KR101427979B1 - Method for Treating Mixed Seaweed Wastes Using Bacillus alcalophilus - Google Patents

Abstract

The present invention relates to a method for treating mixed seaweed waste using bacillus alcalophilus KACC91705P having seaweed derived polysaccharide decomposition ability and protein decomposition ability. According to the present invention, reduced monosaccharide like galactose and glucose and oligosaccharide can be produced from various kinds of seaweed waste compounds or byproducts which are difficult to be treated and is produced during a seaweed treating process or during extraction, to not only have an effect of being recycled for the production of biomolecule or bioethanol, but also have an effect of solving environmental problems by effectively treating mixed seaweed waste.

Description

Technical Field [0001] The present invention relates to a method for treating mixed seaweed waste using Bacillus alcalophilus,

The present invention relates to a method for treating a mixed seaweed waste using Bacillus Calphilys KACC91705P having decomposition ability and protein decomposition ability of seaweed-derived polysaccharide.

So far, seaweed polysaccharides have been extracted from marine algae through processing methods such as alkaline, acid or enzyme treatment, and they have been widely used industrially as food, cosmetic additives, and health food resources. Now, they are used as resources for bioethanol production It is expanding. However, the amount of algae required to produce bioethanol is 12.5 million tons, while the amount of algae that Korea can produce is limited to 1 million tons. Therefore, it is difficult to utilize seaweed, which is widely used in industry such as food, for the development of bioenergy. Therefore, bio-energy and bioactive materials are to be developed with the use of fish wastes generated during processing and extraction processes. However, it is difficult to use as a substrate for biofuel production due to the complicated structure of seaweeds itself, which is difficult to decompose, and aquatic wastes (harmful algae) generated during the processing process and extraction, and the waste is not separated from the process There is a problem that by-products and wastes generated from various seaweeds are mixed. In particular, Korea is obliged to comply with the '96 Protocol adopted by the London Convention, which regulates waste dumping as a member of the OECD. Therefore, it is not possible to exclude the possibility of a dispute arising from disposal of seaweed after 2013 . Accordingly, it is possible to grasp the problems that these problems and various wastes and by-products generated in the process of seaweed processing or during the extraction are not separated in the treatment process but the various seaweeds are mixed, Patented microorganisms decomposing red algae, agar, and carrageenan purely separated from soil, tidal flats, and internal organs of animals, not by decomposing seaweeds by chemical (alkali, acid) or physical (heat) , It is possible to decompose mixed harmful algae and by-products, which are environmental treatment problems, by using the characteristics of synthesizing other complex enzymes. In addition, monosaccharides, galactose and glucose produced as a result of degradation are used not only for the production of ethanol through the yeast fermentation process, but also for degradation products such as peanut, sugars, oligosaccharides, It can be developed as a useful physiologically active substance such as mold.

So far, microorganisms known to degrade agar or carrageenan by enzymatic method include agarolorans (agarivorans sp.), Which has a heat-resistant beta- agarase for producing agar oligosaccharide in a high-temperature nano-particle state . Thalassomonas agarivorans (Thalassomonas sp. SL-5, KCCM 10790P) (Korean Patent No. 0794593) having beta- agarase for producing agar oligosaccharide, Saccharophagus degradans 2-40 ( Saccharophagus sp. AG21, KFCC 11416P) (Korea patent application 2008-0038244) having both alpha- agarase and beta- agarase for producing galactose and maintaining excellent agar resolving ability even at 40 ° C or higher And it is possible to produce various agar oligosaccharides which are high value-added products using agar as a raw material, Glaciecola mesophila AJ548479 ( Glaciecola sp. SL-12, KCCM 10945P) (Korea Patent No. 1,072,503) which can produce beta-agarase capable of producing high-value-added neoagarooligosaccharide has been reported.

As a microorganism having carrageenan resolution, Pseudomonas sp. (Council of Scientific and Industrial Research, 2008. Republic of Korea Application No. 10-2008-7026331), production of carrageenan and production of carrageenan using MTCC 5261 (JP 1006656, JP 2000116376) have been reported. However, the development of microorganisms capable of degrading carrageenan has not been developed yet.

