Kitchen garbage rapid biodegradation complex microbial inoculant and preparation and application thereof

Abstract

The invention belongs to the field of kitchen waste resourceful treatment and microorganisms, and relates to a kitchen waste rapid biodegradation composite degradation bacterial agent as well as a preparation method and application of the degradation bacterial agent. The kitchen waste microbial complex microbial inoculant disclosed by the invention is a mixed microbial inoculant of bacillus and comprises lactobacillus paracasei FRG with the preservation number of CGMCC (China General Microbiological Culture Collection Center) 1.12731, bacillus subtilis K14 with the preservation number of CGMCC 1.14985, bacillus safensis SYB with the preservation number of CICC 21743, bacillus megaterium JYB with the preservation number of CICC 22681 and bacillus thiamine lyticum SSYB with the preservation number of DSM 10154. The kitchen waste microbial compound degradation bacterial agent can effectively degrade starch, protein, cellulose, grease and other characteristic components in kitchen waste, promote reduction of the kitchen waste and promote rapid decomposition of the kitchen waste.

Description

Technical Field

[0001] The invention belongs to the field of kitchen waste resource processing and microorganisms, and relates to a kitchen waste rapid biodegradable composite bacterial agent and a preparation method and application thereof. Background Art

[0002] Kitchen waste refers to organic waste generated in daily activities such as homes and restaurants, mainly including vegetable roots and leaves, fruit peels and cores, leftovers, bones and internal organs, and other organic solid waste. Aerobic fermentation of kitchen waste into organic fertilizer is one of the main ways to utilize it as a resource, but due to its high content of wood cellulose, protein, and oil, it is difficult to be naturally degraded, and to a certain extent, it leads to a longer fermentation cycle. If it is not handled properly and the fertilizer does not meet the standards, toxic substances will accumulate in the plant body and even enter the human body through the food chain, endangering human health.

[0003] Kitchen waste has complex components, high water content, high organic matter and salt content. The currently commonly used single strain and single function microbial agents still have limitations in the degradation and promotion of kitchen waste. Therefore, it is of great research significance to prepare microbial composite agents that are tolerant to oil and salt and have high-efficiency degradation functions for aerobic fermentation of kitchen waste, promote the resource utilization of kitchen waste, and reduce secondary pollution. Summary of the invention

[0004] The object of the present invention is to provide a kitchen waste microbial composite degradation agent and a preparation method and application thereof, so as to solve the problems of incomplete aerobic fermentation of kitchen waste and substandard organic fertilizer.

[0005] The invention firstly provides a kitchen waste microbial composite degradation agent, which comprises: Lactobacillus paracasei FRG (Lactobacillus paracasei) with a preservation number of CGMCC1.12731, Bacillus subtilis K14 (Bacillus subtilis) with a preservation number of CGMCC 1.14985, Bacillus safensis SYB (Bacillus safensis) with a preservation number of CICC 21743, Bacillus megaterium JYB (Bacillus megaterium) with a preservation number of CICC 22681 and Aneurinibacillus thermoaerophilus SSYB (Aneurinibacillus thermoaerophilus) with a preservation number of DSM 10154.

[0006] Specifically, Lactobacillus paracasei FRG, Bacillus subtilis K14, Bacillus saffron SYB, Bacillus megaterium JYB, and thermophilic aerobic thiamine-lytic Bacillus SSYB are mixed in a ratio of 4-6:3-5:2-4:4-6:2-4, preferably in a ratio of 5:4:3:5:3.

[0007] The present invention provides a method for preparing the kitchen waste microbial composite degradation agent, which comprises the following steps:

[0008] The activated strains were inoculated into liquid culture medium for activation, preferably until OD 600 The value is 0.8-1.2, and the number of colonies reaches 10 11 ~10 13 CFU / L;

[0009] The cultured Lactobacillus paracasei FRG, Bacillus subtilis K14, Bacillus sabdariffa SYB, Bacillus megaterium JYB and thermophilic aerobic thiamine-lytic Bacillus SSYB are mixed according to the ratio of the final colony count to form the kitchen waste microbial composite degradation agent.

[0010] Specifically, the activated strains are inoculated into nutrient gravy NA liquid culture medium or MRS liquid culture medium for culture; more specifically, the Lactobacillus paracasei FRG is inoculated and activated in the MRS liquid culture medium; the Bacillus subtilis K14, Bacillus saffron SYB, Bacillus megaterium JYB, and thermophilic aerobic thiamine-lytic Bacillus SSYB are inoculated and activated in the nutrient gravy NA liquid culture medium.

