CN117965383A - Kitchen waste liquefaction degradation dominant bacterium combination and application thereof - Google Patents
Kitchen waste liquefaction degradation dominant bacterium combination and application thereof Download PDFInfo
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- CN117965383A CN117965383A CN202410177159.4A CN202410177159A CN117965383A CN 117965383 A CN117965383 A CN 117965383A CN 202410177159 A CN202410177159 A CN 202410177159A CN 117965383 A CN117965383 A CN 117965383A
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- sphingobacterium
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention provides a kitchen waste liquefaction degradation dominant bacterium combination and application thereof. The composition comprises the following components of non-sugar-decomposing bacillus pseudochrous, alcaligenes faecalis, bacillus amyloliquefaciens, serratia marcescens, sphingobacterium aquaticum, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingobacterium fumigatus and Sphingobacterium maltophilium. The combined bacteria can effectively realize the reduction of kitchen waste, the height can be reduced by 14.66 percent at the highest, and the COD removal rate can reach 98.19 percent at the highest. The assembled bacteria can efficiently degrade and liquefy the kitchen waste, can realize higher decrement rate and liquefaction effect, and is beneficial to the absorption and utilization of nutrient substances in the subsequent process of converting the biological resources of the kitchen waste.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a kitchen waste liquefaction degradation dominant bacterium combination and application thereof.
Background
Along with the rapid development of the catering industry, the kitchen waste production is rapidly increased, and the method has the characteristics of high yield, high pollutant concentration and the like. If not properly treated, serious pollution is caused to the environment, and the normal life of part of organisms is affected. The traditional kitchen waste treatment method has the defects of low cost and high efficiency, but microorganisms cannot directly utilize macromolecular substances in the kitchen waste. Therefore, after the kitchen waste is liquefied, the utilization rate of microorganisms on nutrient substances in the sewage can be obviously improved, and the subsequent purification and resource utilization are facilitated. The development of a method capable of rapidly degrading and liquefying kitchen waste is a key point of efficient treatment of the kitchen waste.
Disclosure of Invention
The invention aims to provide a kitchen waste liquefaction degradation dominant bacterium combination and application thereof.
In order to achieve the above object, the present invention provides the following technical solutions:
The invention provides a dominant bacteria composition, which comprises pseudomonas saccharolytica, alcaligenes faecalis, paenibacillus amyloliquefaciens, serratia marcescens, sphingobacterium aquaticum, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingobacterium fumigatus, stenotrophomonas maltophilia, sphingobacterium humicola, latin is Pseudochrobactrum asaccharolyticum、Alcaligenes faecalis、Paenibacillus amylolyticus、Serratia marcescens、Sphingobacterium mizutaii、Bacillus subtilis、Achromobacter xylosoxidans、Bacillus cereus、Sphingobacterium tabacisoli、Stenotrophomonas maltophilia、Sphingobacterium humi.
In the invention, various strains in the assembled bacteria can be obtained by conventional separation and screening means.
The invention also provides application of the assembly bacteria in degrading liquefied kitchen waste.
Preferably, the assembly bacteria are formed by mixing single bacteria enrichment liquid of 11 bacteria.
The single bacteria enrichment medium comprises the following components: beef extract 3.0g/L, peptone 10.0g/L, naCl 5.0g/L, pH 7.2-7.4.
Preferably, the initial biomass of 11 bacteria of the combined strains is 0.01-0.03 g/L, and the total initial biomass after mixing is 0.11-0.33 g/L.
Preferably, when the assembly bacteria degrade kitchen waste, the running water spraying time of the kitchen waste integrated liquefying device is 120s, and the interval time is 10min
Preferably, when the assembly bacteria degrade the kitchen waste, the bacterial liquid adding time of the integrated liquefying device for the kitchen waste is 20s, and the interval time is 3h
Preferably, when the assembly bacteria degrade kitchen waste, the waste stirring time of the kitchen waste integrated liquefying device is 5min, and the interval time is 5min
Preferably, when the assembly bacteria degrade the kitchen waste, the treatment temperature of the kitchen waste integrated liquefying device is 30-45 ℃.
Preservation description
Biological material 1
Classification naming of biological materials: pseudomonas saccharolytica (Pseudochrobactrumasaccharolyticum).
Strain number of biological material: HL-1.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27442.
