication No. 2005-0030340 is that the precipitate can be formed in the culture by adding a large amount of sodium chloride of 1 to 2%, has a problem that it is difficult to reuse the culture by high salt. Harvesting after culturing arthrospyra for a short period of time is economically possible with the addition of salt because the physiological conditions of arsspira are relatively good. You will not be able to induce enough injury to harvest. Inflammatory deterioration of arthrospira is a common phenomenon in long term culture. In order to improve the light utilization of the cultivated arthrosphora water is excessively circulated by aberration or lack of nutrients, the flotation of arthrospy is very low, and the harvesting takes much time and expense, but the harvesting efficiency is low.

Accordingly, the present inventors have conducted research in order to solve the above problems, and as a result, by adding sodium carbonate to a specific concentration of arthroscopy culture water and culturing the long-term culture of arthroscopy using a water-outdoor incubator equipped with a blower In this case, even when the flocculation of arthrospy is significantly reduced, the present invention has been completed by developing a method of inducing the flossing of the arthrospy granules in a short period of time and making the harvesting simple and economical.

Accordingly, an object of the present invention is to provide a method for inducing the flotation of arthrospy granules in a short period of time even in an outdoor mass incubator and enabling a simple and economical harvesting.

The present invention is a method for inducing the injury of the Arthrospyras by adding 0.3-0.4M of sodium carbonate to the culture water of Arthrospira platensis KCTC 11039BP and swelling under dark conditions for 6-24 hours. It is characterized by.

In addition, the present invention, by using the blower (2) installed on one side of the waterway type outdoor incubator (1), the arthrospy aggregates floating on the surface of the culture water is concentrated to the harvesting place and the other side of the incubator (blower) Opposite) is characterized in that the method of harvesting the Arthrospy aggregates with a 20 ~ 30 ㎛ mesh network 4 of the lower end of the opening and closing device (3).

The present invention will be described in more detail as follows.

The present invention is economical harvesting as the floating ability of the Arthrospyras aggregates lowered according to the culture conditions of the cyanobacteria Arthrospyra ( Arthrospira ), which is used as a dietary supplement, bioactive substances, livestock and pet foods The present invention relates to a method for inducing the injury of Arthrospyra by adding a certain amount of sodium carbonate to the existing culture water which makes it difficult to improve the ability of Arthrospyra in a short time.

Harvesting, which is one of the most important factors affecting the economics of industrial mass production of arthrospyra, or long-term cultivation, has greatly reduced the floating activity of arthrospy, making effective harvesting a very difficult problem.

In order to solve this problem, the present inventors have established a reagent and an optimized concentration range which can shorten the induction time of the injury activity of Arthrospira . Arthrospyra strains (Arrosspira pletensis KCTC 11039BP) were distributed from Korea Research Institute of Bioscience and Biotechnology, and the composition of SOT medium except CaCl ₂ ㆍ 2H ₂ O in groundwater using an outdoor waterway mass culture tank (5.7 ton) (Table 1 ) Was incubated for 30 to 50 days. After incubation, 0.3 to 0.4M of Na ₂ CO ₃ was added to a portion of the culture water, and the mixture was allowed to stand for 6 to 24 hours under dark conditions.

As described above, it was confirmed that when physiological incubation was performed by adding sodium carbonate in a specific ratio to Arsspira plethesis KCTC 11039BP culture water, induction of over 90% of the body injury within 6 to 24 hours.

 Therefore, the present invention is characterized by a method of inducing the injury of Arthrospyras by adding 0.3-0.4M of sodium carbonate to the culture water of Arthrospyira plethesis KCTC 11039BP, and then standing under the dark conditions for 6 to 24 hours. do.

In addition, the present invention by installing a blower on one side of the waterway-type outdoor incubator for algae culture to collect the injured arthrospy aggregate in one direction, collected by the opening and closing device installed on the other side of the incubator (opposite the blower) It is possible to harvest a large amount of Arstrophira by simply using a 20-30 μm mesh filter (bottom opening and closing device) so that the tank can be filtered by passing through the prepared tank without changing the physical properties of the tank.

That is, according to the present invention, by using the blower (2) installed on one side of the waterway type outdoor incubator (1), the arthrospy aggregates floating on the surface of the culture water are concentrated to the harvesting place, and the other side of the incubator (the blower) Opposite) is characterized in that the method of harvesting the spirulina with the sieve 4 of the 20 ~ 30 ㎛ mesh wood at the bottom of the switchgear (3) is another feature.

As described above, the present invention can shorten the conventional Arthrosphyra culture time and can be easily harvested. In addition, it is expected that not only arsspira but also various floating algae may be applied.

