KR101499427B1 - Terrimonas ferruginea strain having the ability of promoting growth and flocculation activity of microalgae and uses thereof - Google Patents

Abstract

The present invention relates to a new strain of Terry Monas Peruginia strains having the effect of promoting growth and aggregation of microalgae. Since the strain of the present invention not only promotes growth of the genus Chlorella more than twice, but also improves flocculation efficiency, And are actually applicable to the harvest stage.

Description

Terri Monas Peruvian guinea strain having microalgae growth promoting activity and flocculation activity and its use {Terrimonas ferruginea strain having the ability of promoting growth and flocculation activity of microalgae and uses thereof}

The present invention relates to a novel strain of Terry monas Peruginae having growth promoting activity and flocculation activity of chlorella bulgaris, which is a green alga, and more particularly to a microorganism growth promoting or microbial growth inhibitor comprising Terry monas Peruginia culture medium as an active ingredient, A method for promoting the growth of microalgae including a step of co-culturing a microorganism preparation for coagulating microalgae, a step of co-culturing a Terri Monas Peruvian guinea strain and a microalgae, a step of adding a culture medium of Terri Monas Peruginiya strain to a microalgae culture, A method for improving the flocculation efficiency of the microalgae containing the microorganism, a microorganism preparation for biodiesel production comprising a co-culture solution of Terry Monas Peruvian guinea and chlorella genus as an active ingredient, co-cultivating the Terry Monas Peruvidae strain and chlorella genus strain Room to produce biodiesel It is about the law.

Generally, chlorella is a spherical or elliptical type, and it proliferates very rapidly at 4 to 8 times every 20 hours through the fission of its own cells. It has a very strong vitality and has a large amount of proteins, dietary fiber, vitamins and minerals , It is known as an alkaline food to balance the ion balance of the human body and to have an anti-cancer and antiviral effect, and thus it has been marketed variously as a health supplement food.

As the industrial value of algae has expanded from food to feed and biofuels, cultivation techniques for securing biomes and harvesting techniques for efficient microalgae have become more important. The harvesting of microalgae is a process of separating microalgae from the culture medium, which accounts for 20 ~ 30% of the total biomass production cost, and is one of the important processes in the production of low cost biodiesel.

In the biomass production process using microalgae, the small size of microalgae and the low concentration of the microalgae have difficulties in harvesting biomes, and an efficient and economical method is required (Vandamme, Biotechnology and Bioengineering 2011. vol. 108 (10), 2320-2329).

Generally, coagulation, gravity precipitation, centrifugation, filtration, ultrafiltration, etc. are used for harvesting. Among them, centrifugation is effective and fast, but energy is required, and the filtration method has a disadvantage in that the cost increases because the membrane must be replaced. Aggregation is the most effective way to lower costs and energy (Wyatt, Biotechnlogy and Bioengineering 2012. vol. 109 (2), 493-501).

In the case of algae cells, negative charges are generated. Negative charge prevents algae aggregation in the suspension. Adding metal cations such as calcium or iron to remove or reduce the negative charge of the algae leads to cell aggregation, Larger particles of microalgae allow gravity precipitation, centrifugal recovery, filtration, etc. (Molina, Biotechnlogy Advances 2003. vol. 20 (7-8), 491-515).

Korean Patent No. 351619 discloses 'Fanny Bacillus polyamic acid KCTC 0766BP' for producing a microalgae flocculant, and European Patent Publication No. 02441828 discloses a bioflocculation of algae caused by inactivation of a photoreceptor Algal bioflocculation by inactivation of photoreceptors has been disclosed. However, there is no disclosure of Terry monas Peruginia strains having growth promoting activity and flocculation activity of chlorella bulgaris as in the present invention.

The present invention has been made in view of the above needs, and it is an object of the present invention to provide a method for culturing a microorganism belonging to the genus Terrymonas Peruginia isolated from a microalgae cultured in a stable state through a long period in a laboratory, As a result of carrying out the following growth and flocculation efficiency experiments, the new strain Terry Monas Peruginia strain not only promoted the growth of microalgae more than twice, but also improved the flocculation efficiency, thereby completing the present invention.