[0011] In a preferred embodiment, the culture conditions are as follows: the Lactobacillus paracasei FRG is cultured in a constant temperature shaking incubator at 28-32°C and 120-180 rpm / min for 24-48 hours; the Bacillus subtilis K14, Bacillus SYB, and Bacillus megaterium JYB are cultured in a constant temperature shaking incubator at 28-32°C and 120-180 rpm / min for 24-48 hours; the thermophilic aerobic thiamine-lytic Bacillus SSYB is cultured in a constant temperature shaking incubator at 50-60°C and 120-180 rpm / min for 24-48 hours.

[0012] More preferably, the culture conditions are as follows: the Lactobacillus paracasei FRG is cultured in a constant temperature shaking incubator at 30°C and 150rpm / min for 36 hours; the Bacillus subtilis K14, Bacillus SYB, and Bacillus megaterium JYB are cultured in a constant temperature shaking incubator at 30°C and 150rpm / min for 36 hours; the thermophilic aerobic thiamine-lytic Bacillus SSYB is cultured in a constant temperature shaking incubator at 55°C and 150rpm / min for 36 hours.

[0013] The present invention provides the application of the kitchen waste microbial composite degradation agent to resource treatment of kitchen waste, degradation of characteristic component protein, degradation of characteristic component oil, degradation of characteristic component cellulose and / or degradation of characteristic component starch.

[0014] The present invention further provides a method for resource processing of kitchen waste, which comprises the step of mixing the kitchen waste microbial composite deodorizing agent with the kitchen waste for aerobic fermentation.

[0015] Specifically, the kitchen waste is picked and minced, and bran is added (the weight ratio of kitchen waste to bran is 20-30:1) and mixed thoroughly, and the composite bacterial agent is introduced into the kitchen waste bran mixture and mixed evenly for solid fermentation; the resource recovery treatment refers to the degradation of the characteristic components of kitchen waste, such as protein, oil, cellulose and / or starch.

[0016] In a specific embodiment, the solid fermentation conditions are as follows: the fermentation temperature is controlled to rise from 25°C to 55°C on the 1st to 2nd day, the temperature is controlled to remain at 55°C on the 3rd to 8th day, the temperature is controlled to drop from 55°C to 25°C on the 9th to 12th day, and the compost is turned (stirred up and down with a glass rod) once a day.

[0017] The present invention selects strains that can efficiently degrade characteristic components of kitchen waste (protein, oil, cellulose and starch), mixes them in proportion and optimizes them to form the microbial composite degradation agent, which greatly improves the ability of a single strain to degrade characteristic substances of kitchen waste. At the same time, the antagonistic effect results of each strain in the microbial composite degradation agent show that there is no antagonism between strains, and the strains can achieve dominant growth in the aerobic fermentation environment of kitchen waste, thereby achieving the effect of promoting the reduction of kitchen waste and the degradation of characteristic components of kitchen waste.

[0018] The kitchen waste microbial composite degradation agent of the present invention has the following beneficial effects:

[0019] (1) In order to solve the problem of long aerobic fermentation cycle of kitchen waste, the present invention analyzes the dominant bacterial genera in the entire aerobic fermentation process of kitchen waste, screens out strains with efficient degradation function and optimizes the combination to form a microbial composite degradation agent, thereby providing a highly efficient, low-cost, and secondary pollution-free microbial composite degradation agent. In addition, since the microorganisms can survive for a long time in the aerobic fermentation environment of kitchen waste, the effect of promoting the reduction of kitchen waste and promoting the degradation of characteristic components of kitchen waste can be achieved.

[0020] (2) The present invention screened 1 strain of Lactobacillus and 4 strains of Bacillus, which have the ability to efficiently degrade characteristic components (protein, oil, cellulose and starch) in the aerobic fermentation process of kitchen waste. The selected strains were mixed in proportion and compounded and optimized to form a unique microbial composite degradation agent of the present invention. Compared with a single strain, the reduction effect of kitchen waste was effectively improved.

[0021] (3) The microbial composite degradation agent of the present invention does not produce secondary pollution and has low cost. The microbial composite degradation agent is composed of 1 strain of lactobacillus and 4 strains of bacillus. Most of the current microbial agents are single strains. The composite degradation agent of the present invention has a significantly higher reduction effect than single strains and is also higher than the existing microbial agents on the market. The microbial composite degradation agent of the present invention can survive for a long time during the aerobic fermentation of kitchen waste, and can achieve the effect of promoting the reduction of kitchen waste and the degradation of characteristic components of kitchen waste. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the abundance map of microbial communities after 45 days of aerobic fermentation of kitchen waste.

