A combination of superior bacteria for liquefying and degrading kitchen waste and its application

Abstract

The present invention provides a combination of superior bacteria for liquefying and degrading food waste and its application. The combination includes Pseudomonas aglycones, Bacillus alcaligenes faecalis, Bacillus amyloliquefaciens, Serratia marcescens, Sphingobacillus suis, Bacillus subtilis, Achromobacter xylosoxidans, Bacillus cereus, Sphingobacillus fusoni, Oligotrophomonas maltophilia, and Sphingobacillus humus. The combination of bacteria can effectively reduce the amount of food waste, and the maximum reduction can be 14.66%, and the COD removal rate can reach 98.19%. The combination of bacteria of the present invention can efficiently degrade and liquefy food waste, and can achieve a high reduction rate and liquefaction effect, which is conducive to the absorption and utilization of nutrients in the subsequent food waste biological resource conversion process.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a combination of superior bacteria for liquefying and degrading kitchen waste and applications thereof. Background Art

[0002] With the rapid development of the catering industry, the amount of food waste generated has increased rapidly, with the characteristics of large output and high concentration of pollutants. If not properly handled, it will cause serious pollution to the environment and affect the normal life of some organisms. Traditional food waste treatment methods have defects. Biological treatment methods are low-cost and high-efficiency, but microorganisms often cannot directly utilize macromolecules in food waste. Therefore, liquefying food waste can significantly improve the utilization rate of nutrients in sewage by microorganisms, which is conducive to subsequent purification and resource utilization. Developing a method that can quickly degrade liquefied food waste is the key to efficient treatment of food waste. Summary of the invention

[0003] The purpose of the present invention is to provide a combination of superior bacteria for liquefying and degrading kitchen waste and its application.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0005] The invention provides a dominant bacteria combination, wherein the combined bacteria include Pseudochrobactrum asaccharolyticum, Alcaligenes faecalis, Bacillus amylolyticus, Serratia marcescens, Sphingobacterium suis, Bacillus subtilis, Achromobacter xylosoxidans, Bacillus cereus, Sphingobacterium spp. in tobacco fields, Stenotrophomonas maltophilia, and Sphingobacterium humi in Latin. The combined bacteria include Pseudochrobactrum asaccharolyticum, Alcaligenes faecalis, Paenibacillus amylolyticus, Serratia marcescens, Sphingobacterium mizutaii, Bacillus subtilis, Achromobacter xylosoxidans, Bacillus cereus, Sphingobacterium tabacisoli, Stenotrophomonas maltophilia, and Sphingobacterium humi.

[0006] In the present invention, various strains in the combined bacteria can be obtained by conventional separation and screening means.

[0007] The present invention also provides application of the combined bacteria in degrading liquefied food waste.

[0008] Preferably, the bacterial combination is formed by mixing single-bacteria enrichment solutions of 11 bacteria.

[0009] The single bacteria enrichment culture medium comprises the following components: 3.0 g / L beef extract, 10.0 g / L peptone, 5.0 g / L NaCl, and pH 7.2-7.4.

[0010] Preferably, the initial biomass of the 11 bacteria in the combined strains is 0.01-0.03 g / L, and the total initial biomass after mixing is 0.11-0.33 g / L.

[0011] As a preference, when the combined bacteria degrades the kitchen waste, the tap water spraying time of the kitchen waste integrated liquefaction device is 120 seconds, and the interval time is 10 minutes.

[0012] As a preference, when the combined bacteria degrades food waste, the dosing time of the bacterial solution in the food waste integrated liquefaction device is 20 seconds and the interval time is 3 hours.

[0013] As a preference, when the combined bacteria degrades the food waste, the garbage stirring time of the integrated food waste liquefaction device is 5 minutes, and the interval time is 5 minutes.

[0014] Preferably, when the combined bacteria degrades the food waste, the processing temperature of the integrated food waste liquefaction device is 30-45°C.

[0015] Collection Instructions

[0016] Biomaterials 1

[0017] Classification and nomenclature of biological materials: Pseudochrobactrum asaccharolyticum .