In order to solve the above problems, the present invention provides Terrimonas ferruginea strain having microalgae growth promotion and flocculation activity.

In addition, the present invention provides a microorganism preparation for promoting growth of microalgae comprising the culture medium of Terry monas Perugnea strain as an effective ingredient.

The present invention also provides a microorganism preparation for microalgae flocculation comprising the culture medium of Terry monas Perugnea strain as an active ingredient.

The present invention also provides a method for promoting the growth of microalgae comprising co-culturing the Terry Monas Peruviane strain and microalgae.

In addition, the present invention provides a method for enhancing the flocculation efficiency of microalgae, comprising the step of mixing and culturing the microalgae culture broth with the culture solution of the Terry monas Perugnea strain.

The present invention also provides a microorganism preparation for producing biodiesel comprising a co-culture solution of the Terry Monas Peruviane strain and the genus Chlorella as an effective ingredient.

The present invention also provides a method for producing biodiesel by co-culturing a Terry Monas Peruviane strain and a chlorella genus strain.

The new strain Terry Monas Peruginia according to the present invention not only promotes the growth of chlorella genus more than twice, but also improves flocculation efficiency, so that it can be practically applied to microalgae culture and harvesting stages. It can be used for mass production of biomes, Cultivation, harvesting of biomes, and economic cost reduction due to the industrialization process.

Figure 1 shows chlorella growth following co-culture of new strain Terry Monas Peruginia and chlorella bulgaris. (A), chlorella cells according to co-culture (black square) of sterile culture (white circle) of chlorella and Terry Monas Peruvian guinea strain and aseptic chlorella bulgaris strain were measured for 24 days, (B) It shows the number of cells of both strains according to incubation period during co-culture of Monas Peruvian guinea and Chlorella vulgaris.
Fig. 2 shows a schematic diagram for explaining a process for measuring the flocculation activity. (A) shows the coagulation reaction step by step after mixing Chlorella vulgaris and Terimonas Peru Guinea culture (left). After the coagulation reaction, samples were taken at 1 mL in the vicinity of the water surface, and the number of cells was measured using a hemocytometer (Right), and (B) shows the kind of the chlorella culture used for the flocculation activity.
Figure 3 shows the results of DGGE analysis of mixed and incompletely aseptically cultured chlorella. (A) is chlorella hybridized in a long-term laboratory, (B) represents chlorella cultured in an incomplete sterile condition (after FACS treatment), and the band numbers shown in FIG. 3 are the same as those in Table 1.
FIG. 4 shows the flocculation efficiency according to the blending of bacteria isolated from a mixed culture of chlorella bulgaris cultures with chlorella aseptically. E. coli and A8 produced in Bacillus genus known as microbial coagulants as a control group were tested for their flocculation activity in the experiments of Terry Monas Peruginia, Flavobacteria, Hippomonas, Rizobium, Each of the bacteria was cultured for 5 days. The culture solution was filtered with a filter of 0.2 μm and the sterilized chlorella was mixed with the sterilized chlorella. After that (A), (A), (B) (C).
Fig. 5 shows the cohesion rate according to the mixture of Terry Monas Peruvian culture medium and sterile chlorella. A8, Bacillus coagulant, Terry Monas Peruginia culture, and Rizobium medium were mixed with aseptically cultivated chlorella, and the chlorella coagulated at 0 day (upper) and 1 day (lower), respectively .
FIG. 6 shows the cohesion rate according to the mixing of bacterial cells isolated from chlorella culture solution of mixed state and sterile chlorella cells. E. coli and A8 produced in Bacillus genus known as microbial coagulants as a control group were tested for their flocculation activity in the experiment of Terry Monas Peruginia, Flavobacterium, Hippoponas, Each bacterial cell was incubated for 5 days with 3 × 10 ⁵ cells / ml of bacterial cells and 1 × 10 ⁶ cells / ml of aseptic chlorella Showed agglutinating activity after 8 days of mixing.
FIG. 7 shows the results of comparing the flocculation efficiencies of the aseptic and hybrid chlorella cultures according to the cation (Ca, Fe) concentration (mM). (A) an aseptic chlorella culture, (B) a hybrid chlorella culture
Fig. 8 is a microphotograph of a sample of sterile and hybrid chlorella culture used for coagulation efficiency measurement. (A) an aseptic chlorella culture, (B) a hybrid chlorella culture
FIG. 9 shows the zeta potential measurement results of the aseptic and mixed chlorella culture according to the pH and the cation concentration. (A) an aseptic chlorella culture, (B) a hybrid chlorella culture
FIG. 10 shows the result of mixing and coagulation efficiency between microalgae cells of an aseptic culture medium and a mixed culture medium in which microalgae were removed.