Biological material 2
Classification naming of biological materials: alcaligenes faecalis (ALCALIGENESFAECALIS).
Strain number of biological material: HL-2.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27443.
Biological material 3
Classification naming of biological materials: achromobacter xylosoxidans (Achromobacterxylosoxidans).
Strain number of biological material: HL-3.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27444.
Biological material 4
Classification naming of biological materials: bacillus amyloliquefaciens (Paenibacillusamylolyticus).
Strain number of biological material: HL-4.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27445.
Biological material 5
Classification naming of biological materials: bacillus cereus.
Strain number of biological material: HL-14.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27446.
Biological material 6
Classification naming of biological materials: sphingobacterium fumigatus (Sphingobacteriumtabacisoli).
Strain number of biological material: HL-15.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27447.
Biological material 7
Classification naming of biological materials: serratia marcescens (SERRATIA MARCESCENS).
Strain number of biological material: HL-18.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27448.
Biological material 8
Classification naming of biological materials: stenotrophomonas maltophilia (Stenotrophomonasmaltophilia).
Strain number of biological material: HL-20.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27449.
Biological material 9
Classification naming of biological materials: sphingobacterium humus (Sphingobacterium humi).
Strain number of biological material: HL-25.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27450.
Biological material 10
Classification naming of biological materials: sphingobacterium aquaticum (Sphingobacteriummizutaii).
Strain number of biological material: HL-26.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27451.
Biological material 11
Classification naming of biological materials: bacillus subtilis (Bacillus subtilis).
Strain number of biological material: HL-27.
Preservation unit name of biological material: china general microbiological culture Collection center (China Committee for culture Collection).
The preservation unit of biological materials is abbreviated as: CGMCC.
Deposit unit address of biological material: north Star Xilu 1, 3 of the Chaoyang district of Beijing, the institute of microbiology, national academy of sciences, postal code: 100101.
Preservation date of biological material: 2023, 05, 23.
Accession numbers of the collection center of biological materials: CGMCC No.27452.
The invention provides a composite bacterium for degrading liquefied kitchen waste and application thereof, the composite bacterium comprises 11 bacteria, namely, pseudomonas saccharolytica, alcaligenes faecalis, paenibacillus amyloliquefaciens, serratia marcescens, sphingosine-aquarius, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingosine-fumigatus, stenotrophomonas maltophilia and sphingosine-humate, the seed retaining numbers of Latin are Pseudochrobactrum asaccharolyticum、Alcaligenesfaecalis、Paenibacillus amylolyticus、Serratia marcescens、Sphingobacterium mizutaii、Bacillus subtilis、Achromobacterxylosoxidans、Bacillus cereus、Sphingobacterium tabacisoli、Stenotrophomonas maltophilia、Sphingobacterium humi. and CGMCCNO.27442、CGMCCNO.27443、CGMCCNO.27445、CGMCCNO.27448、CGMCCNO.27451、CGMCCNO.27452、CGMCCNO.27444、CGMCCNO.27446、CGMCCNO.27447、CGMCCNO.27449、CGMCCNO.27450. respectively, the composite bacterium can still realize good continuous degradation effect on the kitchen waste in practical pilot plant application, and the waste reduction rate can reach 99.78% in one week. The culture condition of the composite bacteria is easy to realize and control, can effectively degrade and liquefy the kitchen waste, and has great significance for the efficient treatment of the kitchen waste.
Drawings
FIG. 1 shows the maximum degradation rates of protein (a), cellulose (b), lipid (c) and starch (d) by a single strain in example 2.
FIG. 2 shows the degradation rates of the complex bacteria (11) of example 2 on proteins, starches, oils and cellulose.
FIG. 3 shows the changes of the components of the mixed solution of kitchen waste treated with the composite bacteria in example 3 for 6 days.
FIG. 4 shows the change in the refuse elevation before and after refuse feeding in the pilot plant of example 4 (the abscissa 1-6 in the figure shows different sampling points for collecting water. 1 before refuse feeding in breakfast, 2 after refuse feeding in breakfast, 3 before refuse feeding in lunch, 4 after refuse feeding in lunch, 5 before refuse feeding in dinner, 6 after refuse feeding in dinner)
FIG. 5 shows the reduction rate of the kitchen waste height after the kitchen waste is degraded by the composite bacteria for one night (14 h) in the embodiment 4 (the abscissa of the figure is 1-6, respectively, of the waste height versus time).
