KR20160085240A - Method of and Apparatus for Producing Biomass with Glycerol - Google Patents

Abstract

The present invention provides a method for culturing biomass using glycerol, and an apparatus for treating the same. According to the present invention, after specific microalgae are selected and cultured, a complex nutrient is inserted from the outside with respect to the cultured microalgae, thereby the microalgae are induced to photosynthesize to grow and absorb the complex nutrient to grow at the same time. The present invention has an effect of improving an output of microalgae by about 30 to 40 % when the complex nutrient is inserted, in comparison with the case in which the complex nutrient is not inserted.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a method and apparatus for growing biomass using glycerol,

The present invention relates to a method and apparatus for growing biomass using glycerol, and more particularly, to a method and apparatus for growing biomass using glycerol, and more particularly, The present invention relates to a method for growing biomass using glycerol and a proliferation apparatus therefor.

Today, efforts are being made to replace conventional petroleum products in various aspects in terms of concern about exhaustion of fossil fuels and prevention of environmental pollution.

What is perceived as the most viable fuel to replace conventional gasoline or diesel is biofuels produced through biomass, a renewable renewable resource. Bioethanol and biodiesel, already used in Brazil and other countries, are recognized as alternative fuels.

Biodiesel is an alternative to petroleum-based diesel oil and is based on renewable resources such as vegetable oils and animal fats. Biodiesel is composed of fatty acid methyl esters and contains two oxygen atoms in the ester functional group. Oxygenated fuel such as biodiesel is advantageous in that the oxygen contained in the molecule during combustion contributes to the combustion reaction, facilitating complete combustion and thus reducing the exhaust gas.

Biodiesel is considered a viable alternative to fossil fuels, which is the most important transportation energy resource, because it can replace current engine oil as a renewable fuel and can be transported and sold through existing facilities. However, biodiesel is still mixed with diesel to improve the low lubrication of pure ultra low sulfur fuel. The production and use of biodiesel is growing rapidly, especially in Europe, the US and Asia, but the share of the total fuel market is still insignificant.

Biodiesel can be produced using rapeseed, grains, farms, corn, etc. However, with the food problems on the planet now unresolved, production of food resources into biomass is not only an efficient use of resources, It is impossible to be free.

From this point of view, the production of biomass using microalgae has no problem of environmental pollution, and is independent of the acquisition of human food resources, so there is no ethical problem at all.

However, the productivity of microalgae is still only 1/5 - 1/10 of that of conventional fossil fuels. In recent years, research on the development of lipid-high productivity microalgae using biotechnology has been conducted along with exploration of excellent cell lines, but no remarkable results have been reported yet.

Korean Patent No. 10-1363723 "Method for Extracting and Recovering Lipids from Microalgae" (Feb. Korean Patent No. 10-1134294 "Oil Extraction from Microalgae and Biodiesel Conversion Method" (Apr. 2, 2012); Korean Patent No. 10-1264543 "Method for extracting raw material oil for biodiesel from microalgae and method for producing biodiesel using the same" (Feb.

Disclosure of Invention Technical Problem [8] The present invention has been made to solve the above problems of the prior art, and it is an object of the present invention to provide a biosurfactant for biomass which can grow a large amount of biomass as a means for securing alternative energy, The present invention provides a method of propagating a biomass and a proliferation apparatus therefor.

In order to achieve the above object, the present invention provides a method for culturing microalgae, which comprises culturing microalgae in a nutrient medium to select a plurality of microalgae for objectively measuring the growth rate or growth rate of the microalgae, Wow; The microalgae are introduced into a biological reaction vessel and fed with glycerol as a complex nutrient at a rate of 2 g / L to 5 g / L while supplying air to the biological reaction vessel from the outside, A micro nutrient growth step of growing the microalgae while kneading the microalgae; Obtaining microalgae obtained by microalgae growing through the nutrient growth step of the microalgae and growing microalgae in the biological reaction vessel; .

When the method of growing biomass using glycerol according to the present invention is used, there is an advantage that the amount of microalgae can be further increased irrespective of the kind of microalgae.