In order to achieve the object of the present invention, the present invention relates to a method for producing a microorganism having Terrigonas ferruginea .

The new strain of the present invention was identified as Terry Monas Peruginias as a result of the identification of microorganisms isolated from microalgae cultures cultured in a stable state after a long passage in a laboratory through DGGE analysis as a molecular biological method, (Accession No .: KCTC 12269BP) on August 23, 2012 at the Korea Research Institute of Bioscience and Biotechnology.

In one embodiment of the invention, the microalgae are selected from the group consisting of Chlorella sp., Spirullina sp., Chlamydomonas sp., Chlorococcum sp., Dunaliella sp., Ellipsoidion (Ellipsoidion) in nano claw drop system (Nannochloropsis), A neo keulroriseu (Neochloris), A Pavlova (Pavlova), An L ohdak tilrum (Phaeodactylum) genus, three or four des mousse (Scenedesmus) in, in tetra cell Miss (Tetraselmis) , isopropyl Chris (Isochryis) and the like in, preferably may lie chlorella (chlorella), more preferably, chlorella vulgaris (chlorella vulgaris ). < / RTI >

In addition, the present invention provides a microorganism preparation for promoting growth of microalgae comprising the culture medium of Terry monas Perugnea strain as an effective ingredient.

The microbial formulation for promoting microalgae growth according to the present invention may be prepared as a solution, a powder, or a suspension, but is not limited thereto.

The present invention also provides a microorganism preparation for microalgae flocculation comprising the culture medium of Terry monas Perugnea strain as an active ingredient.

The microorganism preparation for microalgae flocculation according to the present invention may be prepared as a solution, a powder or a suspension, but is not limited thereto.

The present invention also provides a method for promoting the growth of microalgae comprising co-culturing the Terry Monas Peruviane strain and microalgae.

In the step of co-culturing microalgae and bacteria according to an embodiment of the present invention, the number of inoculated cells is 0.5 × 10 ⁶ to 1.5 × 10 ⁶ cells / ml of chlorella bulgaris And Terry Monas Peruvian guinea strain may be 2 x 10 ⁵ - 4 x 10 ⁵ cells / ml, preferably 1 x 10 ⁶ cells / ml And 3 x 10 < ⁵ > cells / ml But is not limited thereto.

The method for co-culturing the strain may be any method known in the art and is not particularly limited to a specific method. For example, the co-culture of the strain may be cocultured for 24 days at 25 ° C, 100 rpm, 100 μmol m ^-2 s ^-1 light, but is not limited thereto.

In addition, the present invention provides a method for enhancing the flocculation efficiency of microalgae, comprising the step of mixing and culturing the microalgae culture broth with the culture solution of the Terry monas Perugnea strain.

In the method according to one embodiment of the present invention, when the bacterial culture broth and the aseptic chlorella genus are mixed, the Terry monascus Perugnea strain is cultivated for 5 days, then filtered with a 0.2 mu m filter and the sterilized chlorella 6 X 10 < ⁶ > cells / ml may be mixed and cultured for 0, 1, and 3 days, preferably 0 to 1 day, but the present invention is not limited thereto.

In the method according to an embodiment of the present invention, Ca ^{2 +} or Fe ^{3 +} ions may be added as an auxiliary coagulant of the mixed culture liquid in order to improve the flocculation efficiency of microalgae, but the present invention is not limited thereto.