Microorganisms known to degrade cellulose or laminarin by an enzymatic method include microorganisms having cellulolytic ability, such as cellulose degrading enzyme, a new microorganism producing endoglucanase, Xanthomonas sp. EC102 (Korean Patent No. 1,132,396), Thermus caldophilus GK24 which produces a cellulase for the production of short-lived beta-glucan including cellulose 2 and cellobiose through hydrolysis of beta-glucan No. 1,137,020), Stereum hirsutum SKU512 (KCCM 10982P) (Korea Patent Publication No. 2011-0042397), Trametes hirsuta SKU804 (KCCM 10960P) (Korea Patent Publication No. 2010-0042547 ) And Schizophyllum commune KMJ820 (KACC 93084P) (Konkuk University-Industry-Academic Collaboration Foundation 2012. Use of skilfull rum kojic and saccharification to produce cellulase, registration number: 10-1135178-0000), and xylanase and low temperature of 10 ° C Of the maximum activity according to the change of temperature, and Bacillus licheniformis DK4 which secretes easily the cellulase into the feed additive for decomposition enhancement 2 (KACC 91410P) (Korean Patent No. 1,062,309) have been reported.

Microbes decomposing kelp and seaweed, Microbacterium sp. Although A2203 (KACC 91096) (Korean Patent No. 625,299) has been reported, development and reports of various seaweeds, seaweed polysaccharides, and laminarin-degrading microorganisms have not been developed yet.

However, there are numerous strains that produce one enzyme that degrades polysaccharides. However, there are many strains producing agar and carrageenan, cellulose, laminarin, and other seaweed polysaccharides, as well as other algae (green algae and brown algae) as well as red algae containing agar and carrageenan. It has not yet been developed for microorganisms that directly degrade alginate, fucoidan, and protein, and has not been reported so far. When a mixture of enzymes synthesizing the polysaccharide is prepared, Mutual action and optimum condition, and treatment efficiency by culture may be lowered. However, the strains of the present invention can complement these disadvantages by synthesizing enzymes capable of decomposing various types of seaweed polysaccharides from a single microorganism.

Accordingly, the present inventors have made intensive efforts to develop a strain capable of effectively treating a harmful algae occurring in the process of processing seaweed or during the extraction process. As a result, it has been found that seaweed polysaccharides such as agar and carrageenan, cellulose and laminarin, It has been confirmed that Bacillus alcalophilus having excellent ability to decompose alginic acid, fucoidan and protein can be isolated and the mixed seaweed waste can be effectively treated by using the strain. Thus, the present invention has been completed.

It is an object of the present invention to provide a method for decomposing algae using a strain having the ability to decompose polysaccharides derived from algae and to decompose proteins.

Another object of the present invention is to provide a method for treating mixed seaweed waste using a strain having the ability to decompose polysaccharides derived from seaweeds and to decompose proteins.

In order to achieve the above object, the present invention provides a method for producing a reducing sugar by decomposing a seaweed or seaweed polysaccharide, which comprises using Bacillus alcalophila KACC91705P.

Further, the present invention provides a method for treating a mixed seaweed waste characterized by using Bacillus arcuarophilus KACC91705P.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to produce reduced monosaccharides such as galactose, glucose, and oligosaccharides from various kinds of disused seaweed mixture or by-products generated in the process of seaweed processing or difficult to be generated during extraction, It is effective not only to effectively treat the mixed seaweed waste but also to improve the environmental problem.

FIG. 1 shows the results of confirming the resolving power of the SYR1 strain against the Kim powder (A), agar (B) and carrageenan (C).
Fig. 2 shows the results of confirming the resolution of the strain SYR1 against parasitism (A) and kelp (B).
Fig. 3 shows the results of confirming the resolving ability of the SYR1 strain against cellulose (A) and laminarin (B).
Fig. 4 is a graph comparing the results obtained by confirming the resolving power against the red algae (Kim), green alga (blue), and brown algae (kelp) of the strain SYR1.
FIG. 5 is a comparison of the results obtained by confirming the resolving ability of seaweed polysaccharide (agar, carrageenan, cellulose, laminarin, alginic acid, fucoidan) of strain SYR1.
FIG. 6 is a graph showing the results of decomposition of Kim powder in SYR1 strain [pH (); Cell density (●); Total reducing sugars (■)].
FIG. 7 is a graph showing the results of parsley (A) and kelp (B) powder degradation of SYR1 strain [pH (); Cell density (●); Total reducing sugars (■)].
8 is a graph showing the results of agar (A) and carrageenan (B) degradation of strain SYR1 [pH (); Cell density (●); Total reducing sugars (■)].
9 is a graph showing the results of degradation of cellulosic (A) and laminarin (B) of strain SYR1 [pH (); Cell density (●); Total reducing sugars (■)].
Fig. 10 shows the results of confirming the resolving power of the cellulose (A) and laminarin (B) of the strain SYR1 under optimal conditions.
Fig. 11 shows the clear zone by proteolysis after the strain SYR1 was cultured in a skim milk solid plate culture medium.