[0018] The strain number of biological material: HL-1.

[0019] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0020] The abbreviation of the depository of biological materials is: CGMCC.

[0021] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0022] Deposit date of biological material: May 23, 2023.

[0023] The registration number of the Collection Center for Biological Materials is: CGMCC No.27442.

[0024] Biomaterials 2

[0025] Classification and nomenclature of biological materials: Alcaligenes faecalis faecalis).

[0026] The strain number of biological material: HL-2.

[0027] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0028] The abbreviation of the depository of biological materials is: CGMCC.

[0029] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0030] Deposit date of biological material: May 23, 2023.

[0031] The registration number of the Collection Center for Biological Materials is: CGMCC No.27443.

[0032] Biomaterials 3

[0033] Classification and nomenclature of biological materials: Achromobacter xylosoxidans xylosoxidans).

[0034] The strain number of biological material: HL-3.

[0035] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0036] The abbreviation of the depository of biological materials is: CGMCC.

[0037] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0038] Deposit date of biological material: May 23, 2023.

[0039] The registration number of the Collection Center for Biological Materials is: CGMCC No.27444.

[0040] Biomaterials 4

[0041] Classification and nomenclature of biological materials: Paenibacillus amyloliquefaciens amylolyticus).

[0042] The strain number of biological material: HL-4.

[0043] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0044] The abbreviation of the depository of biological materials is: CGMCC.

[0045] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0046] Deposit date of biological material: May 23, 2023.

[0047] The registration number of the Collection Center for Biological Materials is: CGMCC No.27445.

[0048] Biomaterials 5

[0049] Classification and nomenclature of biological materials: Bacillus cereus.

[0050] The strain number of biological material: HL-14.

[0051] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0052] The abbreviation of the depository of biological materials is: CGMCC.

[0053] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0054] Deposit date of biological material: May 23, 2023.

[0055] The registration number of the Collection Center for Biological Materials is: CGMCC No.27446.

[0056] Biomaterials 6

[0057] Classification and naming of biological materials: Sphingobacterium tabacisoli).

[0058] The strain number of biological material: HL-15.

[0059] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0060] The abbreviation of the depository of biological materials is: CGMCC.

[0061] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0062] Deposit date of biological material: May 23, 2023.

[0063] The registration number of the Collection Center for Biological Materials is: CGMCC No.27447.

[0064] Biomaterials 7

[0065] Classification and nomenclature of biological materials: Serratia marcescens.

[0066] The strain number of biological material: HL-18.

[0067] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0068] The abbreviation of the depository of biological materials is: CGMCC.

[0069] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0070] Deposit date of biological material: May 23, 2023.

[0071] The registration number of the Collection Center for Biological Materials is: CGMCC No.27448.

[0072] Biomaterials 8

[0073] Classification and nomenclature of biological materials: Stenotrophomonas maltophilia maltophilia).

[0074] The strain number of biological material: HL-20.

[0075] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0076] The abbreviation of the depository of biological materials is: CGMCC.

[0077] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0078] Deposit date of biological material: May 23, 2023.

[0079] The registration number of the Collection Center for Biological Materials is: CGMCC No.27449.

[0080] Biomaterials 9

[0081] Classification and nomenclature of biological materials: Sphingobacterium humi.

[0082] The strain number of biological material: HL-25.

[0083] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0084] The abbreviation of the depository of biological materials is: CGMCC.

[0085] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0086] Deposit date of biological material: May 23, 2023.

[0087] The registration number of the Collection Center for Biological Materials is: CGMCC No.27450.

[0088] Biomaterials 10

[0089] Classification and naming of biological materials: Sphingobacterium mizutaii).

[0090] The strain number of biological material: HL-26.

[0091] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0092] The abbreviation of the depository of biological materials is: CGMCC.

[0093] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0094] Deposit date of biological material: May 23, 2023.

[0095] The registration number of the Collection Center for Biological Materials is: CGMCC No.27451.

[0096] Biomaterials11

[0097] Classification and nomenclature of biological materials: Bacillus subtilis.