In the method according to one embodiment of the present invention, the pH of the mixed culture liquid may be 3 to 12, preferably 10 to 12, and most preferably, May be pH 11 but is not limited thereto.

The present invention also provides a microorganism preparation for producing biodiesel comprising a co-culture solution of the Terry Monas Peruviane strain and the genus Chlorella as an effective ingredient.

The microorganism preparation for producing biodiesel of the present invention can be produced by using a co-culture solution of a Terry Monas Peruvinia strain and a chlorella genus strain as an effective ingredient. The microbial formulation for producing biodiesel according to the present invention can be prepared by solution, powder or suspension, but is not limited thereto.

Also, the present invention provides a method for producing a polyunsaturated fatty acid, which comprises co-culturing the Terry Monas Peruviane strain and the chlorella genus to produce a fatty acid; And converting the produced fatty acid into biodiesel. The present invention also provides a method for producing biodiesel.

The method for converting the fatty acid produced by the strain of the present invention into biodiesel may be any method known in the art and is not particularly limited to a specific method.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Materials and methods

1. Chlorella Bulgarian  Aseptic culture and Mixed culture ( Xenic culture )

Chlorella vulgaris used JQ664295 and cultured in BG11 medium obtained from livestock wastewater from Gongju province. Chlorella vulgaris was cultured in a 1 L flask in 300 mL BG11 medium. The culture conditions were incubated at 25 ° C at 100 rpm and 100 μmol m ^-2 s ^-1 light condition. Aseptic chlorella bulgurris colonies were harvested from livestock wastewater, sonicated and separated by Fluorescence-Activated Cell Sorting (FACS). Chlorella hybridized in a long-term laboratory was maintained on BG11 medium every 3 months for 14 days by subculture.

2. Aseptic ( Axenic ) Chlorella in the state Bulgarian and  Co-culture with isolated bacteria Co - cultivation )

Put 50ml BG11 medium in 250ml flasks cultured chlorella to aseptic (axenic) condition vulgaris (Chlorella The cells were cultured in BG11 medium supplemented with 1 × 10 ⁶ cells / ml and 200 ppm of glucose for 5 days. The cells were counted by DAPI (4 ', 6-diamidino-2-phenylindole) 10 < ⁵ > cells / ml. After inoculation, the cells were cultured for 24 days at 25 ℃, 100 rpm, and 100m ^{- 2} s ^{- 1} . The number of microalgae cells was counted using a hemocytometer, and the DAPI staining method was used for the cells.

3. Coagulation activity and pH Titration of

The pH range of the sterile and hybrid chlorella culture after incubation is 8-9. After incubation, the culture was titrated to a concentration of 1 × 10 ⁶ cells / ml and the pH was adjusted to 3, 5, 7, 9, 11 with 1N NaOH and 1N HCl. 50 ml of the chlorella culture solution was mixed at 300 rpm for 30 seconds and then mixed at 100 rpm for 1 minute. CaCl ₂ , a cationic coagulant, was added and mixed at 300 rpm for 30 seconds and then at 100 rpm for 1 minute. Next, FeCl ₃ was added to the culture, followed by mixing at 300 rpm for 30 seconds and mixing at 100 rpm for 1 minute. The supernatant of the culture broth was taken 1ml each after 2 minutes. Cell numbers were determined microscopically using a C-chip hemocytometer (Figure 2). The coagulation activity was calculated by the following equation.

Coagulation activity (%) = (1-cell number after flocculation / cell number before flocculation) × 100

4. Mixing and coagulation efficiencies of bacterial filtered cultures and aseptic chlorella cells

In a 250 ml flask, BG11 medium supplemented with 200 ppm of glucose was added to Terrimonas ferruginea), Flavobacterium genus (Flavobacterium sp.), Hippo Pseudomonas genus (Hyphomonas sp.), Rizzo Away in (Rhizobium sp.), the A8 produced by Bacillus known as E. coli and microbial coagulants were cultured for five days. Culture solution by a 0.2㎛ (Millipore Express Plus membrane filter) contained in the culture solution to remove the bacterial cells to filter a sample of 50ml of Chlorella ⁶ × 10 6 cells / ml in a sterile 100ml beaker in parallel to high-speed stirring and low-speed stirring 10mM CaCl ₂ And 0.26 mM FeCl ₃ as a cation flocculant, the flocculation activity was confirmed immediately after 1 day and 3 days. Cell counts were measured using a C-chip hemocytometer under a microscope and the coagulation activity was calculated.