Hereinafter, the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited to these Examples.

Example  1: long-term culture Arthrospyra Constitutional Flotation  Judo

In order to induce injury in Arthrospira plethesis KCTC 11039BP cells with markedly decreased flotation activity, salts (NaCl), NaHCO ₃ and Na ₂ CO ₃ , which are known to enhance the flocculation of Arthrospyras, have been used. Used. Arthrospira samples were injected into the corning tube by 10 ml and each reagent was 0.025, 0.05, 0.1. After addition at the concentration of 0.2 and 0.3 M, it was dark-treated in the state left still at room temperature for 12 hours. After 12 hours of incubation with each reagent, Arsspira showed marked flotation in the sections treated with Na ₂ CO ₃ 0.2 and 0.3 M as shown in FIG. 2A and in the remaining reagent treatment groups (FIGS. 2B and 2C) It was found to have settled to the bottom as in the control (FIG. 2C leftmost tube). This result was measured using the following equation (1) flotation.

1 ml of sample was sampled 2 cm high from the bottom of the Corning tube and was measured for absorbance by measuring absorbance at 680 nm. The results are shown in FIG. Na ₂ CO ₃ injuries 96.9% in the 0.3M section, leading to the highest injury and 0.22.5% injuries. At concentrations below 0.2 M, some rose to the top, but settled to the bottom. In NaHCO ₃ section, some injury occurred in 0.3 M section, but it settled more than 22% compared to the control group, and the rest section settled overall. In NaCl addition section, there was some injury in 0.3 M section, but it settled 16.3% more than the control group, and most of the remaining sections showed the result. Therefore, the addition of Na ₂ CO ₃ induced markedly increased injury in injured Arthrospyra cells, which significantly slowed down the incidence, especially in excess of 92.5% of Arthrospy aggregates above 0.2 M Na ₂ CO _3. . The natural injury of Arthrospyra is usually settled in large amounts at night and then injured temporarily by daylight. However, when 0.2 M and 0.3 M Na ₂ CO ₃ were added, more than about 93% of the injuries had already been injured before exposure to light. This may be due to the Na ₂ CO ₃ concentration added, not due to the floating activity of the arthrospira cells themselves. Na ₂ CO ₃ is commonly used as a reagent to increase the pH in the purpose of controlling contaminants in culturing arthroscopy. In order to investigate whether the pH had an effect on the flotation of the Arthrospyras, the pH of the experimental section was examined in Table 2 below. The Na ₂ CO ₃ treatment was higher than the other experiments with pH of 9.3 or higher, and the induction of 93% or higher flotation was 10.4 or higher. These results indicate that pH can influence the induction of injury of the Arthrospy morphology.

Example 2: Optimal Na for Induction of Injury of Arthrospy Conjugates ₂ CO ₃  Amount

The long-term culture which Ars optimal Na ₂ CO Spirra play Ten sheath KCTC 11039BP by adding the number of the culture over time efficiency of the specific portion to the Ars the Na ₂ CO ₃ at different concentrations to determine the _third amount for deriving a portion of Spira in Investigate. Arthrospira plethesis KCTC 11039BP culture water was filled with 40 ml of the same sample as in Example 1 in a 50-ml plastic tube, and added to the Na ₂ CO ₃ concentration of 0.2, 0.3, 0.4, 0.5M and allowed to stand at room temperature. We investigated the incidence of Arthrospy tense over time. The flocculation of the Arthrospyras was investigated in the same manner as in Example 1 by sampling 1 ml at 2 cm from the bottom of the plastic tube. The results of the experiment are shown in Figure 3. Injury of Arthrospyra was induced in all Na ₂ CO ₃ concentrations except control, and after 1 hour, more than 64% of injuries were injured in 0.2 M Na ₂ CO ₃ . Injuries of the arthrospy aggregates increased with increasing Na ₂ CO ₃ concentrations, peaking at 0.4 M and slowing at higher concentrations (0.5 M). As time passed, the floss of Arthrospy granules increased, reaching about 90% at a concentration of 0.4 M after 6 hours. The slowing of the injury of the Arthrospyras at 0.5 M was determined to be due to the addition of Na ₂ CO ₃ to the sample more than the appropriate amount, which significantly increased the pH, thereby affecting the physical properties of the cells. The pH of Na ₂ CO ₃ concentration range was 0.2.10.58, 0.3M was 10.75, 0.4M was 10.85, 0.5M was 10.93 and the control was 10.3. This pH was different from that in Example 1. That is, in Example 1, the best results were obtained at pH 10.4 and 10.5, while in Example 2, the pH of the Na ₂ CO ₃ concentration of 0.2M was high as about 10.6, but the induction of injury of the arthrospy complex was low. Thus, induction of the floss of Arthrospyra through Examples 1 and 2 is not simply caused by pH and increases of carbon dioxide and sodium (Na ⁺ ) ions and OH ⁻ ions generated by dissolution of Na ₂ CO ₃ . It is believed that the increase contributed to the induction of injury of the Arthrospy tense. Therefore, the results derived from Examples 1 and 2 indicate that the addition of Na ₂ CO ₃ to the Arthrospyura plethesis KCTC 11039BP culture water to a concentration of 0.3 to 0.4M is the most effective in inducing the injury of Arthrospyras. Less than 0.3M requires a lot of time to induce flotation, whereas concentrations above 0.4M can lead to a change in the properties of the Arthrospy trough, thus slowing induction of injury. 0.3 ml of Na ₂ CO ₃ may be appropriate when overnight culture of Arthrospyra plethesis KCTC 11039BP is required, and 0.4 M is appropriate when inducing the injury of Arthrospyra in a short time.