[0098] The strain number of biological material: HL-27.

[0099] Name of the depository unit of biological materials: General Microbiology Center of China Microbiological Culture Collection Administration.

[0100] The abbreviation of the depository of biological materials is: CGMCC.

[0101] Address of the depository of biological materials: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China Institute, Postal Code: 100101.

[0102] Deposit date of biological material: May 23, 2023.

[0103] The registration number of the Collection Center for Biological Materials is: CGMCC No.27452.

[0104] The invention provides a composite bacteria for degrading liquefied kitchen waste and application thereof. The composite bacteria comprises 11 kinds of bacteria, namely, Pseudochrobactrum asaccharolyticum, Alcaligenes faecalis, Paenibacillus amylolyticus, Serratia marcescens, Sphingobacterium suis, Bacillus subtilis, Achromobacter xylosoxidans, Bacillus cereus, Sphingobacterium spp. in tobacco fields, Stenotrophomonas maltophilia and Sphingobacterium humi in Latin. The 11 kinds of bacteria are Pseudochrobactrum asaccharolyticum, Alcaligenes faecalis, Paenibacillus amylolyticus, Serratia marcescens, Sphingobacterium mizutaii, Bacillus subtilis, Achromobacterxylosoxidans, Bacillus cereus, Sphingobacterium tabacisoli, Stenotrophomonas maltophilia and Sphingobacterium humi. The seed numbers are CGMCCNO.27442, CGMCCNO.27443, CGMCCNO.27445, CGMCCNO.27448, CGMCCNO.27451, CGMCCNO.27452, CGMCCNO.27444, CGMCCNO.27446, CGMCCNO.27447, CGMCCNO.27449 and CGMCCNO.27450 respectively. The composite bacteria of the present invention are used to treat kitchen waste. In actual pilot applications, the composite bacteria can still achieve a good and continuous degradation effect on kitchen waste, and the waste reduction rate can reach 99.78% within one week. The culture conditions of the composite bacteria are easy to realize and control, and the composite bacteria can effectively degrade and liquefy kitchen waste, which is of great significance for the efficient treatment of kitchen waste. BRIEF DESCRIPTION OF THE DRAWINGS

[0105] Figure 1The maximum degradation rate of protein (a), cellulose (b), oil (c) and starch (d) by a single bacterial species in Example 2.

[0106] Figure 2 The degradation rates of protein, starch, oil and cellulose by the composite bacteria (11 species) in Example 2 are shown in FIG.

[0107] Figure 3 The changes in various components after the mixed liquid of kitchen waste was treated with composite bacteria for 6 days in Example 3.

[0108] Figure 4 The height change of garbage before and after garbage feeding in the pilot process equipment of Example 4 within one day (the horizontal axes 1-6 in the figure are different sampling points for collecting effluent. 1: before breakfast garbage feeding; 2: after breakfast garbage feeding; 3: before lunch garbage feeding; 4: after lunch garbage feeding; 5: before dinner garbage feeding; 6: after dinner garbage feeding)

[0109] Figure 5 It is the reduction rate of garbage height after 4 examples of Chinese food waste were degraded by the composite bacteria of the present invention for one night (14h) (the horizontal axes 1-6 in the figure are the garbage height comparison time points respectively).

[0110] Figure 6 The COD removal rate of the effluent from the liquefied food waste after one night (14 hours) of composite bacteria degradation in 4 cases (the horizontal axes 1-6 in the figure are COD removal rate versus time, respectively). DETAILED DESCRIPTION

[0111] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0112] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0113] The invention provides a combination of superior bacteria for liquefying and degrading kitchen waste and application thereof. The combined bacteria include Pseudochrobactrum asaccharolyticum, Alcaligenes faecalis, Bacillus amylolyticus, Serratia marcescens, Sphingobacterium suis, Bacillus subtilis, Achromobacter xylosoxidans, Bacillus cereus, Sphingobacterium spp. in tobacco fields, Stenotrophomonas maltophilia and Sphingobacterium humi in Latin. The bacteria are Pseudochrobactrum asaccharolyticum, Alcaligenes faecalis, Paenibacillus amylolyticus, Serratia marcescens, Sphingobacterium mizutaii, Bacillus subtilis, Achromobacter xylosoxidans, Bacillus cereus, Sphingobacterium tabacisoli, Stenotrophomonas maltophilia and Sphingobacterium humi.