[0023] Figure 2 This is the plate colony morphology of the 5 strains screened in the present invention.

[0024] Figure 3 The figure shows the salt tolerance of the five strains screened by the present invention.

[0025] Figure 4 The present invention provides a microbial solid composite degradation agent.

[0026] Figure 5 The present invention shows the degradation effect of the microbial solid composite degradation agent on protein.

[0027] Figure 6 The invention discloses the degradation effect of the microbial solid composite degradation agent on starch.

[0028] Figure 7 The figure shows the degradation effects of the five strains screened by the present invention on oil.

[0029] Figure 8The present invention shows the degradation effect of the microbial solid composite degradation agent on oil and fat.

[0030] Fig. 9 The present invention shows the degradation effect of the microbial solid composite degradation agent on cellulose.

[0031] Fig.10 The present invention provides statistics on the degradation effect of the microbial solid composite degradation agent prepared in the present invention on the characteristic components of kitchen waste.

[0032] Fig.11 The invention discloses the kitchen waste reduction effect of the microbial solid composite degradation agent prepared by the invention. DETAILED DESCRIPTION

[0033] The following further describes the content of the present invention in detail by combining the accompanying drawings and specific embodiments. The specific embodiments are intended to facilitate a better understanding of the present invention, but are not intended to limit the present invention in any way.

[0034] Example 1: Preparation of microbial solid composite agent

[0035] 1. Preparation of pure microbial culture solution

[0036] A 45-day aerobic fermentation experiment was conducted on kitchen waste, and the abundance map of the microbial community during the entire aerobic fermentation process was obtained, such as Figure 1 As shown in the figure, by analyzing the dominant bacterial genera in the entire aerobic fermentation process of kitchen waste, the phylum-level bacteria and genus-level bacteria with the highest relative abundance were obtained, and one lactobacillus and four bacillus strains with high-efficiency degradation function were screened out. They are: Lactobacillus paracasei FRG with a preservation number of CGMCC 1.12731, Bacillus subtilis K14 with a preservation number of CGMCC 1.14985, Bacillus safensis SYB with a preservation number of CICC21743, Bacillus megaterium JYB with a preservation number of CICC 22681, and Aneurinibacillus thermoaerophilus SSYB with a preservation number of DSM 10154.

[0037] The selected lactobacillus and four bacillus strains were inoculated and activated in MRS liquid medium and nutrient broth (NA) liquid medium, respectively. After three generations of subculture, the bacterial liquid of each single strain was taken and spread in MRS solid medium and nutrient broth (NA) solid medium for culture. After culture, a single colony of each strain was picked and cultured in MRS liquid medium and nutrient broth (NA) liquid medium, respectively, to obtain the pure bacterial liquid of each single strain.

[0038] 2. Culture medium

[0039] MRS medium was used for the culture of Lactobacillus paracasei FRG, nutrient broth (NA) medium was used for the culture of Bacillus subtilis K14, Bacillus safensisSYB, Bacillus megaterium JYB, and Aneurinibacillus thermoaerophilus SSYB, and lysing broth (LB) solid medium was used to test the antagonism between strains.

[0040] The formula of MRS liquid culture medium is as follows: 10.0 g / L tryptone, 10.0 g / L beef extract, 5.0 g / L yeast extract powder, 20.0 g / L glucose, 1.0 mL Tween-80, 2.0 g / L ammonium citrate, 5.0 g / L sodium acetate, 2.0 g / L dipotassium hydrogen phosphate, 0.1 g / L magnesium sulfate, 0.05 g / L manganese sulfate, and 1 L distilled water. 15.0 g / L agar should also be added to the MRS solid culture medium.

[0041] The formula of nutrient broth (NA) liquid culture medium is as follows: 10.0 g / L tryptone, 3.0 g / L beef extract, 0.005 g / L manganese sulfate, 5.0 g / L sodium chloride (NaCl), 1 L distilled water, and 15.0 g / L agar should also be added to the NA solid culture medium.

[0042] The formula of lysate broth (LB) solid culture medium is as follows: tryptone 10.0 g / L, sodium chloride (NaCl) 10.0 g / L, yeast extract powder 5.0 g / L, agar 15.0 g / L, and distilled water 1 L.

