KR101920557B1 - Mixed Strain for Decomposing Food Waste and Method for Treating Food Waste Using the Same - Google Patents

Abstract

Bacterial strains for the treatment of food waste are Bacillus amyloliquefaciens, Acinetobacter baumannii, Enterobacter hormaechei, Saccharomycopsis fibuligera, Raoultella ornithinolytica and Serratia marcescens strains which have amylase, cellulase, protease and lipase activity and have excellent food waste decomposition activity and growth temperature range 2 to 50 캜, and a salt concentration of 3%.

Description

Technical Field [0001] The present invention relates to a mixed strain for the disposal of food waste and a method for treating the same using food waste disposal method.

The present invention relates to a food waste disposal technique, and more particularly, to a mixed waste disposal method for food waste disposal having a rapid growth rate in various temperature, pH and salinity ranges and a higher decomposition rate for cellulose, amylose, protein and fat, The present invention relates to a garbage disposal method.

Domestic food waste is very high in moisture content and organic matter content of over 80% and is high in salinity, which is easily corroded during storage, collection and transport, and acts as a deterioration factor in the residential complex due to aesthetic adverse effects such as odor and leachate .

Therefore, it is necessary to develop efficient technology for proper treatment of food waste from source, and food waste generated in apartment house is expected to be useful energy resource because it is easy to utilize after biological treatment because the amount and characteristics of food waste are almost constant have.

In addition, domestic and overseas waste treatment and resource facilities are mainly operated in a large-scale and concentrated manner with a reduction / resource-use method, but they have various technological, social and economic limitations. And the promotion of a food-weighting system. However, the effect is insignificant due to lack of recognition and practice of resource recycling.

Food waste was low in calorific value and high in water content, so it was not suitable for incineration and it relied on landfill. However, food contamination was prohibited in 2005 due to secondary pollution by odor and leachate and shortening of landfill.

In addition, since the London Convention prohibits all marine dumping of wastes from 2013, the development of new methods for treating food waste is highly required.

There is a method of disinfection using microorganisms as a method that can efficiently reduce the incidence of secondary pollution while efficiently treating food waste.

The disinfection process using these microorganisms significantly reduces the amount of food waste by fermenting the foodstuffs under aerobic conditions and suitable process in a short time and minimizes secondary environmental pollution which is pointed out as a problem of existing treatment methods. This is the most reasonable alternative.

The disinfection method using microorganisms has a treatment process of decomposing organic matter of food waste into water and carbon dioxide by volatilizing the microorganisms in a device whose biological environment is appropriately controlled and volatilizing it into the atmosphere.

(70 ~ 85%) in the food by evaporating the organic matter (about 12 ~ 13%) in the food by continuously feeding the food into the fermentation tank under proper process conditions with the aerobic condition secured. The increase in the number of users is maintained.

Currently, the microorganisms used in the food waste disposal method in Korea are bacteria such as Bacillus, Actinomycetes, Pseudomonas, Penicillium, Rhizopus, And Pichia. These microorganisms are microorganisms that have been judged to have excellent food-extinction ability and deodorizing power.

However, the ability of these microorganisms to decompose substances is insufficient, and when they are used alone, the efficiency of treatment decreases due to the death of the inoculation strain due to environmental changes in the food waste disposal device, .

For example, actinomycetes are capable of decomposing carbohydrates such as carbohydrates and fibrils and producing compounds such as geosmin that give off the soil odor (Parker, 2001) On the other hand, it is disadvantageous that the inoculated microorganisms are easily lost due to the slow propagation speed and the decomposition ability of the protein component is relatively low, which causes the ammonia and hydrogen sulfide odor.

Therefore, when using mixed strains containing various microorganisms, the disadvantages of such a microorganism can be complemented and the efficiency of food waste treatment can be improved.

In order to solve the problem of the use of a single microorganism for the treatment of food waste, Korean Patent No. 10-1161670 proposes a mixed bacterial strain for food waste disposal and a food waste disposal method using the same. However, these mixed strains also contain only two species of Bacillus bacteria, and there is a problem that the microbial growth and activity are insufficient at a high temperature, a low salinity, and a low pH, which are environment of food waste treatment.

Korean Registered Patent No. 10-1161670 (registered on June 26, 2012), entitled " Mixed strains for the treatment of food waste and method for treating food waste using the same,

In order to solve such problems, the present invention provides a mixed strain for food waste disassembly which has a rapid growth rate in various temperature, pH and salinity ranges and has a higher decomposition rate with respect to cellulose, amylose, protein and fat, and a food waste disposal method using the same The purpose is to provide.