[0114] The present invention also provides application of the combined bacteria in degrading liquefied food waste.

[0115] In the present invention, the assembled bacteria are inoculated into an integrated food waste liquefaction treatment device.

[0116] In the present invention, the initial biomass of the 11 bacteria in the combined bacteria is the same, preferably 0.01-0.03 g / L, more preferably 0.02 g / L, and the total initial biomass is 0.11-0.33 g / L, more preferably 0.22 g / L.

[0117] In the present invention, when the combined bacteria degrades food waste, the tap water spraying time of the food waste integrated liquefaction device is preferably 110 to 130 seconds, more preferably 120 seconds, and the interval time is preferably 5 to 15 minutes, more preferably 10 minutes.

[0118] Preferably, when the combined bacteria degrades food waste, the dosing time of the bacteria solution of the integrated food waste liquefaction device is preferably 10 to 30 seconds, more preferably 20 seconds, and the interval time is preferably 2 to 3 hours, more preferably 3 hours.

[0119] Preferably, when the combined bacteria degrades food waste, the garbage stirring time of the integrated food waste liquefaction device is preferably 4 to 6 minutes, more preferably 5 minutes, and the interval time is preferably 4 to 6 minutes, more preferably 5 minutes.

[0120] Preferably, when the combined bacteria degrades food waste, the processing temperature of the integrated food waste liquefaction device is preferably 30-45°C, and more preferably 35°C.

[0121] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0122] The materials used in the examples are as follows:

[0123] 1. Enrichment medium: 3.0 g beef extract, 10.0 g peptone, 5.0 g NaCl, 1000 mL distilled water, pH 7.2-7.4.

[0124] 2. Kitchen waste mixture: The kitchen waste was taken from a university cafeteria. After removing large pieces of solid waste, the kitchen waste and tap water were mixed in a ratio of kitchen waste: tap water = 3:5 (mass ratio) to obtain a kitchen waste mixture.

[0125] Example 1

[0126] The strains screened out with protein, starch, cellulose and oil degradation effects were sequenced on the HiSeq 2500 platform. The 16S rRNA gene sequence of the bacteria in each well was sequenced and compared by Blast in NCBI to obtain the comparison strains of each bacterium and deposited in the patent procedure. The specific information is as follows.

[0127] The sequence of the 16S rRNA gene of the bacterium numbered HL-1 is shown in Sequence 1, and it is identified as Pseudochrobactrum asaccharolyticum. The bacteria are rod-shaped and dispersed. The colonies are white, small in shape, round, moist, with raised surfaces, transparent, oily, shiny, and neatly marginated. The bacteria have no capsule, no spores, and Gram staining is negative. Pseudochrobactrum asaccharolyticum HL-1, its registration number in the General Microbiological Center of the China Microbiological Culture Collection Administration is CGMCC No. 27442. The strain was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration (referred to as CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. hereinafter referred to as Pseudochrobactrum asaccharolyticum HL-1.

[0128] The sequence of the 16S rRNA gene of the bacterium numbered HL-2 is shown in Sequence 2, and it is identified as Alcaligenes faecalis. It is a Gram-negative bacillus with a diameter of about 0.7-1.0 μm and a peritrichous flagella. On a nitrogen-free plate medium with benzoate as a carbon source, cultured at 30°C for 5 days, the colonies are round, with a smooth surface, protrusions, neat edges, and a diameter of 1-1.5 mm, and produce melanin. It has respiratory metabolism, is oxidase-positive, and does not use sugars for growth.

[0129] Alcaligenes faecalis HL-2, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27443. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Alcaligenes faecalis HL-2.