FIG. 6 shows the COD removal rate of the kitchen waste liquefaction effluent after the composite bacteria in example 4 degrade overnight (14 h) (the abscissa of the graph 1-6 is the COD removal rate versus time, respectively).
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The invention provides a kitchen waste liquefaction degradation dominant bacterium composition and application thereof, wherein the composition bacterium comprises pseudomonas saccharolytica, alcaligenes faecalis, paenibacillus amyloliquefaciens, serratia marcescens, sphingobacterium aquaticum, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingobacterium fumigatus, stenotrophomonas maltophilia and Sphingobacterium humicola, and Latin is Pseudochrobactrum asaccharolyticum、Alcaligenes faecalis、Paenibacillus amylolyticus、Serratia marcescens、Sphingobacterium mizutaii、Bacillus subtilis、Achromobacter xylosoxidans、Bacillus cereus、Sphingobacterium tabacisoli、Stenotrophomonas maltophilia、Sphingobacterium humi.
The invention also provides application of the assembly bacteria in degrading liquefied kitchen waste.
In the invention, the assembly bacteria are inoculated into the kitchen waste liquefying integrated treatment device.
In the present invention, the initial biomass of 11 bacteria in the group of bacteria is the same, preferably 0.01 to 0.03g/L, more preferably 0.02g/L, and the total initial biomass is 0.11 to 0.33g/L, more preferably 0.22g/L.
In the invention, when the assembly bacteria degrade the kitchen waste, the running water spraying time of the kitchen waste integrated liquefying device is preferably 110-130 s, more preferably 120s, and the interval time is preferably 5-15 min, more preferably 10min.
Preferably, when the assembly bacteria degrade the kitchen waste, the bacterial liquid adding time of the integrated liquefying device for the kitchen waste is preferably 10-30 s, more preferably 20s, and the interval time is preferably 2-3 h, more preferably 3h.
Preferably, when the assembly bacteria degrade the kitchen waste, the waste stirring time of the kitchen waste integrated liquefying device is preferably 4-6 min, more preferably 5min, and the interval time is preferably 4-6 min, more preferably 5min.
Preferably, when the assembly bacteria degrade the kitchen waste, the treatment temperature of the integrated liquefying device for the kitchen waste is preferably 30-45 ℃, and more preferably 35 ℃.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
The materials used in the examples are as follows:
1. Enrichment medium: 3.0g of beef extract, 10.0g of peptone, 5.0g of NaCl, 1000mL of distilled water and pH 7.2-7.4.
2. Kitchen waste mixed solution: kitchen waste is taken from a dining hall of a college, and after bulk solid waste is removed, the kitchen waste is processed according to the following steps: tap water = 3:5 (mass ratio) mixing the kitchen waste with tap water to obtain the kitchen waste mixed solution.
Example 1
The strain with degradation effects of protein, starch, cellulose, grease and the like is sequenced on a Hiseq 2500 platform. Sequencing to obtain bacterial 16S rRNA gene sequences in each hole, performing Blast comparison at NCBI to obtain a comparison strain of each bacterium, and performing patent program preservation, wherein specific information is as follows.
Wherein the 16S rRNA gene of the bacterium having the number HL-1 has the sequence shown in the sequence 1, and is identified as the Pseudomonas saccharolytica. The bacteria are in a rod shape and are distributed. The colony is white, has smaller shape, round shape, moist, raised surface, transparent, greasy, glossy and neat edge. Bacteria were not capsular, not spore, and gram staining was negative. The registration number of the non-sugar-decomposing pseudomonas (Pseudochrobactrum asaccharolyticum) HL-1 in the common microorganism center of the China Committee for culture Collection of microorganisms is CGMCC No.27442. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Hereinafter referred to as sugar-free pseudo-pale Bai Ganjun HL-1.
Wherein the 16S rRNA gene of the bacterium numbered HL-2 has a sequence as shown in sequence 2 and is identified as Alcaligenes faecalis. Gram-negative bacilli, approximately 0.7-1.0 μm in diameter, and periflagellum movement. Culturing on nitrogen-free plate culture medium with benzoate as carbon source at 30deg.C for 5 days to obtain colony with round shape, smooth surface, raised edge, and 1-1.5mm diameter, and melanin production. Respiratory metabolism, oxidase positivity, and no carbohydrate growth.