5. DGGE  Analysis and sequencing

Four different samples of chlorella hybridized in a long-term laboratory for genomic DNA extraction, incomplete aseptic culture (after FACS treatment), supernatant obtained after centrifugation of mixed chlorella culture, and filtered culture were taken. For DGGE (denaturing gradient gel electrophoresis) analysis, 16S rRNA The nucleotide sequence was partially amplified. PCR was carried out in two sets and the primers were GC-clamp 341F (5'CCT ACG GGA GGC AGC AG 3 '; SEQ ID NO: 1), GC-clamp (5'CGC CCG CCG CGC GCG GCG GGC GGG GCG GGG GCA CGG GGG G 3 '; SEQ ID NO: 2) and 786R (5' CTA CCA GGG TAT CTA ATC 3 '; SEQ ID NO: 3). DGGE was performed using a Dcode ™ system (Bio-Rad Laboratories, USA) and the PCR products were electrophoresed in 8% (w / v) polyacrylamide (37.5: 1 acrylamide / bisacrylamide) ). Electrophoresis was performed in 1 x TAE (40 mM Tris, 20 mM acetate, 1 mM EDTA pH 7.4) at 60 [deg.] C and 60 V for 18 hours. The gel was stained with EtBr (ethidium bromide) (0.2 ㎍ / ml, 1 × TAE) for 15 minutes and desalted for 5 minutes with DW (deionized water) and confirmed using KODAK Gel Logic 100 Imaging System. Each DGGE band was extracted from the gel and amplified using it as a PCR template. The nucleotide sequence of the amplified DNA was summarized by SeqMan software (DNA STAR, Madison, Wis.), And the nucleotide sequence was identified using Blast (Basic Local Alignment Search Tool) of the database of the gene bank (Table 2). The resulting gene was registered in the gene bank database (accession number JX270632 to JX270636).

6. Mixed  Culture solution FACS  process

Incomplete aseptic chlorella culture is a hybrid culture in which cells are partially isolated using BD FACSAria cell sorter (Becton Dickinson, USA). Flow-cytometry and cell sorting experiments are described in Doan, Biomass and Bioenergy, 2011. 35 (7), 2534-2544) and Sensena (Sensena, European Journal of Phycology, 1993. 28 -97). ≪ / RTI > Forward scatter width profiles were used to separate microalgae and bacterial cells. The microalgae were isolated through a 695/40 nm per CP-Cy5.5 filter. These cells are classified into 15 mL sterile tubes for centrifugation. Clones were cultured in which one cell was classified into a tube. Tubes were opened prior to sorting and immediately closed as soon as they were sorted, and this separation was performed at least twice. This tube is continuous brightness 100μ㏖ m ^-2 s at 25 ℃ ^- were incubated with the ^1.

Example  One: Mixed  In the chlorella culture, the bacterial community community ) Isolation and identification

To identify bacterial communities in hybrids of chlorella cultures, DGGE (denaturing gradient gel electrophoresis) was performed using mixed cultures to compare the bacterial diversity from the hybrid cultures. Flavobacterium sp, accession number JX270632, Terrimonas spp. ferruginea , accession number JX270633), Sphingobacterium sp. accession number Jx270634, Rhizobium sp. accession number Jx270635 and Hyphomonas sp. accession number JX270636 were isolated Table 1).