[0130] The sequence of the 16S rRNA gene of the bacterium numbered HL-3 is shown in Sequence 3, and it is identified as Achromobacter xylosoxidans. The colony is small, flat, transparent, white, with a moist surface, neat edges, fast growth, and Gram-negative.

[0131] Achromobacter xylosoxidans HL-3, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27444. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Achromobacter xylosoxidans HL-3.

[0132] The sequence of the 16S rRNA gene of the bacterium numbered HL-4 is shown in Sequence 4, and it is identified as Bacillus amyloliquefaciens. It is a Gram-positive bacterium, with a light yellow colony, a smooth surface, no moisture, irregular edges, and a translucent colony size of about 1 mm.

[0133] Paenibacillus amylolyticus HL-4, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27445. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Paenibacillus amylolyticus HL-4.

[0134] The sequence of the 16S rRNA gene of the bacterium numbered HL-14 is shown in Sequence 5, and it is identified as Bacillus cereus. The size is (1μm~1.3μm)×(3μm~5μm), and it can form spores. The two ends of the bacteria are relatively flat, and most of them are arranged in chains; its spores are oval and located slightly to one end of the center of the bacteria; after culturing in nutrient broth at 32℃ for 3 days, the spore formation rate is above 90%, and 80℃~85℃ water bath for 5min~10min can stimulate spore germination; the colony cells grown on glucose agar contain small balls that are not colored by fuchsin, and the colonies formed on ordinary agar are larger, grayish white, opaque, with a rough surface like frosted glass or melted wax, and the edges are often expanded; the colonies on mannitol egg yolk polymyxin agar medium are pink with a white precipitation ring around them. The lowest growth temperature is 10℃~20℃, the highest growth temperature is 35℃~45℃, it will not reproduce below 10℃ and above 63℃, and the bacteria are easy to die at 65℃~70℃; it will not grow at pH 1~2, and it can grow at pH 2~11, among which, it reproduces rapidly at pH 4.3~9.3; the most suitable growth sodium chloride concentration is 1%, 8% sodium chloride has an inhibitory effect on its growth, and it grows well in the absence of salt.

[0135] Bacillus cereus HL-14, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27446. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Bacillus cereus HL-14.

[0136] The sequence of the 16S rRNA gene of the bacterium numbered HL-15 is shown in Sequence 6, and it is identified as Sphingobacterium nicotianae. It is a Gram-negative bacterium, with a colony diameter of 2-6 mm, yellow, round, and convex in the middle; the cells are rod-shaped, 0.4-0.6 μm wide and 0.8-1.8 μm long, and the cells are non-motile.

[0137] Sphingobacterium tabacisoli HL-15, whose registration number in the General Microbiological Center of China Microorganism Culture Collection Administration Committee is CGMCC No.27447. The strain was deposited in the General Microbiological Center of China Microorganism Culture Collection Administration Committee (referred to as CGMCC) on May 23, 2023, and the deposit address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Sphingobacterium tabacisoli HL-15.

[0138] The sequence of the 16S rRNA gene of the bacterium numbered HL-18 is shown in Sequence 7, and it is identified as Serratia marcescens. The colonies of the strain are basically convex, opaque in the center, irregular at the edges, 1 to 2.5 mm in size, and all produce red pigment (Plate Ⅰ-3). . On the nutrient agar plate, it can produce special colonies that are dendritic and left-handed (Plate Ⅰ-1); when cultured under special conditions, double-row colonies that gradually increase in size and are arranged in an orderly manner can appear (Plate Ⅰ-2). The bacteria are Gram-negative short rods, with a size of (1 to 1.3) μm × (0.7 to 1.0) μm, flagella around the peritrichous, motile, without capsules, and without spores.

[0139] Serratia marcescens HL-18, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27448. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Serratia marcescens HL-18.

[0140] The sequence of the 16S rRNA gene of the bacterium numbered HL-20 is shown in Sequence 8, and it is identified as Stenotrophomonas maltophilia. It has a strong ammonia smell on the blood agar plate and is β-hemolytic; it shows gray-yellow pigment or no pigment on nutrient agar, and the colony is needle-shaped, with a diameter of 0.5mm to 1mm and a central protrusion.