Alcaligenes faecalis (ALCALIGENES FAECALIS) HL-2, which has a registration number of CGMCC No.27443 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Hereinafter referred to as Alcaligenes faecalis HL-2.
Wherein the 16S rRNA gene of the bacterium designated HL-3 has a sequence as shown in sequence 3, and is identified as Achromobacter xylosoxidans. Colonies are small, flat, transparent, white, moist, clean-edged, grow faster, gram-negative.
Achromobacter xylosoxidans (Achromobacterxylosoxidans) HL-3 has a registration number of CGMCC No.27444 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Achromobacter xylosoxidans HL-3 are hereinafter abbreviated.
Wherein the 16S rRNA gene of the bacterium having the number HL-4 was identified as Bacillus amyloliquefaciens as shown in the sequence 4. Gram positive bacteria, the colony is pale yellow, has smooth surface, is not moist, has irregular edges and semitransparent, and has the diameter of about 1mm.
Bacillus amyloliquefaciens (Paenibacillus amylolyticus) HL-4 has a registration number of CGMCC No.27445 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Hereinafter, bacillus amyloliquefaciens HL-4 is abbreviated.
Wherein the 16S rRNA gene of the bacterium designated HL-14 has a sequence as shown in SEQ ID No. 5, and is identified as Bacillus cereus. The size is (1 mu m-1.3 mu m) x (3 mu m-5 mu m), spores can be formed, two ends of the thalli are flat, and most of the thalli are arranged in a chain shape; the spores are elliptical and are positioned at one slightly deviated end of the center of the thalli; after the spore formation rate is more than 90% after the spore is cultured for 3d at the temperature of 32 ℃ in nutrient broth, the spore germination can be stimulated by water bath for 5 to 10 minutes at the temperature of 80 to 85 ℃; colony cells grown on glucose watercress contain reddish uncolored pellets, colonies formed on common agar are large, off-white, opaque, rough-surfaced, frosted glass-like or melt-waxy, and the edges are often expanded; the colony on the mannitol vitelline polymyxin agar medium is pink and has white precipitation rings around. The minimum growth temperature is 10-20 ℃, the maximum growth temperature is 35-45 ℃, the bacteria are not propagated below 10 ℃ and above 63 ℃, and the bacteria are easy to die at 65-70 ℃; p H1-2 does not grow, p H-11 can grow, wherein p H4.3.3-9.3 is propagated rapidly; the optimal growth sodium chloride concentration is 1%, and 8% sodium chloride has an inhibition effect on the growth of the plant, and the plant grows well under the condition of no salt.
Bacillus cereus (HL-14) has a registration number of CGMCC No.27446 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Hereinafter, bacillus cereus HL-14 is abbreviated.
Wherein the 16S rRNA gene of the bacterium designated HL-15 has a sequence as shown in sequence 6, and is identified as Sphingobacterium fumigatus. Gram-negative bacteria, colony diameter 2-6mm, yellow, round, middle bulge; round bar shape of the cells, 0.4-0.6 μm wide and 0.8-1.8 μm long, and the cells have no mobility.
Sphingobacterium nicotianae (Sphingobacterium tabacisoli) HL-15, which has a registration number of CGMCC No.27447 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Hereafter abbreviated as Sphingobacterium fumigatus HL-15.
Wherein the 16S rRNA gene of the bacterium designated HL-18 has a sequence as shown in sequence 7, and is identified as Serratia marcescens. The colonies of the strain were essentially convex, opaque in the center, irregular in the edges, and 1 to 2.5mm in size, all producing a red pigment (Panel I-3). Particular colonies (Panel I-1) can be generated on nutrient agar plates in a dendritic state with left-hand rotation; under specific conditions, double-row colonies (plate I-2) can appear which are successively ordered from small to large. The bacterial cells are gram-negative short bacillus, the size is (1-1.3) mu m x (0.7-1.0) mu m, and the bacterial cells are periphytic flagellum, dynamic, non-capsular and non-spore.
Serratia marcescens (SERRATIA MARCESCENS) HL-18 has a registration number of CGMCC No.27448 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Serratia marcescens HL-18 is hereinafter abbreviated.
Wherein the 16S rRNA gene of the bacterium numbered HL-20 was identified as stenotrophomonas maltophilia as shown in SEQ ID NO 8. Strong ammonia smell exists on the blood plate, and the blood plate is beta-hemolyzed; on nutrient agar, there is grey yellow pigment or no pigment, the colony is needle tip, the diameter is 0.5-1 mm, and the center is raised.