Example 3 Induction and Harvesting Arthrospy Stem Injury

Examples 1 and 2 in order to harvest the Spira to Ars that the culture in the field from the results of Ars, located in Gongju janggimyeon by the addition of Na ₂ CO ₃ 0.3M in Chapter Spira outdoor mass culture overnight after a political Spira to Ars crude Induced injury and harvested in an easy way. Arthrospyra was incubated for 50 days with SOT medium composition in groundwater. During the cultivation, the evaporated water was replenished and incubated with natural light. The culture water was circulated at a speed of 1.2 m / sec using aberration, and the aberration was continuously operated for 3 hours at 20 minute intervals. Air was continuously vented using an air pump. In order to harvest the arthrospy granules, the concentration of Na ₂ CO ₃ was added to the cultured water over 5 hours to 1 hour, and then allowed to stand until the next morning without running aberration and air pump. 5a shows the state before the incubation of the culture water and FIG. 5b shows the state after the stationary overnight. Before standing, it was confirmed that the arthrospy complex was well distributed throughout the cultured water, and after standing overnight, it formed a thick layer of arthrospy granules of 0.5 cm or more. At this time, the flotation efficiency of the Arthrosphyra body reached 98%.

In order to harvest the resulting rise of Arthrospyras was harvested by the incubator as shown in FIG. The harvested injured body can be blown in one direction of the outdoor incubator 1 by using a device capable of blowing a fan, such as a fan, so that the injured arthrospy body on the surface of the culture water can be sent in one direction. do. Install the opening and closing device (3) to the appropriate size above the end of the incubator, and placed under the 20 ㎛ mesh size filter net (4) it is possible to easily harvest the aggregated Arthrospy aggregates.

In reality, the harvesting of arthrospyra is most common, with a three-stage incidental filtration from the overall primary filtration of the culture [Belay, A., 2004, The potential application of Spirulina (Arthrospira) as a nutritional and therapeutic supplement in health management]. This process causes damage to the Arthrosphyra complex and decreases the filtration efficiency. On the other hand, the harvesting method of the present invention is a simple blower, only the body that emerged as a surface layer of cultured water gathered toward the collecting device, the first harvest is made, and the harvest is completed while washing with distilled water. Thus, induction of flotation by the present invention is a method that enables an effective and economical harvesting of the Arthrospy aggregates.

As described above, the efficiency of harvesting, which is one of the most important factors affecting the economics in the industrial mass production of arthrospyra, or the long-term culture, significantly lowers the floating activity of arthrospyra, so that effective harvesting is very difficult. Is emerging. In the present invention, by adding a concentration of 0.3 ~ 0.4M of Na ₂ CO ₃ to the culture water of arthrospyra, more than 90% of the arthrospy aggregates can be induced to rise to the surface of the culture water, and the incubator of FIG. By using the economical and simple physical properties of the unchanged can be harvested easily. In this way, the method of inducing injury by adding Na ₂ CO ₃ is expected to be applicable not only to arsspira but also to various floating algae.

Claims (3)

delete Using a blower (2) installed on one side of the waterway type outdoor incubator (1), the spirulina and the floating body on the surface of the culture water are concentrated at the harvesting place, and the switch is installed on the other side of the incubator (opposite the blower). (3) A harvesting method of arthrospy granules, characterized in that harvesting with a 20 ~ 30 ㎛ mesh network of the lower end (4). The method of claim 2, wherein the spirulina is characterized in that the injury induced by the addition of 0.3 ~ 0.4M sodium carbonate to the culture water of Arthrospira platensis KCTC 11039BP after standing for 6 to 24 hours under dark conditions Harvesting method.

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