In one aspect, the present invention relates to a method for producing a reducing sugar by degrading a seaweed or seaweed-derived polysaccharide, which comprises using Bacillus alcalophila KACC91705P.

The Bacillus alcalophilus strain KACC91705P of the present invention is a microorganism that directly degrades red algae containing agar and carrageenan, and also produces cellulase and laminaracetate. In addition to agar and carrageenan, it also contains cellulose, laminarin, alginic acid, fucoidan, As a new strain to degrade together, it is possible to treat the problems of mixed by-products and wastes generated in seaweed processing and extraction processes, and to produce biofuels and bioactive materials.

To date, no microorganisms have been disclosed which directly decompose seaweed polysaccharides such as agar, carrageenan, cellulose, laminarin, alginic acid, fucoidan, and protein together with other seaweeds (algae, brown algae).

In the present invention, the seaweed may be red algae, green algae or brown algae, and the seaweed-derived polysaccharide may be selected from the group consisting of agar, carrageenan, laminarin, alginic acid, fucoidan and cellulose.

In the present invention, the microorganisms capable of efficiently degrading agar and carrageenan, which are waste seaweed polysaccharides produced in the red algae processing step and the extraction step, are separated from samples collected in various environments such as soil, seawater and tidal flat, It was confirmed that the carotenoids KACC91705P degrades cellulose and laminarin, alginic acid, fucoidan and protein in addition to agar and carrageenan. In addition, when the green alga (cellulose) and brown alga (laminarin) were decomposed by the microorganisms, it was confirmed that glucose and oligosaccharide reducing sugars were produced as degradation products.

Bacillus alcalophilus KACC91705P of the present invention is capable of degrading algae containing lignin and laminarin containing celluloses in addition to red algae containing agar and carrageenan. In one embodiment, in a solid medium containing parasites and sea tangle, Alkalophilus KACC91705P was cultivated and it was confirmed through the Rugol solution to form a transparent ring due to decomposition of kelp and blue seaweed.

In another aspect, the present invention relates to a method of treating seaweed waste characterized by the use of Bacillus arcuarophilus KACC91705P.

In the present invention, the seaweed waste may be a seaweed waste, a fish waste, or a mixed waste of seaweed waste and fish waste.

Bacillus alcalophilus KACC91705P of the present invention is capable of degrading cellulose and laminarin, alginic acid, fucoidan and protein in addition to agar and carrageenan. In one embodiment, each of the solid medium containing laminarin, alginic acid and fucoidan, , Bacillus alcalophilus KACC91705P was cultivated and it was confirmed through the Rugol solution that a transparent ring was formed by the decomposition of cellulose and laminarin, alginic acid and fucoidan. In addition, Bacillus alcalophilus KACC91705P was cultured in a solid medium containing skim milk, and it was confirmed that a transparent ring was formed by proteolysis.

In addition to agar and carrageenan contained in red algae, Bacillus alcalophilus KACC91705P of the present invention decomposes laminarin, alginic acid and fucoidan contained in cellulose and brown algae contained in green algae and produces reducing sugars of galactose, glucose and oligosaccharides as degradation products . These reducing sugars can be used for bioethanol production and physiologically active substances. As a result, in addition to the production of bioethanol and physiologically active substances using harmful algae mixed with red algae, green algae and brown algae, Staphylococcus aureus can be very useful. In addition, it has the ability to decompose the protein, so it will be easy to treat mixed seaweed waste mixed with fish waste in the future.