In addition, when using the method of growing biomass using glycerol according to the present invention, glycerol, which is obtained as a by-product in the process of converting the obtained microalgae into biomass, is recycled as a complex nutrient for culturing and growing microalgae There is an advantage of not polluting the natural environment.

In addition, when the method of growing biomass using glycerol according to the present invention is used, it is possible to produce a variety of microalgae in a single biomass breeding device, thereby improving the productivity of biomass.

1 is a schematic conceptual diagram of an apparatus 100 for growing a biomass according to the present invention,
2 is a schematic conceptual diagram of a photobioreactor 120 used in the apparatus 100 for growing a biomass according to the present invention,
3 is a schematic conceptual diagram of a light guide plate 126 that can be used in the photobioreactor 120,
4 is a graph showing the production amount of biomass according to the content of glycerol when Chlorella sp. Is used as biomass,
5 is a graph showing the yield of biomass according to the content of glycerol when Nannochloris sp. Is used as biomass,
6 is a graph showing the production amount of biomass according to the content of glycerol when using Botryococcus braunii as biomass.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. It should be understood, however, that the drawings and embodiments are only for the purpose of illustrating the technical concept of the present invention in more detail, and that the technical idea of the present invention is not limited thereto and that various modifications are possible.

The method of growing biomass using glycerol according to the present invention is characterized in that a plurality of microalgae are selected in order to objectively measure the growth rate or growth rate of microalgae and the microalgae obtained by culturing the selected microalgae in a nutrient medium . ≪ / RTI >

The present invention selects microalgae that can be used as a raw material for biomass. When selecting a plurality of microalgae, it is preferable that different microalgae are selected and used. The reason for selecting a plurality of different microalgae for different purposes is to objectively determine whether the same microalgae are added under the same culture conditions and whether the resultant microalgae will have objective and meaningful results to be. The present invention selects Chlorella sp. As a typical biomass of bamboo storage, selects Nannochloris sp. As a representative biomass for seawater, selects Botryococcus braunii as a representative biomass having both bamboo and sea water properties .

When specific microalgae are selected as biomass, the selected microalgae are cultured in BG-11 medium. The culture conditions of the microalgae were maintained at a temperature of 22 ° C ± 2 ° C during the day (8h) overnight (16h), maintaining the pH of the BG-11 culture solution at 7.2 ± 0.3, To < / RTI > 18 days.

Each of the microalgae that have been cultivated is put into a biomass growing apparatus 100 suitable for the biomass growing method according to the present invention.

1 is a schematic conceptual diagram of an apparatus 100 for growing a biomass according to the present invention,

2 is an exemplary conceptual diagram of a photobioreactor 120 used in the biomass propagation apparatus 100,

3 is an exemplary conceptual diagram of a light guide plate 126 that can be used in the photobioreactor 120. As shown in FIG.

The method for growing biomass using glycerol according to the present invention is characterized in that the microalgae are introduced into a biological reaction vessel and fed with air from the outside to the biological reaction vessel so that glycerol as a combined nutrient is fed at a rate of 2 g / L, and the microalgae are grown while being kneaded at room temperature for 10 to 20 days.

In order to efficiently perform the nutrient growth step of the microalgae, it is preferable to utilize the biomass propagation apparatus 100 suitable for carrying out the method of growing the biomass of the present invention. For this, each microalgae that has been cultured in the previous stage is introduced into the biomass propagation apparatus 100, and the biomass propagation apparatus 100 is operated.

The biomass propagation apparatus 100 includes a nutrient supply tank 110 for supplying nutrients from outside to each microalgae cultured in the previous stage and a biomass supply unit 110 for supplying carbon dioxide in the air to the respective microalgae And an air supply tank 112.

The nutrient supply tank 110 stores the nutrient medium and glycerol supplied as the multiple nutrients to the microalgae. The nutrient medium is preferably a BG-11 medium, but not always limited thereto, and a commonly used medium may be used.