[0141] Stenotrophomonas maltophilia HL-20, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27449. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Stenotrophomonas maltophilia HL-20.

[0142] The sequence of the 16S rRNA gene of the bacterium numbered HL-25 is shown in Sequence 9, and it is identified as Sphingobacterium humum. The colony is round, milky yellow and opaque, with a smooth and moist surface, a slightly convex center, regular edges, and no halo.

[0143] Sphingobacterium humi HL-25, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27450. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Sphingobacterium humi HL-25.

[0144] The sequence of the 16S rRNA gene of the bacterium numbered HL-26 is shown in sequence 10, and it is identified as Sphingobacterium waterbacillus. It is light yellow, smooth, convex, and has a single colony morphology with neat edges. The cells are short rod-shaped, 0.2-0.7 μm wide and 1.0-2.5 μm long. It is a Gram-negative bacterium that is non-motile and does not produce spores.

[0145] Sphingobacterium mizutaii HL-26, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27451. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Sphingobacterium mizutaii HL-26.

[0146] The sequence of the 16S rRNA gene of the bacterium numbered HL-27 is shown in sequence 11, and it is identified as Bacillus subtilis. A single cell is 0.7-0.8×2-3 microns and is evenly colored. It has no capsule, flagella around it, and can move. It is a Gram-positive bacterium that can form endogenous stress-resistant spores, which are 0.6-0.9×1.0-1.5 microns, elliptical to columnar, located in the center of the bacterium or slightly off it. The bacterium does not swell after spore formation. It grows and reproduces quickly, the surface of the colony is rough and opaque, dirty white or slightly yellow, and often forms wrinkles when grown in liquid culture medium. It is an aerobic bacterium.

[0147] Bacillus subtilis HL-27, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection Administration is CGMCC No.27452. The strain was deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration (CGMCC) on May 23, 2023, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Hereinafter referred to as Bacillus subtilis HL-27.

[0148] Example 2

[0149] The 11 strains in Example 1 were cultured in liquid culture medium, and inoculated into protein, starch, cellulose and oil screening medium at an initial biomass of 0.05 g / L. The cellulose, protein, starch and oil contents on the 0th and 6th day were measured, respectively, and the degradation rates of the above substances by different strains were calculated. The results are shown in FIG. Figure 1 shown.

[0150] After screening out strains that can efficiently degrade various substances, an orthogonal experiment was designed for the degradation of the four substances to obtain the best composite bacterial system with different degradation functions, and then the composite bacterial systems were mixed to obtain the final composite bacteria, which contained the 11 strains in Example 1 with equal biomass ratios. The composite bacteria were added to the four screening culture media, and the degradation rates of the four substances were measured after 6 days of culture to evaluate the degradation effect of the composite bacteria. The results are as follows: Figure 2 shown. Figure 1 In the figure, the bacterial species corresponding to different numbers on the horizontal axis are shown in Table 1.

[0151] Table 1 Bacterial strain number and name

[0152]

[0153] From the above results, it can be seen that the maximum degradation rates of protein, cellulose, oil and starch by a single strain are 35.59%, 45.50%, 94.34% and 99.71%, respectively, and the corresponding strains are Serratia marcescens, Sphingobacterium mizutaii, Paenibacillus amylolyticus and Paenibacillus amylolyticus. Figure 2 As shown, the degradation rates of protein, starch, oil and cellulose by the composite bacteria obtained through orthogonal experiments were 24.53%, 96.51%, 63.72% and 34.14%, respectively.

[0154] Example 2

[0155] Mix 30g of food waste and 50g of tap water and add them to a 100mL conical flask; add the composite bacteria to the food waste mixture at a biomass of 0.22g / L (the initial biomass of each bacteria is 0.02g / L); place the conical flask in a constant temperature water bath oscillator at 30°C and a rotation speed of 130rpm / min; take samples on the 0th and 6th days to determine the contents of protein, starch, cellulose and oil in the food waste mixture, and determine the weight and volume of the food waste.