Hereinafter, the present invention will be described in more detail by way of examples. It will be apparent to those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited to these embodiments.

Example 1. Pure isolation of strains having excellent seaweed degrading ability

Bacteria that effectively degrade agar and carrageenan were isolated from samples taken from the soil, tidal flats, and septic tanks of marine animals.

Samples collected from soil, tidal flats and marine animals were inoculated into a Kim supplement medium (1.5 g of Kimchi itself, Sea water, pH 7.42) and incubated for 14 days at 37 ° C with 180 rpm shaking or in a Kim powder medium _{/ L, NH 4 Cl 1 g} / L, Tap water, and then inoculated to pH 6.8) shaking culture at 7 days 37 ℃, 180 rpm, 1% laver powder medium _{(NH 4 Cl 1 g / L} , Tap water , pH 6.8) was plated on a solid medium supplemented with 1.5% agar. The cultured microorganisms were continuously plated on a new solid medium until the microorganisms were separated into pure colonies, and the strains were finally purified. The isolated pure strains were subcultured every 2 weeks with storage at 4 ℃.

Bromocresol Purple (0.5 g / L) and 1.5 (1.5 g / L) were added to a Kim powder medium (Kim powder 10 g / L, NH ₄ Cl 1 g / L, Tap water, pH 6.8) to screen for useful microorganisms in the separated pure strains. % Agar was added to the solid medium.

As a result, it was confirmed that the strain SYR1 showed changes in medium color depending on the pH change in the agar medium containing the indicator powder.

In order to investigate the resolving ability of the above strain against Kim, agar and carrageenan, the SYR1 strain was inoculated into 5 ml of the Kim powder liquid medium and incubated at 37 DEG C and 180 rpm for 1 to 2 days with shaking. 10 [ The cells were dropped on a plate medium, dried, and then cultured at 37 ° C for 4 days.

After confirming that each strain formed a colony on the solid medium, 10 ml of Lugol's solution (KI 10 g / L, I 5 g / L) was added to the Kimpard plate culture medium and stained for 5 minutes. , And then the clear zone due to the decomposition of Kim was confirmed. The resolving ability of the strain was confirmed, and the resolution of agar and carrageenan was confirmed in the same manner (FIG. 1).

As a result, it was confirmed that the SYR1 strain showed high resolution in each of the kim powder, agar and carrageenan solid plate culture media. According to various pH and temperature conditions, optimal culture and resolution conditions of the strain were pH 7.5-8 (pH 7.8 ) And a temperature of 30 ° C.

The 16S-rRNA sequence of the isolated SYR1 strain was analyzed and performed. The chromosomal DNA of each strain was isolated using an AccuPrep chromosome DNA extraction kit (Bioneer, Korea) and PCR was performed using the following primers (SEQ ID NO: 2 and SEQ ID NO: 3) using the separated DNA as a template As a result, the 16S-rRNA gene of about 1500 bp fragment was amplified in each strain.

518F primer: 5'-CCAGCAGCCGCGGTAATACG-3 '(SEQ ID NO: 1)

800R primer: 5'-TACCAGGGTATCTAATCC-3 '(SEQ ID NO: 2)

The 16S-rRNA partial sequence of the SYR1 strain determined by analysis of the PCR product of amplified DNA with 16S-rRNA sequencing analysis (Macrogen, Korea) and analyzed by Macrogen Co., Ltd. is shown in SEQ ID NO:

The results were analyzed using GenBank's Advanced BLAST (http://www.ncbi.nlm.nihm.nih.gov) using the BioEdit Sequence Alignment Editor (ver. 5.0.9, Hall, T., Nucleic Acids Symposium Series , 41: 95, gov) (Altschul et al ., Nucleic Acids Res. , 25: 3389, 1997).