The air supply tank 112 supplies carbon dioxide in the air to the microalgae. The air supply vessel 112 may be directly coupled to the photobioreactor 120 associated with each microalgae and may be indirectly coupled to the bioreactor 120 by being coupled to the nutrient supply vessel 110 have. When supplying the air through the air supply reservoir 112, the CO ₂ gas in the air being held dissolved therein an aqueous solution of the photobioreactor 120, the microalgae using a CO ₂ gas, is supplied from the outside It uses light energy to make photosynthesis. Also, the air supplied to the photobioreactor 120 rises upward while causing bubbles, and in the course of the process, the material necessary for growing the microalgae is kneaded.

The nutrient supply unit 110 and the air supply unit 112 are connected to the photobioreactor 120 and supply nutrients, air, carbon dioxide, etc. under the same conditions.

The biomass propagation apparatus 100 includes a plurality of photobioreactors 120 that each of the microalgae can grow by a photosynthesis action and can absorb and grow the complex nutrients supplied from the outside.

It is preferable that the plurality of photobioreactors 120 are determined in accordance with the number of microalgae cultured in the previous stage. For example, if there are two kinds of microalgae cultured in the previous stage, it is preferable that the plurality of photobioreactors 120 are provided in two or more, and when the microalgae are three kinds, Preferably, the number of the photobioreactors 120 is three or more, and when the microalgae are four, the plurality of photobioreactors 120 are preferably four or more, When there are five microalgae, the number of the plurality of photobioreactors 120 is preferably five or more. In the drawings of the present invention, three photobioreactors 120 are exemplarily shown.

The photobioreactor 120 includes a body 122 that stores an aqueous solution containing nutrients of microalgae and microalgae and is made of a predetermined container, An inlet 123 for supplying the nutrient material and air supplied to the body 122 to the body 122, an outlet 124 for discharging the microalgae after the biological treatment to the outside or discharging the slurry to the outside, A light source unit 125 disposed above the light source unit 125 and generating light by an external power source and a light guide plate 125 for uniformly emitting light generated by the light source unit 125 to the inside of the body unit 122 126).

In the photobioreactor 120, glycerol is supplied as a complex nutrient in order to promote the growth of microalgae and to increase the production of microalgae. It is preferable to feed the glycerol at a rate of 2 g / L to 5 g / L. When the amount of glycerol supplied is less than 2 g / L, the growth rate of microalgae and the yield of microalgae are decreased. When the amount of glycerol supplied is more than 5 g / L, the growth of microalgae There is a side where the speed is increased, but it is not preferable because it does not change much compared to the amount of the input. The amount of the glycerol to be supplied may be determined and controlled through the nutrient supply unit 110.

The nutrient medium and the nutrients are introduced into the photobioreactor 120 and the microalgae are introduced into the photobioreactor 120. The light generated from the light source 125 is uniformly dispersed by the light guide plate 126 While slowly kneading for about 17 to 20 days at room temperature. The method of kneading is not particularly limited. The light source 125 generates light using electric energy, and is necessary for the photosynthesis of the microalgae. The light generated by the light source unit 125 is transmitted to the light guide plate 126 disposed below the light source unit 125 and is dispersed evenly inside the body unit 122 by the light guide plate 126. Through such a process, the microalgae grow inside the photobioreactor 120.

The present invention provides glycerol as a complex nutrient from the outside, not only by photosynthesis, but also by the growth of an autotrophic organism as a heterotrophic organism and a heterotrophic organism, in order to promote the growth of microalgae and improve the yield of microalgae It can be said that it pursued growth as a creature at the same time.

In the present invention, it is preferable that the photobioreactor 120 includes a circulation pump 40. The circulation pump 40 continuously performs kneading and circulation of nutrients and microalgae present in the photobioreactor 120. In addition, the photobioreactor 120 may measure the level of the aqueous solution containing the nutrient through the level gauge 30, and determine whether the nutrient is injected from the outside. In addition, it is preferable to provide various measurement sensors 55 so that the temperature, pH, concentration of oxygen or carbon dioxide, concentration of multiple nutrients, and the like of the aqueous solution can be measured. The water gauge 30 and the measurement sensor 55 may be connected to the computer system 50 to automatically perform measurement and control.