Example  2: Terry Monas Peru Guinea  Chlorella of strain Bulgarian and  Promoting growth of chlorella in co-culture

Bacteria isolated from chlorella and microalgae cultures cultured aseptically were co-cultured. Chlorella vulgaris cultured in sterile condition with Terry Monas Peruginia isolated for 24 days showed that the number of cells co-cultured was as low as 4.16 × 10 ⁷ cells / ml in case of chlorella cultured aseptically, 9.06 × 10 ⁷ cells / showed more than twice the rate in ml (Fig. 1A), when viewed as the number of chlorella vulgaris and bacterial cells increased gradually according to the culture period is a terry Pseudomonas Peru Guinea strain isolated from micro-algae culture chlorella vulgaris and mutualism (Fig. 1B) by maintaining the relationship of mutualism (Fig. 1B).

Example  3: Mixed  Of cultured chlorella pH  Increase of flocculation activity with increase

In the case of chlorella cultured in aseptic condition, the pH does not increase the flocculation activity, but in the case of mixed chlorella, flocculation activity increases with increasing pH. These results indicate that the bacterial community directly affects the flocculation activity of chlorella, and that the higher the pH, the higher the flocculation efficiency. The hybrids cultured chlorella showed high flocculation activity of 94% at pH 11 (Table 2).

Example  4: Terry Monas Peru Guinea  Comparison of Coagulation Efficiency of Chlorella and Other Bacterial Cultures to Chlorella

E. coli and A8 produced in the genus Bacillus, which are known as microbial coagulants, are used as the coagulation efficiency (A) and the coagulation efficiency Respectively. Each of the bacteria was cultured for 5 days, and the culture solution obtained by removing the bacterial cells with a filter of 0.2 탆 (Millipore Express Plus membrane filter) was inoculated with 6 × 10 ⁶ cells / ml of aseptic chlorella immediately (FIGS. 4A and 5) After 1 day (FIG. 4B and FIG. 5), the coagulation efficiency after 3 days (FIG. 4C) was confirmed. When the coagulation efficiency was confirmed immediately after mixing, the coagulation efficiency was very high in the bacteria other than the Rizobium, but the coagulation efficiency was decreased as time elapsed (FIG. 4A-C). In the coagulation efficiency measurement after 3 days, it was confirmed that Terry monas Peruginia strains showed the highest coagulation efficiency of 60% or more (Fig. 4C) as compared with other bacteria (Fig. 4C) Appears to help improve the coagulation effect.

The cells were harvested from Terry Monas Peruginia, Flavobacterium, Hippomonas, and Rizobium spp. Isolated from chlorella culture broth and mixed with sterile chlorella culture for 8 days. Respectively. The rest of the species except Terry Monas Perugiania showed a coagulation efficiency of 20%, whereas Terry Monas Perugiania showed 2.5 times higher coagulation efficiency (Fig. 6). Therefore, it can be seen that co-cultivation of Terry Monas Peruvian guinea cells with chlorella effectively improves the flocculation efficiency of chlorella.

Example  5. Cationic agent  Aseptic, Mixed  Comparison of Coagulation Efficiency of Chlorella Culture Solution

Experiments were carried out on cationic (Ca, Fe) concentrations to obtain optimum microalgae flocculation efficiency in hybrids of chlorella culture containing sterile chlorella bulgurris, terrimonas ferruginea and other microalgae interacting bacteria . At optimum pH 11, the coagulation efficiency of the aseptic chlorella culture (A) was close to 0%, but the bacterial hybridization broth (B) showed a coagulation efficiency of 90% or more at the maximum. In the case of chlorella cultured in aseptic condition, it can be seen that there is no influence on flocculation activity even when the concentration of the cationic agent is changed as well as the pH (Fig. 7).

Example  6. Aseptic, Mixed  Microscopy of Chlorella culture samples

In order to visually confirm how the bacterial community is acting on microalgae aggregation, the coagulation experiments were completed and the specimens were observed under a microscope. According to the microscopic results, microalgae floc was formed in both aseptic (A) and mixed (B) chlorella, but the size was much larger in the mixed chlorella culture. As a result, it was found that the effect of Terrimonas ferruginea bacteria and its metabolites accelerated the aggregation of microalgae to reach a condition easy to precipitate (Fig. 8).