As a result, it was identified as a new strain showing 97% homology with Bcillus alcalophilus (Table 1) and deposited with Accession No. KACC 91705P on January 20, 2012 to the National Institute of Agricultural Science and Technology, National Institute of Agricultural Science and Technology.

microbe 16S-rDNA
size GenBank
Accession No. Strain name Homology SYR1 1560 bp EU231621.1 Bcillus alcalophilus 97%

Characteristics of strain SYR1 microbe Colony
color shape Width Length
(탆)  Spore Gram stain motility Catalase Spore position Spore morphology SYR1 Orange-pink Short bacillus 0.5 to 1.2
2 to 4 Close to the center
Or end Oval + + +

Example 2. Identification of the protease produced by the strain SYR1

In order to confirm that the strain SYR1 isolated in Example 1 was able to dissolve parasites and kelp, 10 μl of the culture broth was applied to a parcel or tuna powder solid medium and dropped into a parcel and kelp powder solid plate culture medium, followed by drying for 4 days And cultured at 37 ° C. After colonies were identified on the culture medium formed by each strain, 10 ml of Lugol's solution (KI 10 g / L, I 5 g / L) was added to parasitic and tangle powder medium and stained for 5 minutes. , And then a clear zone was confirmed by parsley and seaweed decomposition.

As a result, it was confirmed that the SYR1 strain exhibited high resolution in the blue and kelp powder solid plate medium (FIGS. 2 and 4).

Based on the above results, in order to confirm the resolving power of the various seaweed polysaccharides (cellulose and laminarin, alginic acid, fucoidan) of the SYR1 strain, 10 μl of the culture medium of the strain was dropped into a cellulose solid plate medium and a laminarin solid plate medium, Dried, and then cultured at 37 ° C for 4 days. After colonies were identified on the culture medium formed by each strain, 10 ml of rugol solution was added to a flat culture medium of seaweed polysaccharide (cellulose and laminarin, alginic acid, fucoidan), and the mixture was dyed for 5 minutes. Then, the solution was discarded and then the solution was subjected to cellulosic and laminarin degradation And a clear zone due to the presence of the polymer.

As a result, it was confirmed that the SYR1 strain exhibits high resolution against cellulose and laminarin (FIGS. 3 and 5).

Example 3. Degradation of SYR1 in Paella and Kelp Powder

In this Example, Parasites and kelp were disassembled using the SYR1 strain to confirm the possibility of producing useful substances.

The strain SYR1 to be used in the Examples was inoculated in a 1% powdered medium for 1 minute, cultured at 30 ° C and 150 rpm for 1 to 2 days, and then used at the end of the logarithmic growth phase.

The culture was carried out by inoculating 5% of the SYR1 strain in 50 ml liquid medium (10 g / L of powdered powder, 1 g / L of NH ₄ Cl, pH 7.8) containing 1% And the decomposition reaction for kelp (kelp powder) was carried out in the same manner.

During the biodegradation reaction, samples in the flask were collected at intervals of 12 hours at the beginning, and the characteristics of the decomposition reaction were analyzed. The change in pH in the culture medium during the degradation reaction was measured using a pH meter (Istek, Korea). The concentration of the microorganism was measured at 400 nm using a spectrophotometer after threefold dilution of the sample in the cuvette. The Total Reducing Sugars concentration in the degradation reaction was determined by homogeneously mixing 100 μl of the supernatant (7000 rpm, 10 min) and 1 ml of DNS (3.5-dinitrosalicylic acid) in the culture medium, Minute, allowed to cool for 5 minutes in cold water, and measured at 570 nm using a spectrophotometer.

As a result, 10 g / L The pH values were 7.94 and 7.98 at the initial stage of culture at 10 and 10 g / L of tidal powder, respectively. The pH values were decreased to 8.29 and 8.07 at 8 days after the final culture. The density of microorganisms gradually increased and reached the highest at 1 day and 6 days And thereafter decreased at a constant concentration. The total reducing sugars produced as a result of the decomposition were 1.8 g / L (Figure 7A) and 2.0 g / L (Fig. 7B).

The total amount of reducing sugar 1.8 g / L (Fig. 6), they produced almost the same amount of reducing sugar.

Example 4. Degradation of Cellulose and Laminarin of SYR1 Strain

In this Example, the possibility that the strain SYR1 could decompose cellulose and laminarin to produce a useful substance was confirmed.

The strain SYR1 to be used for the experiment was cultured in 1% Kim powder solid medium at 30 ° C for 1-2 days and then used at the end of the logarithmic growth phase.

