JP2006501810A - Pseudomonas genus FK916 producing organic matter degrading enzyme and method for treating organic waste using the same - Google Patents

Abstract

The present invention relates to a Pseudomonas genus FK916 strain that produces an organic matter-degrading enzyme and a method for treating organic waste using the same. This Pseudomonas genus FK916 strain is isolated from sewage, has excellent ability to decompose and dissipate organic matter, and decomposes at a temperature of 15 to 35 ° C., so it produces almost no bad odor and is generated from food waste and factories. It has an excellent effect of treating organic waste.

Description

  The present invention relates to a Pseudomonas genus FK916 (Pseudomonas sp. FK 916) that produces an enzyme that degrades organic compounds, and a method for treating organic waste using the same. More particularly, the present invention is excellent in the ability to decompose and eliminate food waste and organic matter in organic waste separated from sewage and effectively treat food waste without the emission of malodors. The present invention relates to a Pseudomonas genus FK916 strain that produces an organic matter-degrading enzyme and a method for treating organic waste using the same.

  The amount of food waste generated in South Korea is 11,577 tons / day (based on 1999), accounting for approximately 26.5% of the amount of solid waste generated in the city. % Come from restaurants and restaurants. Since 58.8% of the generated food waste is landfilled, 7.3% is incinerated, and 33.9% is treated by recycling, the situation depends on landfill. In the case of the Tokyo metropolitan area, direct landfill of food waste in the Tokyo metropolitan area landfill has been prohibited since January 2005, and most local self-governing bodies have also caused environmental pollution caused by leachate in the landfill area and the surrounding area of the landfill area. Food waste disposal problems are becoming more and more serious as the grudges living in the country increase. Also, incineration treatment has a high moisture content, so it costs more to treat than combustible waste. Construction of an incineration treatment plant and securing of the site are very difficult due to the opposition of residents. Therefore, in order to solve such problems, economic processing that can decompose, eliminate, or reduce food waste directly with microorganisms at the place where food waste is generated, or increase the reuse rate. The method needs to be further developed.

  Food waste is not uniform in nature and has a high moisture and salt content, and it is very difficult to dispose of it. It is necessary to search for various treatment methods by region and business. Technologies considered as a method for the treatment and reuse of food waste include composting, feed conversion, anaerobic digestion, and combined sewage treatment. It can be divided into two main categories: small composting technology that can be used in stores.

  Small composting equipment consists of food waste fermenters and dryers and is installed in apartment complexes, group lunchhouses, hotels, restaurants, etc., but it has various problems.

  The problem with conventional technology and devices that ferment food waste at high temperature and reuse it as compost and feed is that, firstly, because fermentation or rot progresses at high temperature, a severe odor is generated and it is avoided as an aversion facility In addition, since Korean foods usually contain 1 to 5% high-concentration salt, it is not suitable for use as a fertilizer. Second, to increase the rate of fermentation degradation by microorganisms, use thermophilic bacteria and actinomycetes to maintain the optimal growth temperature for the growth of high-temperature microorganisms, to reduce the volume of food waste fermentation products, and to evaporate water Since the temperature inside the apparatus must be maintained at 50 to 60 ° C., there is a disadvantage that a large amount of electricity is consumed and the operation cost is extremely increased.

  In order to solve the above-mentioned problems, the present inventor is excellent in the ability to decompose and eliminate organic matter in food waste and organic waste from sewage, and does not generate bad odor by aerobic decomposition at medium temperature. A new strain was isolated and identified, and the present invention was completed by using this strain for decomposition and elimination of food waste.

  Therefore, the objective of this invention is providing the strain excellent in the capability to decompose | disassemble and extinguish the organic substance in food waste and organic waste.

  Another object of the present invention is to provide a waste treatment method using the strain of the present invention.

  According to the present invention, these and other objects can be achieved by providing Pseudomonas FK916.