Stenotrophomonas maltophilia (Stenotrophomonas maltophilia) HL-20 has a registration number of CGMCC No.27449 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Hereinafter referred to as stenotrophomonas maltophilia HL-20.
Wherein the 16S rRNA gene of the bacterium designated HL-25 has the sequence shown in SEQ ID No. 9, and is identified as Sphingobacterium humicola. The colony is circular, milky yellow, opaque, smooth and moist in surface, slightly convex in center, regular in edge and free of halo.
Sphingobacterium humus (Sphingobacterium humi) HL-25 with a registration number of CGMCC No.27450 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. Hereinafter, sphingobacterium humus HL-25.
Wherein the 16S rRNA gene of the bacterium designated HL-26 has a sequence as shown in SEQ ID No. 10, and is identified as Sphingobacterium aquaticum. The cells are in a short rod shape, the width is 0.2-0.7 mu m, and the length is 1.0-2.5 mu m; gram-negative bacteria, do not move and do not produce spores.
Sphingobacterium hydrobromide (Sphingobacterium mizutaii) HL-26, which has a registration number of CGMCC No.27451 in the China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No. 1 and No. 3 in the Korean area of Beijing city. The Sphingobacterium aquaticum HL-26 is hereinafter abbreviated.
Wherein the 16S rRNA gene of the bacterium designated HL-27 has a sequence as shown in sequence 11, and is identified as Bacillus subtilis. The single cell is 0.7-0.8X2-3 microns and the coloring is uniform. Without capsule, the flagellum can be grown and can move. Gram positive bacteria can form endogenous stress-resistant spores, the spores are 0.6-0.9X1.0-1.5 microns, ellipse is columnar, and the spores are positioned in the center or slightly deviated from the bacteria, so that the bacteria do not expand after the spores are formed. The growth and propagation speed is high, the colony surface is rough and opaque, the pollution is white or yellowish, and when the colony grows in a liquid culture medium, the colony surface often forms wrinkling, so the colony is an aerobic bacterium.
Bacillus subtilis (Bacillus subtilis) HL-27 with registration number of CGMCC No.27452 in China general microbiological culture Collection center. The strain is preserved in China general microbiological culture collection center (CGMCC) for 5 months and 23 days in 2023, and the preservation address is North Star Xiu No.1 and No. 3 in the Korean area of Beijing city. Hereinafter, bacillus subtilis HL-27 is abbreviated.
Example 2
The 11 strains in example 1 were respectively expanded to a liquid medium, and the strains were inoculated into a protein, starch, cellulose and lipid screening medium according to an initial biomass of 0.05g/L, the cellulose, protein, starch and lipid contents of 0 and 6d were respectively measured, and the degradation rates of the different strains on the above substances were calculated, and the results are shown in FIG. 1.
After screening out strains capable of efficiently degrading various substances, designing orthogonal experiments aiming at degradation of four substances to obtain optimal composite bacterial systems with different degradation functions, and mixing the composite bacterial systems to obtain final composite bacteria, wherein the final composite bacteria comprise 11 strains in example 1 with equal biomass ratio. The composite bacteria were added to the four kinds of screening media, and after 6d of cultivation, the degradation rates of the four kinds of substances were measured, and the degradation effect of the composite bacteria was evaluated, and the results are shown in FIG. 2. In FIG. 1, strains corresponding to different numbers on the abscissa are shown in Table 1.
TABLE 1 strain numbering and name
As can be seen from the above results, the maximum degradation rates of a single strain on protein, cellulose, grease and starch were 35.59%, 45.50%, 94.34%, 99.71%, and the corresponding strains were SERRATIA MARCESCENS (Serratia marcescens), sphingobacterium mizutaii (Sphingobacterium hydrobromide), paenibacillus amylolyticus (Bacillus amyloliquefaciens), paenibacillus amylolyticus (Bacillus amyloliquefaciens), respectively. As shown in FIG. 2, the degradation rates of the complex bacteria obtained by the orthogonal experiment on protein, starch, grease and cellulose are 24.53%, 96.51%, 63.72% and 34.14%, respectively.