Each polysaccharide liquid medium, 50 ml of carboxymethylcellulose liquid medium (Carboxymethylcellulose sodium salt 1.0 g / L, MgSO ₄ .7H ₂ O 1.0 g / L, Peptone 1.0 g / L, Yeast extract 1.0 g / L, KH ₂ PO ₄ 1.0 g / L pH 7.8) and 50 ml laminarin liquid medium (Laminarin 1.0 g / L, MgSO ₄ .7H ₂ O 1.0 g / L, Peptone 1.0 g / L, Yeast extract 1.0 g / L, KH ₂ PO ₄ 1.0 g / L, pH 7.8) and cultured at 30 ° C and 150 rpm for 2 days. Experiments were carried out using the same assay method as in Example 3.

As a result, in the case of the carboxymethyl cellulose decomposition experiment, the pH value which was 7.95 at the initial stage of culturing at 1 g / L of carboxymethyl cellulose showed a slight decrease and increased to 8.16 after 2 days of the final culture. After the increase, it was increased at a constant concentration and gradually increased, but it was the highest at 36 hours. The total reducing sugar produced as a result of the digestion was 0.29 g / L (FIG. 9A).

For laminarin degradation experiments, 1 g / L of laminarin The pH value was decreased from 7.84 at the initial stage to 8.01 at 2 days after the final culture. The density of the microorganisms increased greatly after 12 hours of incubation, When the highest value was shown. The total reducing sugar produced as a result of the digestion was 0.31 g / L (FIG. 9B).

The higher reducing sugars were produced when compared to the respective total reducing sugars, 0.22 g / L (FIG. 8A) and 0.25 g / L (FIG. 8B), which were obtained from agar and carrageenan degradation experiments.

Example 5. Identification of optimal conditions for the degradation of cellulose and laminarin of strain SYR1

In addition to agar and carrageenan, SYR1 strains were found to be capable of degrading cellulose and laminarin, representative polysaccharides of green algae and brown algae.

In this example, it was confirmed that the isolated strain SYR1 was capable of degrading cellulose and laminarin, and observed under the optimal conditions over time.

Carboxymethylcellulose sodium salt (10 g / L of carboxymethylcellulose sodium salt, 1.0 g / L of MgSO ₄ .7H ₂ O, Peptone 1.0 g / L, Yeast extract 1.0 g / L, KH ₂ PO ₄ 1.0 g / L pH 7.8) and laminarin (Laminarin 10 g / L, MgSO ₄ .7H ₂ O 1.0 g / L, Peptone 1.0 g / L, Yeast extract 1.0 g / KH ₂ PO ₄ 1.0 g / L, pH 7.8) was added dropwise to the center of a solid plate culture medium supplemented with 1.5% agar for 10 days in a culture medium of SYR1 strain cultured in each polysaccharide liquid medium (0.1%) for 1 day After drying, the cells were cultured in a 30 ° C incubator.

After the SYR1 strain formed colonies on the medium, 10 ml of Lugol's solution (KI 10 g / L, I 5 g / L) was added to the carboxymethylcellulose and laminarin solid plate medium and stained for 5 minutes After discarding the solution, changes in the size of the rings with or without the formation of orange circles around the colonies on the solid plate medium stained with purple dye due to the Rugol solution were observed (Saraswathi, S. et al ., African Journal of Microbiology Research . 5: 2960, 2011).

As a result of the above experiment, it was found that around 2, 4, and 6 days after colonies cultured on a carboxymethylcellulose flat plate medium, orange rings having a radius of 1 ㎝, 1.3 ㎝ and 1.6 ㎝ were formed, It was confirmed that the size of the ring gradually increased due to the decomposition of cellulose of the SYR1 strain (Fig. 10A), and in the laminarin plate medium, the orange circles around the colonies grew longer than 2 days, 4 days, and 6 days, It was confirmed by decomposition that the radius of the ring gradually increased to 1.1 cm, 1.55 cm, and 2.03 cm (Fig. 10B).

The SYR1 isolates were found to be capable of decomposing cellulose and green alga laminarin in addition to polysaccharides, agar, and carrageenan contained in red algae under optimal conditions.

Example 6. Confirmation of Protein Resolution of SYR1 Strain

In this Example, it was confirmed whether the SYR1 strain had proteolytic ability.