Example  7. pH Wow Cationic agent  Aseptic, Mixed  The zeta potential of the chlorella culture ( zeta potential ) Measure

The zeta potential was measured by varying the concentration of the cationic agent in the aseptic (A), mixed (B) chlorella culture without pH control and in the experimental solution adjusted to the optimum pH of 11 (the zeta potential was dispersed The electric potential difference between the particles and the surrounding culture liquid can be obtained, and the stabilized state of the particles can be known by this value). The zeta potential is close to zero, which indicates that it forms a perfect flock. The zeta potential value is close to zero at pH 11 in the aseptic culture. On the other hand, the value of close to 15 in the mixed culture indicates that it is insufficient to form the flock, but the fact that it has a flocculation efficiency of more than 90% explains that the role of bacteria plays an important role in flux formation (Fig. 9).

Example  8. Confirmation of mixing and coagulation efficiency of microalgae cells and microalgae-free mixed culture of sterile culture medium

Bacterial cell broth containing weak bacterial cells obtained by centrifugation and filtered bacterial cells that have been filtered have been removed in order to carry out an experiment on whether or not the bacteria directly affect the aggregation of microalgae. A supernatant of two kinds of mixed culture was obtained. Herein, only the microalgae cells were obtained from the aseptic culture, and the resultant mixture was mixed with each supernatant. Then, the flocculation efficiency was measured immediately after the mixing at the optimum pH of 11 and 24 hours after the mixing (FIG. 10). Immediately after mixing, 83% of the bacterial cell broth and 55% of the filtered broth showed bacterial cell aggregation efficiency. After 24 hours, the coagulation efficiency was not significantly different between the aseptic culture and the mixed culture. However, in both cultures of bacterial cell broth and filtered broth, there was a tendency to increase within 10% It looked. The efficiency of coagulation of the filtered broth with bacterial cells is about 30% lower than that of the bacterial cell broth, but the efficiency is 50% higher than that of the aseptic culture. The following results show that both bacterial cells and their metabolites can be factors influencing microalgae aggregation.

Korea Biotechnology Research Institute KCTC12269BP 20120823

<110> Korea Research Institute of Bioscience and Biotechnology <120> Terrimonas ferruginea strain having the ability of promoting          growth and flocculation activity of microalgae and uses thereof <130> PN13287 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 1 cctacgggag gcagcag 17 <210> 2 <211> 40 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 cgcccgccgc gcgcggcggg cggggcgggg gcacgggggg 40 <210> 3 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 ctaccagggt atctaatc 18

Claims (11)

Terrimonas ferruginea strain (KCTC 12269BP) with microalgae growth promoting and flocculating activity. The strain according to claim 1, wherein the microalgae is a genus of Chlorella . The method of claim 2, wherein the chlorella is a strain which is characterized in that the Chlorella vulgaris (Chlorella vulgaris). A microorganism preparation for promoting growth of microalgae in Chlorella containing the culture medium of Terry Monas Peruviane strain of claim 1 as an active ingredient. A microorganism preparation for microalgae coagulation in Chlorella containing the culture medium of Terry Monas Peruviane strain of claim 1 as an active ingredient. A method for promoting the growth of Chlorella microalgae comprising the step of co-culturing the Terry Monas Peruvian guinea strain and the Chlorella microalgae of claim 1. A method for enhancing the flocculation efficiency of microalgae of the genus Chlorella including the step of culturing the microalgae culture medium of Chlorella microorganism by adding the culture medium of Terry Monas Perugnea strain of claim 1 and mixing. [8] The method according to claim 7, wherein Ca2 + or Fe3 + ions are added as an auxiliary coagulant to the culture medium of Terry Monas Peruviane strain, to improve the flocculation efficiency of microalgae of the genus Chlorella. [8] The method according to claim 7, wherein the pH of the mixed culture liquid is 10-12. A microorganism preparation for producing biodiesel comprising the co-culture solution of the Terry Monas Peruviane strain and the chlorella genus strain of claim 1 as an active ingredient. Culturing the Terry Monas Peruviane strain and the chlorella genus strain of claim 1 to produce a fatty acid; And
And converting the produced fatty acid to biodiesel.

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