Pseudomonas genus FK916 separated from sewage removes organic substances such as carbohydrates and fats in food waste by aerobic decomposition in the presence of sufficient oxygen at a temperature of 15 to 35 ° C, preferably 25 to 30 ° C. It is converted into a stable low-molecular-weight organic substance, and as a final product, stable substances such as CO ₂ and H ₂ O are generated, and a little heat is generated. Pseudomonas microorganisms have the following reaction formula (I)

As shown in FIG. ₃ , NO ₃ , NO ₂ , NO can be denitrified to N ₂ , so that Pseudomonas FK916, together with other microorganisms, undergoes a denitrification process that converts NH ₃ which is one of malodorous gas components into N _2. In addition, the malodor and irritation of ammonia can be removed.

  In addition, the strain of the present invention has characteristics that do not produce malodorous components such as indole, hydrogen sulfide, and acetoin (Table 2).

  The strain of the present invention, Pseudomonas genus FK916, was deposited with the Korean inoculation association (KCCM), an international depositary organization, as deposit number KCCM-10350 on January 7, 2002. In addition to the deposited strains, mutant strains of the strains that have some changes in other characteristics also have the same 16S rRNA base sequence and fatty acid composition, morphology, culture, and biochemical characteristics of the present invention. As long as it is included in the scope of the present invention.

  The present invention also provides a method for decomposing and eliminating organic waste such as food waste using Pseudomonas FK916 strain.

  The method is characterized in that a Pseudomonas genus FK916 strain or a composition containing the strain is added to an object to be treated. As a method for treating the strain or a composition thereof, the culture solution of the strain is directly sprayed, or the strain culture solution is coated with natural particles such as rough slag, sawdust and wood chips, inorganic particles such as zeolite and ceramic. In addition, it can be mixed with a voiding agent such as plastics, or adsorbed on the above-mentioned natural organic matter, inorganic particles, plastics or other voiding agent or carrier and sprayed on organic waste, or processed normally. It can be formulated and used by a method for producing a microbial preparation.

  In the present invention, the “object to be treated” means not only food waste, but also activated sewage sludge, organic waste generated from a factory, and sewage waste.

  The above and other objects, features and advantages of the present invention will be more clearly understood from the following detailed description based on the accompanying drawings.

  FIG. 1 shows the results of confirming the activity of a proteolytic enzyme by inoculating Pseudomonas FK916 according to the present invention on a proteolytic enzyme search agar plate.

  FIG. 2 shows the result of inoculating Pseudomonas sp. FK916 on a lipolytic enzyme search agar plate and confirming the activity of the lipolytic enzyme.

  FIG. 3 is a scanning electron micrograph of Pseudomonas genus FK916.

  FIG. 4 is a graph showing the food waste reduction rate according to the decomposition time after adding a culture solution of Pseudomonas FK916 to food waste.

  Hereinafter, the present invention will be described in more detail based on examples. Those skilled in the art will understand that these examples are for illustrative purposes only and are not intended to limit the present invention.

[Example 1: Isolation of strains]
The strain according to the present invention was isolated from a sample collected from sewage near Jeollabuk-do, Korea. This sample was appropriately diluted stepwise with physiological saline, and a part thereof was LB agar medium (bactotryptone 10 g / L, bacto yeast extract 5 g / L, NaCl 10 g / L, agar 15 g / L, pH 7.0). And the colonies that appeared were separated.