According to an aspect of the present invention, there is provided a mixed bacterial strain for digesting food waste,

Bacillus amyloliquefaciens (Accession No. KCTC18541P), Acinetobacter baumannii (Accession No. KCTC18542P), Enterobacter hormaechei (Accession No. KCTC18543P), Saccharomyces cerevisiae A strain of Saccharomycopsis fibuligera (Accession No. KCTC18544P), Raoultella ornithinolytica (Accession No. KCTC18545P) and Serratia marcescens (Accession No. KCTC18546P), amylase, cellulase , Protease and lipase enzyme activity and is excellent in the activity of decomposing garbage, has a growth temperature range of 30 ° C, and is capable of growing to a salt concentration of 5%.

A method of treating food waste according to aspects of the present invention includes:

Bacillus amyloliquefaciens (Accession No. KCTC18541P), Acinetobacter baumannii (Accession No. KCTC18542P), Enterobacter hormaechei (Accession No. KCTC18543P), Saccharomyces cerevisiae Using a mixed strain of Saccharomycopsis fibuligera (Accession No. KCTC18544P), Raoultella ornithinolytica (Accession No. KCTC18545P) and Serratia marcescens (Accession No. KCTC18546P) .

According to the above-described constitution, the present invention has a very wide range of temperature and pH that can be grown, and has high productivity and activity of enzymes that decompose both carbohydrates, proteins, fibrin, and lipids, so that the degree of extinction of food waste is extremely high.

The present invention relates to a food and beverage product which is excellent in decomposing power for both the soluble portion and the non-soluble portion of the food, is capable of growing even under a high salt concentration, is excellent in the ability to inhibit odors generated from foods and has excellent heat resistance, acid resistance, There is a very useful effect in destroying.

1 is a view showing a culture state in the evaluation of salt tolerance of a mixed strain according to an embodiment of the present invention.
2 is a view showing a culture state in the heat resistance evaluation of a mixed strain according to an embodiment of the present invention.

Throughout the specification, when an element is referred to as " comprising ", it means that it can include other elements as well, without excluding other elements unless specifically stated otherwise.

(1) Isolation and selection of microorganisms

In order to isolate a strain capable of decomposing food waste which is excellent in salt resistance and can live at high temperature, samples are taken from food waste and koji which are generated in ordinary households and shopping malls.

10 g of each sample is immersed in sterilized distilled water for 3 hours and then cultured on a nutrient agar (0.3% beef extract, 0.5% peptone, 1.5% agar) medium and cultured at 30 ° C. for 24 hours.

After 24 hours, different colonies are selected according to the size and shape of colony on the medium, and transferred to nutrient agar medium to isolate the strain.

As shown in Table 1 and Table 2, in order to isolate the strains, food waste generated in one household (one room, one living room, etc.) Modified nuruk, pearl crab, Songjak, and acidic yeast). 45 strains from food waste and 72 strains from yeast were isolated and cultured.

(2) Identification of microorganisms

Bacillus amyloliquefaciens (99% identical), Acinetobacter baumannii (99% identical), Enterobacter hormaechei (99% identical) and Saccharomycopsis fibuligera (99% identical) were compared with NCBI BLAST 99% identical), Raoultella ornithinolytica (99% identical) and Serratia marcescens (99% identical).

The present inventors deposited a strain of Bacillus amyloliquefaciens of the present invention on February 6, 2017 at the KCTC (International Collection Service for Type Cultures) BRC under the Budapest Treaty, and deposited the deposit number KCTC18541P .

The present inventors deposited the strain of Acinetobacter baumannii of the present invention on February 6, 2017 in the KCTC (International Collection Service for Type Cultures) BRC under the Budapest Treaty, and deposited the deposit number KCTC18542P .

The present inventors deposited a strain of Enterobacter hormaechei of the present invention on February 6, 2017 in the KCTC (International Collection Service for Type Cultures) BRC under the Budapest Treaty, and deposited the deposit number KCTC18543P .

The inventors of the present invention deposited a strain of Saccharomycopsis fibuligera of the present invention on February 6, 2017 at the KCTC (International Collection Service for Type Cultures) BRC under the Budapest Treaty, No. KCTC18544P.

The present inventors deposited a strain of Raoultella ornithinolytica of the present invention on February 6, 2017 at the KCTC (International Collection Service for Type Cultures) biological resource center under the Budapest Treaty, KCTC18545P.

The present inventors deposited a strain of Serratia marcescens of the present invention on February 6, 2017 at the KCTC (International Collection Service for Type Cultures) BRC under the Budapest Treaty, and deposited the deposit number KCTC18546P received.