Example 2
Mixing 30g of kitchen waste with 50g of tap water, and adding into a 100mL conical flask; adding the composite bacteria into the kitchen waste mixed solution according to the biomass of 0.22g/L (the initial biomass of each bacteria is 0.02 g/L); placing the conical flask in a constant temperature water bath oscillator at 30 ℃ with the rotating speed of 130rpm/min; sampling at the 0 th day and the 6 th day, measuring the contents of protein, starch, cellulose and grease in the kitchen waste mixed solution, and measuring the weight and the volume of the kitchen waste.
In the embodiment 2, the compound bacteria can effectively remove 85.21%, 59.00%, 94.34% and 27.11% of cellulose, grease, starch and protein in the kitchen waste mixed solution. The composite bacteria can also liquefy part of kitchen waste, effectively reduce the volume and the weight of the kitchen waste, and the weight reduction rate before and after liquefaction are 73.00% and 75.59% respectively. The results are shown in FIG. 3.
Example 3
The invention aims to solve the problem of degradation and reduction of kitchen waste, and provides a pilot scale process and a pilot scale method for degrading the kitchen waste by utilizing composite bacteria. Composite bacteria are added into kitchen waste biochemical treatment aerobic fermentation equipment of staff restaurants of a certain municipal company in Beijing, so that remarkable degradation and reduction of the kitchen waste are successfully realized. By adjusting the operation parameters of the equipment, the effect of the compound bacteria is exerted to the maximum extent. Within 7 days, the treatment equipment treats 509.20 kg of kitchen waste, and the waste decrement rate reaches 99.78% after degradation of the composite bacteria. The calculation formula of the kitchen waste decrement rate is as follows: kitchen waste reduction rate=1- (1.1/509.20) =99.78% (note: parameter 1.1 is mass of undegraded cellulose in the waste treatment equipment after 7 days).
The composite bacteria of the invention are applied to a pilot-scale process for one week, and the specific operation steps are as follows:
(1) Performing expansion culture of the composite bacterial liquid in a laboratory;
(2) Adding the composite bacterial liquid into garbage treatment equipment, and simultaneously adding bacterial attachment materials;
(3) Measuring the feeding amount of garbage each time, observing the height change condition of kitchen garbage in the equipment, collecting effluent before and after garbage feeding, and detecting the effluent quality.
The composite bacteria disclosed by the invention are applied to a pilot-scale process, and the result shows that after one-night degradation (14 h), the height of the garbage is obviously reduced, the maximum reduction rate of the height of the garbage can reach 14.66%, the height of the garbage shows a stable fluctuation trend in the same day, and the effective reduction of kitchen garbage is realized. Meanwhile, after the composite bacteria degrade kitchen waste, the COD removal rate of the obtained effluent liquefied liquid can reach 98.19% at the highest. The parameters of the garbage treatment equipment are optimized, the performance of the composite bacterial liquid is obviously improved, the viscosity change of the garbage liquefied liquid is reflected, and the degradation performance of garbage is also improved.
The results are shown in FIGS. 4-6, and FIG. 4 shows the garbage height before and after garbage feeding in 1 day in the pilot plant of example 2 (the abscissa 1-8 in the figure shows different sampling points for collecting water. 1 before breakfast garbage feeding, 2 after breakfast garbage feeding, 3, morning, 4 before lunch garbage feeding, 5 after lunch garbage feeding, 6 afternoon, 7 before supper garbage feeding, 8 after supper garbage feeding); FIG. 5 shows the reduction rate of the height of kitchen waste after being degraded by the composite bacteria for one night (14 h) (the abscissa of the figure is 1-6 respectively showing the height of the waste versus the time point); FIG. 6 shows the COD removal rate of the kitchen waste liquefaction liquid effluent after being degraded by the composite bacteria for one night (14 h) (the abscissa of the graph 1-6 is the COD removal rate versus time respectively).
In conclusion, the composite bacteria can effectively degrade and liquefy the kitchen waste.