The present invention provides a method for producing microalgae, comprising the steps of: discharging microalgae from the microalgae that have undergone the nutrient growth step of the microalgae in the bioreactor (120) and obtaining the microalgae to produce the discharged microalgae as biomass; .

In the present invention, the micro-algae are discharged to the outside through the discharge port (124) in the photobioreactor (120). To this end, the supply of air to the air supply tank 112 is stopped, and the operation of the circulation pump 40 is stopped, and iron sulphate is injected into the inside of the photobioreactor 120. As time elapses, the microalgae cultivated inside the photobioreactor 120 is slurried and settled on the floor. The micro-algae are discharged to the outside through the discharge port 124 and collected as biomass biomass.

Hereinafter, the present invention will be described in more detail with reference to specific examples.

<< Selection and Culture of Microalgae >>

The present invention selected the rectangle Chlorella (Chlorella sp.), And a nano-keulroriseu (Nannochloris. Sp.), And a boat Rio Cocos brownie (Botryococcus. Braunii) as microalgae. The chlorella, nanocholor, and barium cocos brownie were cultivated in BG-11 medium and cultivated in Korean Marine Microalgae Bank (KMMCC, Korea). Culturing conditions were proliferated for 15 days in a thermostat (22 ° C ± 2 ° C) with a cycle of pH (7.2 ± 0.3), day (8h) and night (16h). The components of the BG-11 medium used were as shown in Table 1 below.

BG-11 Medium [mg / L Deionized Water] Component Stock Solution
[g / L dH ₂ O] Quantity used Concentration in
Final Medium [M]   Fe Citrate solution - 1 mL -   Citric acid 6 - 3.12 x 10 ^-5   Ferric ammonium citrate 6 - ~ 3.12 x 10 ^-5 NaNO ₃ - 1.5 g 1.72 x 10 ^-2 K ₂ HPO ₄ .H ₂ O 40 1 mL 1.75 x 10 ^-4 MgSO ₄ .H ₂ O 75 1 mL 3.04 x 10 ^-4 CaCl ₃ .H ₂ O 36 1 mL 2.45 x 10 ^-4 Na ₂ CO ₃ 20 1 mL 1.89 x 10 ^-4 MgNa ₂ EDTA and H ₂ O 1.0 1 mL 2.79 x 10 ^-4   Trace metals solution see following recipe 1 mL
- H ₃ BO ₃ - 2.860 g 4.63 x 10 ^-5 MnCl ₂ and H ₂ O - 1.810 g 9.15 x 10 ^-6 ZnSO ₄ .H ₂ O - 0.220 g 7.65 x 10 ^-7 CuSO ₄ .H ₂ O 79.0 1 mL 3.16 x 10 ^-7 Na ₂ MoO ₂ and H ₂ O - 0.391 g 1.61 x 10 ^-6 Co (NO ₃ ) ₂ .H ₂ O 49.4 1 mL 1.70 x 10 ^-7

<< Biomass Proliferation Device (100) >>

The Chlorella sp., Nannochloris sp., And Botryococcus braunii were cultured in BG-11 medium, respectively, and then added to the biomass growth apparatus 100 to measure the growth activity of the microalgae. Three bio-bioreactors 120 were installed in the biomass propagation apparatus 100, and the micro-algae were introduced into each of the photobioreactors 120 and their initial concentrations were measured. The initial concentration of Chlorella sp. Was 0.357 ± 0.7 g / L, the initial concentration of Nannochloris sp. Was 0.367 ± 0.6 g / L, and the initial concentration of Botryococcus braunii was 0.342 ± 0.7 g / L.

The three photobioreactors 120 were all made in the same shape and the body parts 122 were all 15 L (230 mm (width) * 230 mm (length) * 300 mm (height)) . The photobioreactor 120 was maintained at neutral pH (7.2 ± 0.3), and the progress temperature was 22 ° C. ± 2 ° C., and the cycle of day (8 hours) and night (16 hours) was maintained. Also, the photobioreactor 120 injected 0.5 L of air into the body 112 through the air supply tank 112, and the amount of CO ₂ contained in the air was 0.02 vvm.