10 μl of the SYR1 strain culture was dropped into the center of a solid plate culture medium supplemented with 1.5% agar in a skim milk medium (10 g / L of skim milk), followed by drying or streaking. Lt; 0 > C).

As a result of the above-mentioned method, it was confirmed that the SYR1 strain formed colonies on the medium and then cleaved the protein to form a clear zone of 2.3 cm in size (FIG. 11).

As a result, the SYR1 strain synthesizing the complex enzyme has the ability to decompose not only the polysaccharides of seaweeds and seaweeds but also the proteins, so that it will be easy to treat mixed seaweed waste mixed with fish waste in the future.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Agricultural Biotechnology Research Institute KACC91705 20120120

<110> Pukyong National University Industry-University cooperation Foundation <120> Method for Treating Mixed Seaweed Wastes Using Bacillus          alcalophilus <130> P13-B166 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 1 ccagcagccg cggtaatacg 20 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 taccagggta tctaatcc 18 <210> 3 <211> 1559 <212> DNA <213> Bcillus alcalophilus <400> 3 attttttggc gtttttggtt ttggttccct gcttcaggac gaacgctggc cggcgtgcct 60 taatccatgc aaagtcgaag cggattgatg ggaagctggt tccctgttat caccgccgaa 120 cggttgatta ccacgtggtt accttcctgg taggactggg ataattccgg gaacccgggg 180 ttattcccgg ttaattcatt tcttgccagg aggaaatgtg aaaaggtggt ttcggctatc 240 acttacagat ggacccgcgg cgcattagct agttggtgag gtaacggctc accaaggcga 300 cgatgcgtag ccgacctgag agggtgatcg gccacactgg gactgagaca cggcccagac 360 tcctacggga ggcagcagta gggaatcttc cgcaatggac gaaagtctga cggagcaacg 420 ccgcgtgagt gatgaaggtt ttcggatcgt aaagctctgt tgttagggaa gaacaagtac 480 cgttcgaata gggcggtacc ttgacggtac ctaaccagaa agccacggct aactacgtgc 540 cagcagccgc ggtaatacgt aggtggcaag cgttgtccgg aattattggg cgtaaagcgc 600 gcgcaggtgg tttcttaagt ctgatgtgaa agcccacggc tcaaccgtgg agggtcattg 660 gaaactgggg aacttgagtg cagaagagga aagtggaatt ccaagtgtag cggtgaaatg 720 cgtagatatt tggaggaaca ccagtggcga aggcgacttt ctggtctgta actgacactg 780 agcggaaaga gtggggagca aacaggatta gataccctgg tagtccacgc cgtaaacgat 840 gagtgctaag tgttaggggg tttccgcccc ttagtgctgc agctaacgca ttaagcactc 900 cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg cccgcacaag 960 cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt cttgacatcc 1020 tctgacaacc ctagagatag ggctttcccc ttcgggggac agagtgacag gtggtgcatg 1080 gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg 1140 atcttagttg ccagcattca gttgggcact ctaagatgac tgccggtgac aaaccggagg 1200 aaggtggggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca cacgtgctac 1260 aatggacggt acaaagggca gcgagaccgc gaggtttagc caatcccata aaaccgttct 1320 cagttcggat tgtaggctgc aactcgccta catgaagctg gaatcgctag taatcgcgga 1380 tcaccatgcc gcggttgaat acgttcccgg gccttgtaca caccgcccgt cacaccacga 1440 agagtttgta acacccgaag tccggtgagg taaccttttt ggagccagcc gcctaaggtg 1500 ggacagatga ttgggggtga tctacaaagg gggaggcgcg atatatataa ggagacaga 1559

Claims (5)

A method for producing a reducing sugar by decomposing a seaweed or seaweed-derived polysaccharide characterized by using Bacillus alcalophila KACC91705P.
The method according to claim 1, wherein the seaweed is selected from the group consisting of red algae, green algae and brown algae.
The method according to claim 1, wherein the seaweed-derived polysaccharide is selected from the group consisting of agar, carrageenan, laminarin, alginic acid, fucoidan and cellulose.
Characterized in that Bacillus alcalophilus KACC91705P is used.
5. The method of claim 4, wherein the seaweed waste is a seaweed waste, a fish waste or a mixed waste of seaweed waste and fish waste.

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