The mixed strain of the present invention has amylase, cellulase, protease and lipase enzyme activity, is excellent in the activity of decomposing garbage, is a strain for treating food waste which can grow up to a growth temperature of 30 ° C and a salt concentration of 5%.

The above-mentioned mixed strains can be applied at a survival temperature of 10 ° C to 50 ° C, a salt content of 25% or less, and a pH range of 3.0 to 11.0.

The salt tolerance, heat resistance and enzyme activity of the mixed strains according to the examples of the present invention are evaluated.

(3) Evaluation of salt tolerance of mixed strains

In order to measure the salt tolerance of the mixed strains, the growth of the strains in the medium with different salt concentration was confirmed. NaCl was added at 0%, 3%, and 5% concentrations in the preparation of Nutrient agar medium. 10 μl of the culture broth cultured in the liquid medium was inoculated in a medium having different salt concentration and cultured at 30 ° C. for 24 hours. Determine the salt tolerance based on the size and clarity of the colony.

As shown in FIG. 1, [Table 3] and [Table 4], the growth of mixed strains was confirmed at 0%, 3% and 5% salinity by the above experimental method. If the salt concentration in the Korean food is about 3% or less, it can be used as a food waste disintegrating strain.

Of the 74 strains isolated from Nuruk, 63 strains were able to grow at 3% salinity and 58 strains at 5% salinity. Of the 45 strains isolated from food waste, 41 strains can be cultured at 5% salinity in all strains at 3% salinity.

(3) Evaluation of heat resistance of mixed strains

In order to measure the heat resistance of the isolated strain, the growth of the strain at 30 ° C, 40 ° C and 50 ° C is confirmed. 10 μl of the culture medium previously cultured in the liquid medium is inoculated into a nutrient agar medium and cultured at each temperature for 24 hours. Check the heat resistance based on the size and clarity of the colony.

As shown in FIG. 2, [Table 5] and [Table 6], the possibility of the growth of the isolated strain at the temperatures of 30 ° C, 40 ° C and 50 ° C was confirmed by the above experimental method. As a result of culturing the isolated strain at each temperature, it is possible to cultivate 58 strains at 40 ° C and 32 strains at 50 ° C among 74 strains isolated from the yeast. Among 45 strains isolated from food waste, 40 strains can be cultured at 40 ° C and 2 strains can be cultured at 50 ° C.

(4) Evaluation of enzyme activity of mixed strains

Amylase, cellulase, lipase, and protease activities are investigated in order to measure the degradation ability of isolated strains.

The present invention uses a paper disc method to select a strain having a high enzyme activity, and a solid sorting medium containing a substrate component that specifically reacts with each enzyme is used.

Amylase activity was determined by 8 mm paper discs on starch agar (0.3% beef extract, 1% soluble starch, 1.2% agar) and 20 μl of each culture was dispensed at 30 ° C for 24 hours. And the resolution is checked to the diameter of the clear zone.

For the confirmation of cellulase activity, 8 mm paper discs were placed on a nutrient agar medium containing 1% CMC, and 20 μl of each culture was dispensed. The cells were cultured at 30 ° C for 24 hours, stained with 0.1% congo red for 30 minutes, After washing, examine the diameter of the inhibition ring.

Lipase activity was measured by placing 8 mm paper discs on sierra lipolytic agar (1% peptone, 0.5% sodium chloride, 0.5% calcium chloride, 1.5% agar) medium supplemented with 1% Lt; 0 > C for 24 hours to examine the resolution of the diameter of the inhibition ring.

Protease activity was measured by placing 8-mm paper discs on nutrient agar medium supplemented with 1% skim milk, and dividing 20 μl of each culture medium by 30 ° C for 24 hours.

The size of the inhibitory ring around the disc after culturing was measured. +, 20 ~ 30 mm ++, and 30 mm or more were used for the detection of strains with enzyme activity among isolated strains. +++.

Four isolates with amylase activity, one isolate with cellulase activity, eight isolates with lipase activity, and 17 isolates with protease activity were obtained from 72 strains isolated from Nuruk.

As shown in Table 7 and Table 8, among 45 strains isolated from food waste, three strains showing amylase activity, two strains showing cellulase activity, three strains showing lipase activity , and 8 isolates showing protease activity.

The strains JIF16003, JIF16016, JIF16026, JIF16040, JIF16045, JIF16209, JIF16238 and JIF16241 having two or more enzyme activities were selected.

(5) Re-evaluation of enzyme activity of the selected strain

The strains JIF16003, JIF16016, JIF16026, JIF16040, JIF16045, JIF16209, JIF16238 and JIF16241, which have two or more enzymatic activities among the strains isolated from yeast and food waste, were selected.