A plurality of measurement sensors 55 connected to the computer 50 are coupled to the photobioreactor 120 to measure the state of the aqueous solution of the body part 122. A water level meter 30 is attached to the body part 122, 40) to induce circulation of aqueous solution continuously.

In addition, a V-cut type light guide plate 126 is inserted into the body 122 of the photobioreactor 120 to uniformly transmit light to the entire reactor. The light guide plate 126 made of PMMA (poly-methylmethacrylate) having the best light transmittance was used as a material, and its thickness was 6 mm.

A light source 125 was installed on the light guide plate 126. The light source 125 used an LED lamp and a white color was used. The amount of light used for photosynthesis was ~250 μmol photon / m ² / s. The LED lamps were manufactured in bar form, and the power supplied to the LEDs was supplied by a model FP-60-12 power supply (AD & Lighting, Suwon, Kyonggi-Do, Korea).

In the case where glycerol was not added as a complex nutrient to the microalgae and the glycerol was added at 2 g / L, the glycerol was added at a rate of 5 g / L using the biomass growth apparatus 100 as described above And 10 g / L of glycerol, respectively.

<< Production Increase of Biomass in Photobioreactor 120 >>

Samples of each biomass were sampled at the same time every day while the biomass propagation apparatus 100 according to the present invention was operated and the amount of increase of biomass was calculated through the collected biomass. In other words, 10 mL of each of the samples of Chlorella sp., Nannochloris sp. And Botryococcus braunii were collected at the same time every day, and the increase amount was examined.

The dry weight of microalgae, which can detect the increase in biomass, was measured by filtration of 10 mL of sample with GF / C (Watermann, UK), drying at 105 ° C for 24 hours, and drying mass. The production of biomass was calculated as follows.

P = (X ₁ - X ₀ ) / (t ₁ - t ₀ )

Where P is the production of biomass (g / L / day)

X ₁ and X ₀ denote the amounts of biomass at t ₁ and t _0, respectively.

<< Correlation between the input of glycerol and the production of Chlorella sp. >>

A sample of the inside of the photobioreactor 120 was sampled while operating the biomass propagation apparatus 100 according to the present invention, and the change in the production amount of the Chlorella sp. Was examined. For this, a sample of the Chlorella sp. Was sampled at the same time every day, and the result was calculated by the formula (1).

These experiments were performed on the Chlorella sp., When the compound nutrient glycerol was added at a rate of 2 g / L, when the compound nutrient glycerol was added at the rate of 5 g / L, And nutrient glycerol at a rate of 10 g / L.

As a result, the concentration of glycerol did not affect the amount of biomass until the 5th day of experiment, but after 5 days, the growth of biomass was significantly different according to the concentration of glycerol.

The maximum amount of biomass ( Chlorella sp.) Was 1.31, 1.61, 1.91 and 1.72 g / L, respectively, at concentrations of 0, 2, 5 and 10 g / L of glycerol. Therefore, when the concentration of glycerol was 5 g / L, the amount of biomass of Chlorella sp. Was the best. The Chlorella sp. Showed an increase in biomass of 28.27% when glycerol was added at a rate of 5 g / L compared to the case without glycerol.

The measured results are shown in the table of FIG.

<< Correlation between the input of glycerol and the yield of Nannochloris sp. >>

The same apparatus and method as above were used to sample the inside of the photobioreactor 120, and the change of the production amount of Nannochloris sp. Was examined every day. The resultant value was also calculated by the above equation (1).

Similarly, in the case of Nannochloris sp., When the compound nutrient glycerol was not added at all, when the compound nutrient glycerol was added at the rate of 2 g / L, when the compound nutrient glycerol was added at the rate of 5 g / L, And nutrient glycerol at a rate of 10 g / L.

As a result of the experiment, the effect of glycerol concentration on the amount of biomass was not significant until the 5th day of experiment, but after 5 days, the growth of biomass was significantly different according to the concentration of glycerol.

Similar to Chlorella sp., Nannochloris sp. Showed the best biomass growth at 1.92 g / L at a glycerol concentration of 5 g / L. The Nannochloris sp. Showed at least 37.5% more biomass production when glycerol was used at a rate of 5 g / L compared to no use of glycerol.