[0156] In Example 2, the composite bacteria can effectively remove 85.21%, 59.00%, 94.34%, and 27.11% of cellulose, oil, starch, and protein in the mixed liquid of food waste. The composite bacteria can also liquefy part of the food waste, effectively reducing its volume and weight. The weight loss rate and reduction rate before and after liquefaction are 73.00% and 75.59%, respectively. The results are shown in Figure 3 .

[0157] Example 3

[0158] The present invention aims to solve the problem of reducing the amount of food waste by degradation, and provides a pilot process and method for degrading food waste by using composite bacteria. Composite bacteria were added to the aerobic fermentation equipment for biochemical treatment of food waste in the employee restaurant of a municipal company in Beijing, and the significant degradation and reduction of food waste were successfully achieved. By adjusting the operating parameters of the equipment, the effect of the composite bacteria was maximized. Within 7 days, the treatment equipment treated 509.20 kilograms of food waste, and the waste reduction rate reached 99.78% after degradation by the composite bacteria. The calculation formula of the food waste reduction rate is as follows: food waste reduction rate = 1-(1.1 / 509.20) = 99.78% (Note: parameter 1.1 is the mass of undegraded cellulose in the waste treatment equipment after 7 days).

[0159] The composite bacteria of the present invention was subjected to a one-week pilot process application, and the specific operation steps were as follows:

[0160] (1) Expanding the culture of the composite bacterial solution in the laboratory;

[0161] (2) adding the composite bacterial solution into the garbage treatment equipment and adding the bacterial attachment material;

[0162] (3) Measure the amount of garbage fed each time, observe the height change of the food waste in the equipment, collect the effluent before and after the garbage feeding, and test the effluent water quality.

[0163] The composite bacteria of the present invention are applied to the pilot process, and the results show that after one night of degradation (14h), the height of the garbage is significantly reduced, and the maximum reduction rate of the garbage height can reach 14.66%. The height of the garbage shows a stable fluctuation trend within the same day, and the effective reduction of the amount of food waste is achieved. At the same time, after the composite bacteria of the present invention degrade the food waste, the COD removal rate of the effluent liquefied liquid can reach up to 98.19%. The parameters of the garbage treatment equipment are optimized, and the performance of the composite bacteria liquid is significantly improved, which is reflected in the viscosity change of the garbage liquefied liquid, and the degradation performance of the garbage is also improved accordingly.

[0164] The results are as follows Figure 4-6 As shown, Figure 4It is the garbage height before and after the garbage is fed into the pilot process equipment of Example 2 within one day (the horizontal axes 1-8 in the figure are different sampling points for collecting effluent. 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 dinner garbage feeding; 8: after dinner garbage feeding); Figure 5 It is the reduction rate of the height of the kitchen waste after being degraded by the composite bacteria of the present invention for one night (14 hours) (the horizontal axes 1-6 in the figure are the time points of the comparison of the height of the waste respectively); Figure 6 It is the COD removal rate of the effluent from the liquefied food waste after one night (14h) of degradation by composite bacteria (the horizontal axes 1-6 in the figure are COD removal rate versus time, respectively).

[0165] In summary, the composite bacteria can effectively degrade liquefied food waste.