The isolated strain was cultured and grown in a strain growth medium (LB liquid medium) at 25 ° C., followed by proteolytic enzyme search agar medium (defatted milk agar medium: peptone 5 g / L, bacto yeast extract 3 g / L, skim milk 1 g / L, agar 15 g / L, pH 7.0), lipolytic enzyme search agar medium (tributyrin agar: peptone 5 g / L, bacto yeast extract 3 g / L, tributyrin 5 mL / L, agar 15 g / L, pH 7 .4), starch-degrading enzyme search agar medium (starch agar medium: peptone 5 g / L, bacto yeast extract 3 g / L, soluble starch 2 g / L, agar 15 g / L, pH 7.0), cellulose search agar medium (cellulose 1 g / L, K ₂ HPO ₄ 1 g / L, (NH ₄ ) ₂ SO ₄ 1 g / L, MgSO ₄ .7H ₂ O 0.2 g / L, CaCl ₂ .2H ₂ O 0.1 g / L, FeC l ₃ 0.02 g / L, Congo Red 0.075 g / L, agar 15 g / L, pH 7.2) smeared on a plate, depending on the activity of proteolytic enzyme, lipolytic enzyme, starch degrading enzyme, cellulose degrading enzyme The presence or absence of the resulting transparent ring was confirmed and a positive strain was searched for and separated.

  As a result of the experiment, as shown in FIG. 1, when the FK916 strain was inoculated on the proteolytic enzyme search agar plate (A) and the lipolytic enzyme search agar plate (B), a transparent ring was formed. Was found to form proteolytic enzymes and lipolytic enzymes. However, the activity of amylolytic enzyme was low, and the activity of cellulolytic enzyme was not recognized.

[Example 2: Identification of strain]
The isolated strain is a strain belonging to the genus Pseudomonas, identified by a method such as Holt (Holt JG, et al., Bergey, s Manual of Determinative Bacteriology, 9th ed. (1994)), Sherlock system and clustal W. And was named Pseudomonas genus FK916. The fatty acid and 16S rRNA of Pseudomonas genus FK916 represented the fatty acid composition of Pseudomonas chlororaphis and 80.2%, and the similarity of 169.8 rRNA of Pseudomonas gessardii CIP 105469 and 99.8%, respectively.

  As can be seen from the scanning micrograph of FIG. 3, the strain was a gonococcus and a gram-negative bacterium. As shown in Table 1 below, the optimal growth temperature is 28-30 ° C. and the culture characteristics are 6% resistant to NaCl. Therefore, the strain of the present invention treats high-salt food waste. It turns out that it is suitable to do.

  The biochemical characteristics analyzed by the API kit are shown in Table 2, and the fatty acid analysis of whole cells analyzed by gas chromatography by the method presented by MIDI and the strain identification results by the Sherlock system are shown in Table 3. In addition, as a result of analyzing 16S rRNA and analyzing its base sequence by the method of Verhille et al. (Verhille, S., et al., International J. Systematic Bacteriol., 49, 1559, 1999), 1498 bases (sequence 1) The results of comparing the homology between base sequences analyzed by clustal W for the similarity of 16s rRNA base sequences between Pseudomonas sp. This is shown in FIG. The G + C content analyzed by the method of De Ley (De Ley, J., J. Bacteriol., 101, 738, 1970) was 61.5%.

[Example 3: Decomposition and extinction test of food waste: Reduction rate of food waste due to decomposition time]
In order to test the reduction of food waste, Pseudomonas sp. FK916 was inoculated into a sterilized LB liquid medium and cultured with shaking at 38 ° C. overnight. 100 g of this culture solution and sawdust (aeration and moisture regulator) are added to a 3 L beaker and mixed well, and 1 kg of food waste collected from the W University cafeteria (water content of about 80%) is added and mixed well. It was put into a C incubator, the change in weight with time was weighed, the reduction rate was calculated, and it was stirred well 3 to 4 times every day.

  As shown in FIG. 4, from the first day of decomposition, the odor peculiar to food waste disappeared, no new odor was generated, and weight reduction began. Reduced to 72.3% on the 18th day of decomposition, food waste was decomposed and extinguished. Compared to Comparative Example 1 in which only the culture solution was added, and Comparative Example 2 in which LB medium and sawdust were added, a faster weight reduction rate was achieved. Indicated. Such a result shows that the addition of sawdust satisfies aerobic conditions, the regulation of moisture promotes the decomposition and disappearance of organic matter, and the FK916 strain promotes the degradation and disappearance of organic matter at 25 ° C. On the other hand, such a result shows that a better result is obtained in food waste to which air is continuously supplied by stirring.