The measured results are shown in the diagram of Fig.

<< Correlation between the amount of glycerol and the production of Botryococcus braunii >>

The same apparatus and method as above were used to examine changes in the production amount of Botryococcus braunii while collecting samples of the inside of the photobioreactor 120 on a daily basis. The collected Botryococcus braunii sample was measured, and the result was also calculated by the above formula (1).

Similarly, in the case of Botryococcus braunii , when the compound nutrient glycerol was added at a rate of 2 g / L, the compound nutrient glycerol was added at a rate of 5 g / L, And glycerol at a rate of 10 g / L, respectively.

As a result of the experiment, Botryococcus braunii was isolated from the above-mentioned Chlorella sp. And a biomass proliferation process different from that of Nannochloris sp. The concentration of Botryococcus braunii was 2.47 g / L when the compound nutrient was not added, 2.31 g / L when the compound nutrient was added at 2 g / L, 2.24 g / And 1.87 g / L when 10 g / L of the mixed nutrient was added. Therefore, the Botryococcus braunii showed the best biomass amount at the concentration of 2 g / L glycerol, unlike the above two microalgae. However, there was no significant difference in biomass content between 2 g / L and 5 g / L glycerol. The Botryococcus braunii Showed at least 36.36% more biomass yields when glycerol was used at a rate of 2 g / L, compared to no glycerol at all.

The measured results are shown in the table of FIG.

Although the method and apparatus for growing biomass using glycerol according to the present invention have been described in detail, the present invention is not limited thereto, The range is determined and limited by the range.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope of the present invention.

30: water gauge, 40: circulation pump,
50: computer 55: measuring sensor,
110: nutrient supply tank, 112: air supply tank,
120: photobioreactor, 122: body part,
123: inlet, 124: outlet,
125: light source part, 126: light guide plate,

Claims (1)

A light source unit 125 for generating light by an external power source, and a light source unit 125 for continuously supplying nutrients and microalgae present inside the body unit 122 And a computer system 50 for controlling the measuring sensor 55 and the measuring sensor 55 for measuring the state of the aqueous solution in the body part 122, A photobioreactor (120) that allows algae to further enhance the lipid content after absorption by absorbing the nutrient and is installed in at least one or more parallel fashion;
A nutrient supply tank 110 for supplying nutrients to the body portion; And
An air supply tank 112 for supplying air to the body portion;
1. A method for growing biomass using a biomass propagation apparatus (100) comprising:
The microalgae were selected in order to promote the growth of the microalgae and to improve the production. The microalgae were maintained at a pH of 7.2 ± 0.3 in the culture solution of BG-11, cultured at a period of 16 h during the day (8 h) Culturing the microalgae to obtain microalgae by growing them for 12 days to 18 days while maintaining the temperature at 22 占 폚 2 占 폚;
Feeding the microalgae to the biomass propagation apparatus 100 to grow microalgae;
A step of obtaining a microalga to obtain microalgae grown in the biomass propagation apparatus 100 through a nutrient growth step of the microalgae; Lt; / RTI &gt;
The microalgae use a plurality of microalgae, the plurality of microalgae respectively input into a corresponding plurality of the photobioreactors 120, and air is supplied from the outside to the photobioreactor 120, The microalgae are grown using the biomass propagation apparatus 100 while feeding glycerol as a nutrient at a rate of 2 g / L to 5 g / L and kneading at room temperature for 10 to 20 days,
The air supplied to the photobioreactor is indirectly supplied from the air supply tank 112 through the nutrient supply tank 110,
The light source unit 125 is disposed on the upper portion of the body portion 122 and is provided with a V-cut light guide plate 126 to uniformly diffuse the light generated by the light source unit disposed in the upper portion of the body portion 122, Is provided at a central portion of the body portion (122) so as to uniformly irradiate the fine algae,
PMMA (poly-methylmethacrylate) is used as the material of the light guide plate, the thickness of the light guide plate is 6 mm,
Wherein the microalgae is at least two of the group consisting of Chlorella sp., Nannochloris sp., And Botryococcus braunii. 2. A method for growing biomass using glycerol.

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