[0166] It can be seen from the above embodiments that the present invention provides a composite bacteria for degrading liquefied kitchen waste and its application, wherein the composite bacteria includes 11 kinds of bacteria, namely, Pseudochrobactrum insaccharolyticum, Alcaligenes faecalis, Bacillus amylolyticus, Serratia marcescens, Sphingobacterium water, Bacillus subtilis, Achromobacter xylosoxidans, Bacillus cereus, Sphingobacterium tobacco soil, Stenotrophomonas maltophilia, Sphingobacterium humi, which are Pseudochrobactrumasaccharolyticum, Alcaligenesfaecalis, Paenibacillus amylolyticus, Serratiamarcescens, Sphingobacterium mizutaii, Bacillus subtilis, Achromobacterxylosoxidans, Bacillus cereus, Sphingobacterium tabacisoli, Stenotrophomonasmaltophilia, and Sphingobacterium humi in Latin. The seed numbers are CGMCCNO.27442, CGMCCNO.27443, CGMCCNO.27445, CGMCCNO.27448, CGMCCNO.27451, CGMCCNO.27452, CGMCCNO.27444, CGMCCNO.27446, CGMCCNO.27447, CGMCCNO.27449 and CGMCCNO.27450 respectively. The composite bacteria of the present invention are used to treat kitchen waste. In actual pilot applications, the composite bacteria can still achieve a good and continuous degradation effect on kitchen waste, and the waste reduction rate can reach 99.78% within one week. The culture conditions of the composite bacteria are easy to realize and control, and the composite bacteria can effectively degrade and liquefy kitchen waste, which is of great significance for the efficient treatment of kitchen waste.

[0167] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

Claims

1. A bacterial composition, which consists of the following eleven strains 1) to 11): 1) Pseudochrobactrum asaccharolyticum HL-1, whose registration number in the General Microbiological Center of China Culture Collection Administration is CGMCC No.27442; 2) Alcaligenes faecalis HL-2, whose registration number in the General Microbiological Center of China Culture Collection Administration is CGMCC No.27443; 3) Achromobacter xylosoxidans HL-3, whose registration number in the General Microbiological Center of China National Microbiological Culture Collection is CGMCC No.27444; 4) Paenibacillus amylolyticus HL-4, whose registration number in the General Microbiological Center of China Culture Collection Administration is CGMCC No.27445; 5) Bacillus cereus HL-14, whose registration number in the General Microbiological Center of China Culture Collection Administration is CGMCC No.27446; 6) Sphingobacterium tabacisoli HL-15, whose registration number in the General Microbiological Center of China Microbiological Culture Collection is CGMCC No.27447; 7) Serratia marcescens HL-18, whose registration number in the General Microbiological Center of China Culture Collection Administration is CGMCC No.27448; 8) Stenotrophomonas maltophilia HL-20, whose registration number in the General Microbiological Center of China Culture Collection Administration is CGMCC No.27449; 9) Sphingobacterium humi HL-25, whose registration number in the General Microbiological Center of China Microbiological Culture Collection is CGMCC No.27450; 10) Sphingobacterium mizutaii HL-26, whose registration number in the General Microbiological Center of China Microbiological Culture Collection is CGMCC No.27451; 11) Bacillus subtilis ( Bacillus subtilis ) HL-27, whose registration number in the General Microbiological Center of China Microbiological Culture Collection is CGMCC No.27452; Among them, the biomass ratio of the eleven strains is 1:1:1:1:1:1:1:1:1:1:1:

1.

2. The use of the bacterial composition according to claim 1 in the liquefaction and degradation of food waste, characterized in that: The bacterial composition was inoculated into a mixed solution of food and kitchen waste, and the initial biomass of each strain was 0.01-0.03 g / L.

3. A bacterial agent for liquefying and degrading kitchen waste, characterized in that: The active ingredient of the bacterial agent is the bacterial composition described in claim 1.

4. The bacterial agent according to claim 3, characterized in that The bacterial agent is formed by mixing single-bacteria enriched liquids of eleven strains in the bacterial composition described in claim 1, wherein the single-bacteria enriched liquid is a bacterial liquid obtained by inoculating the strains into an enriched culture liquid, wherein each liter of the enriched culture liquid comprises 3.0 g of beef extract, 10.0 g of peptone, and 5.0 g of NaCl, and the pH of the enriched culture liquid is 7.2-7.

4.

5. A method for liquefying and degrading kitchen waste, characterized in that: The bacterial composition according to claim 1 or the bacterial agent according to any one of claims 3-4 is inoculated into a mixed liquid of food waste; after inoculation into the mixed liquid of food waste, the initial biomass of each strain in the bacterial composition is 0.01-0.03 g / L, and the total biomass in the mixed liquid of food waste is 0.11-0.33 g / L.

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