[Comparative Example 1: Decomposition rate measurement when food waste is treated with Pseudomonas FK916 culture solution]
A sterilized LB liquid medium was inoculated with Pseudomonas sp. FK916, and cultured with shaking at 28 ° C. overnight. Add 200 mL of this culture solution and 1 Kg of collected food waste (water content approximately 80%) to a 3 L beaker, mix well, place in a 25 ° C. incubator, weigh the change in weight over time, and calculate the reduction rate. Stir well 3-4 times daily (FIG. 4).

[Comparative Example 2: Decomposition rate measurement when food waste is treated with sawdust]
Add 200 mL of LB medium sterilized in a 3 L beaker and 100 g of sawdust (moisture control agent) and mix well, add 1 kg of collected food waste (water content approximately 80%) and mix well, then incubate at 25 ° C It was put into a vessel, the change in weight over time was weighed, the weight loss rate was calculated, and it was stirred well 3 to 4 times daily (FIG. 4).

[Example 4: Decomposition and extinction in a food waste extinguisher]
80 kg of wood chips (approximately 5 to 10 mm) in an extinguisher (CLEANGATE-50 model, capacity 50 kg) manufactured by Nambang E & B Co., Ltd. 5 L of FK916 culture solution cultured by the method of Example 1, collected at a restaurant 39 kg of food waste was added, the mixture was operated at 28 ° C. with a stirring speed of 3.2 rpm and an aeration speed of 5.5 m ³ / min, and the weight after 24 hours was weighed to calculate the reduction rate. The odor peculiar to food waste began to be removed 1 hour after the start of the decomposition, and after 24 hours the amount was reduced to 76.9%, and almost no bad odor was generated. On the second day, 46 kg of food waste was further added, the operation was performed under the same conditions, and the weight reduction rate when measured after 24 hours was 90.9%, and 27 to 60 kg of food waste was continuously applied for 29 days. When added and operated under the same conditions, the final cumulative weight loss rate was 94.7%.

  As a result of the experiment, Pseudomonas genus FK916 of the strain of the present invention decomposes and disappears food waste over a long period of time even at a single administration at normal temperature or medium temperature, and is used for the decomposition and disappearance of organic waste such as food waste. It was confirmed that this is an excellent strain.

  As explained based on the above-mentioned examples, the Pseudomonas genus FK916 strain of the present invention has an excellent ability to decompose and disappear food waste while generating no foul odor at an intermediate temperature, and can treat food waste economically and hygienically. Since it can, it is a very useful invention in the environmental industry.

It is a figure which shows the result of inoculating Pseudomonas genus FK916 by this invention on a proteolytic enzyme search agar plate, and confirming the activity of the proteolytic enzyme. It is a figure which shows the result of having inoculated Pseudomonas genus FK916 on the lipolytic enzyme search agar plate, and confirming the activity of a lipolytic enzyme. It is a figure which shows the scanning electron micrograph of Pseudomonas genus FK916. It is a graph which shows the weight loss rate of the food waste by decomposition | disassembly time, after adding the culture solution of Pseudomonas genus FK916 to food waste.

Claims (4)

  Pseudomonas sp. FK916 (deposit number KCCM 10350) which is separated from sewage and has the ability to decompose and remove food waste without generating malodor at an intermediate temperature of 15 to 35 ° C.   An organic waste and sewage treatment method comprising adding the Pseudomonas genus FK916 strain (KCCM 10350) according to claim 1 to an object to be treated.   3. The organic waste and sewage treatment method according to claim 2, wherein the Pseudomonas genus FK916 strain is added by being mixed with one selected from a pore agent such as natural organic matter, inorganic particles, and plastic.   4. The organic waste and sewage treatment method according to claim 3, wherein the natural organic matter is selected from the group of rough cocoons, sawdust, and wood chips, and the inorganic particles are selected from the group of zeolite and ceramic.

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