According to the embodiment, the invention provides the composite bacteria for degrading the liquefied kitchen waste and the application thereof, the composite bacteria comprise 11 bacteria, namely, pseudomonas solanacearum, alcaligenes faecalis, paenibacillus amyloliquefaciens, serratia marcescens, sphingosine-helper, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingosine-fumigatus, stenotrophomonas maltophilia and sphingosine-humate, the composite bacteria with the seed retention numbers of Pseudochrobactrum asaccharolyticum、Alcaligenesfaecalis、Paenibacillus amylolyticus、Serratia marcescens、Sphingobacterium mizutaii、Bacillus subtilis、Achromobacter xylosoxidans、Bacillus cereus、Sphingobacterium tabacisoli、Stenotrophomonas maltophilia、Sphingobacterium humi. are CGMCCNO.27442、CGMCCNO.27443、CGMCCNO.27445、CGMCCNO.27448、CGMCCNO.27451、CGMCCNO.27452、CGMCCNO.27444、CGMCCNO.27446、CGMCCNO.27447、CGMCCNO.27449、CGMCCNO.27450., the composite bacteria can still realize good continuous degradation effect on the kitchen waste in practical pilot-plant test application, and the waste reduction rate in one week can reach 99.78%. The culture condition of the composite bacteria is easy to realize and control, can effectively degrade and liquefy the kitchen waste, and has great significance for the efficient treatment of the kitchen waste.
The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
Claims (10)
1. An application of a strain combination in liquefying and degrading kitchen waste, wherein the strain combination comprises Sphingobacterium fumigatus (Sphingobacterium tabacisoli).
2. The use according to claim 1, wherein the combination of species comprises Achromobacter xylosoxidans (Achromobacterxylosoxidans), serratia marcescens (SERRATIA MARCESCENS), sphingobacterium aquaticum (Sphingobacterium mizutaii) and Sphingobacterium humicola (Sphingobacterium humi), and Sphingobacterium fumigatus (Sphingobacterium tabacisoli).
3. The use according to claim 2, characterized in that the combination of strains consists of pseudomonas saccharolytica (Pseudochrobactrum asaccharolyticum), alcaligenes faecalis (ALCALIGENESFAECALIS), paenibacillus amyloliquefaciens (Paenibacillus amylolyticus), serratia marcescens (SERRATIA MARCESCENS), sphingobacterium aquaticum (Sphingobacterium mizutaii), bacillus subtilis (Bacillus subtilis), achromobacter xylosoxidans (Achromobacterxylosoxidans), bacillus cereus (Bacillus cereus), sphingobacterium fumigatus (Sphingobacterium tabacisoli), stenotrophomonas maltophilia (Stenotrophomonas maltophilia) and sphingobacterium humicola (Sphingobacterium humi).
4. A microbial inoculum for liquefying and degrading kitchen waste, which is characterized in that the active ingredients of the microbial inoculum are the strain combinations in any one of claims 1-3.
5. The microbial inoculum according to claim 4, wherein the microbial inoculum is formed by mixing a single-strain enriched solution of pseudomonas saccharolytica, alcaligenes faecalis, paenibacillus amyloliquefaciens, serratia marcescens, sphingobacterium aquaticum, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingobacterium fumigatus, stenotrophomonas maltophilia and sphingobacterium humicola.
6. The microbial inoculum of claim 5, wherein the single-fungus enrichment medium is a microbial inoculum obtained by inoculating a strain into an enrichment culture medium, each liter of the enrichment culture medium comprises 3.0g of beef extract, 10.0g of peptone and 5.0g of NaCl, and the pH of the enrichment culture medium is 7.2-7.4.
7. The microbial inoculum according to claim 5, wherein the biomass ratio of pseudomonas saccharolytica, alcaligenes faecalis, paenibacillus amyloliquefaciens, serratia marcescens, sphingobacterium aquaticum, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingobacterium fumigatus, stenotrophomonas maltophilia and sphingobacterium humicola is 1;1:1:1:1:1;1:1:1:1:1.
8. A combination of species as claimed in any one of claims 1 to 3.
9. The kitchen waste liquefying and degrading method is characterized in that the compound bacteria in any one of claims 1-3 or the microbial inoculum in any one of claims 4-6 is used for inoculating into the kitchen waste mixed liquid.
10. The method according to claim 9, wherein after inoculation into the mixed solution of kitchen waste, the initial biomass of the pseudomonas saccharolytica, alcaligenes faecalis, paenibacillus amyloliquefaciens, serratia marcescens, sphingobacterium aquaticum, bacillus subtilis, achromobacter xylosoxidans, bacillus cereus, sphingobacterium fumigatus, stenotrophomonas maltophilia and sphingobacterium humicola is 0.01-0.03 g/L, and the total biomass in the mixed solution of kitchen waste is 0.11-0.33 g/L.
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