8 strains were subcultured in 5 ml nutrient broth and inoculated into 500 ml nutrient broth and cultured at 30 ° C for 24 hours. The enzyme activity was examined by the same method as above using this culture solution. The size of the inhibitory ring differs from that of the previous experiment, but the activity of the enzyme does not change after mass culture.

The names of the identified strains in the above-mentioned [Table 9] are as follows.

JIF16003 is Bacillus amyloliquefaciens, JIF16016 is Acinetobacter baumannii, JIF16026 is Enterobacter hormaechei, JIF16040 is Saccharomycopsis fibuligera, JIF16045 is Enterobacter hormaechei, JIF16209 is Raoultella ornithinolytica, JIF16238 is Serratia marcescens and JIF16241 is Raoultella ornithinolytica.

As a result of identification, strains such as JIF16026 and JIF16045, and JIF16209 and JIF16241 are the same strains.

Bacillus amyloliquefaciens is a class of bacteria belonging to the genus Bacillus, the source of the limiting factor BamH1, which synthesizes protein barnase, an antibiotic substance.

Protein barnase is a widely studied RNA hydrolase, characterized by its ability to form complexes with barstar, an inhibitor of it, and plantazolicin, an antibiotic selective to Bacillus anthracis.

Acinetobacter baumannii is becoming increasingly important as an opportunistic infectious disease that infects people with weakened immune systems and a hospital-acquired infection.

Species are Enterobacter hormaechei, Family is Enterobacteriaceae, Genus is Enterobacter.

Enterobacter is a member of the Enterobacteriaceae family, usually gram-negative, coarse anaerobic, rod-shaped, nonporosity-forming bacteria. Some strains of this strain are infected by pathogenic and immunocompromised hosts.

Species are Raoultella ornithinolytica, Family is Enterobacteriaceae, Genus is Raoultella.

Raoultella has Gram negative, oxidase negative, aerobic, non-motile, and flowable anaerobic characteristics.

Species are called Serratia marcescens, and Genus is called Serratia.

Serratia is Enterobacteriaceae and gram-negative, flowable anaerobic, rod-shaped bacteria. The most common species of this species, S. marcescens, is generally pathogenic and causes hospital infections. However, the rare strains S. pneumatica, S. liquefaciens, S. rubidaea, and S. odoriferae have caused disease through infection. Members of this genus produce reddish pigment and produce prodigiosin characteristically. And can be distinguished from other members of the Enterobacteriaceae by their specificity for the production of their three enzymes (DNase, lipase, and gelatinase).

Institution name: Korea Biotechnology Research Institute

Access number: KCTC18541P

Checked on: 20170206

Institution name: Korea Biotechnology Research Institute

Access number: KCTC18542P

Checked on: 20170206

Institution name: Korea Biotechnology Research Institute

Accession number: KCTC18543P

Checked on: 20170206

Institution name: Korea Biotechnology Research Institute

Access number: KCTC18544P

Checked on: 20170206

Institution name: Korea Biotechnology Research Institute

Accession number: KCTC18545P

Checked on: 20170206

Institution name: Korea Biotechnology Research Institute

Access number: KCTC18546P

Checked on: 20170206

The embodiments of the present invention described above are not implemented only by the apparatus and / or method, but may be implemented through a program for realizing functions corresponding to the configuration of the embodiment of the present invention, a recording medium on which the program is recorded And such an embodiment can be easily implemented by those skilled in the art from the description of the embodiments described above.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, It belongs to the scope of right.

Claims (3)

Bacillus amyloliquefaciens (Accession No. KCTC18541P), Acinetobacter baumannii (Accession No. KCTC18542P), Enterobacter hormaechei (Accession No. KCTC18543P), Saccharomyces cerevisiae A strain of Saccharomycopsis fibuligera (Accession No. KCTC18544P), Raoultella ornithinolytica (Accession No. KCTC18545P) and Serratia marcescens (Accession No. KCTC18546P), amylase, cellulase , A protease and a lipase enzyme activity, is excellent in the activity of decomposing garbage, has a growth temperature range of 30 ° C, and can grow up to a salt concentration of 5%. delete Bacillus amyloliquefaciens (Accession No. KCTC18541P), Acinetobacter baumannii (Accession No. KCTC18542P), Enterobacter hormaechei (Accession No. KCTC18543P), Saccharomyces cerevisiae A mixed strain of Saccharomycopsis fibuligera (Accession No. KCTC18544P), Raoultella ornithinolytica (Accession No. KCTC18545P) and Serratia marcescens (Accession No. KCTC18546P) How to treat food waste.

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