[0043] 3. Antagonism between strains

[0044] Pure bacterial suspensions of the above strains were cultured in pairs on lysing broth (LB) solid culture medium in a cross-line pattern to test the antagonism between the strains in the microbial composite agent. The results showed that there was no antagonism between the strains.

[0045] 4. Salt tolerance of each strain

[0046] NaCl with concentrations of 1%, 3%, 5% and 7% was added to NA liquid medium, MRS liquid medium and YPD liquid medium respectively, mixed evenly, sterilized at 121℃ for 15min to prepare each salt-containing liquid medium, and the bacterial concentration of each strain was measured after cultivation, and the tolerance to salt was judged by the size of the bacterial concentration. Figure 3 As shown in the figure, when the salt concentration was 1% and 3%, all five strains had good growth characteristics; when the salt concentration reached 5%, the growth rates of the two strains K14 and SSYB were low, and a certain inhibitory effect was produced, and neither of them reached the logarithmic growth phase; when the salt concentration reached 7%, only strain FRG could grow stably and reach the logarithmic growth phase. In summary, the five strains screened had good growth conditions when the salt concentration was below 5%.

[0047] 5. Cultivation of microbial suspension

[0048] Lactobacillus paracasei FRG was cultured in a constant temperature shaking incubator at 30°C and 150 rpm / min for 36 hours; Bacillus subtilis K14, Bacillus safensis SYB, and Bacillus megaterium JYB were cultured in a constant temperature shaking incubator at 30°C and 150 rpm / min for 36 hours; and Aneurinibacillus thermoaerophilus SSYB was cultured in a constant temperature shaking incubator at 55°C and 150 rpm / min for 36 hours to prepare an OD 600 =1.0 bacterial suspension.

[0049] After culturing under the above culture conditions, the colony count of Lactobacillus paracasei FRG can reach 6.58×10 11 CFU / L, the colony count of Bacillus subtilis K14 can reach 1.83×10 12CFU / L, the colony count of Bacillus safensis SYB can reach 1.70×10 12 CFU / L, the colony count of Bacillus megaterium JYB can reach 8.05×10 11 CFU / L, the colony count of Aneurinibacillus thermoaerophilus SSYB can reach 1.67×10 11 CFU / L.

[0050] 5. Preparation of composite bacterial agent

[0051] Lactobacillus paracasei FRG, Bacillus subtilis K14, Bacillus safensis SYB, Bacillus megaterium JYB and Aneurinibacillus thermoaerophilus SSYB are mixed in a ratio of 5:4:3:5:3 according to the final colony count, and strain protectants (10 g / L glycerol, 10 g / L skimmed milk powder, 10 g / L soluble starch) are added thereto. After mixing evenly, the mixture is pre-cooled at -80°C overnight, and then freeze-dried in a vacuum freeze dryer for 24 hours to obtain a microbial solid composite degradation agent.

[0052] Example 2: Degradation effect of microbial solid composite degradation agent on protein

[0053] Preparation of egg solid culture medium: Place egg white in a culture dish, then place it in a sterilizer, and sterilize it at 121°C for 20 min to obtain an egg solid culture medium. The degradation effect of the composite degradation bacteria on protein is determined by the egg solid culture medium. The prepared composite degradation bacteria is added to the egg solid culture medium, inverted and cultured at 37°C for 2 days, and the quality difference before and after the egg solid culture medium is used to evaluate the ability of the composite degradation bacteria to degrade protein substances.

[0054] The protein degradation effect of each single strain was verified according to Xu Jing's hydrolysis circle method. The formula of protein culture medium is as follows: Na2HPO4·7H2O 1.07g / L, casein 5.0g / L, KH2PO4 0.36g / L, agar 20.0g / L, distilled water 1L. According to the formula of protein culture medium, solid plate culture medium was prepared, 30 mL of the bacterial liquid of 5 strains was poured into a centrifuge tube, centrifuged at 8000 r / min for 8 min, and the supernatant was poured to obtain the bacterial mud of the strain. The bacterial mud was dipped into the protein solid culture medium with an inoculation loop and incubated for 24 h at the respective culture temperature, and then taken out and the size of the hydrolysis circle on the culture medium was observed. The degradation ability of the strain on protein substances was measured by the ratio of the hydrolysis circle diameter (D) to the colony diameter (d). As can be seen from the table below, the degradation abilities of the five strains for protein substances are, from large to small, respectively: Bacillus subtilis K14, Bacillus megaterium JYB, Bacillus safensis SYB, Lactobacillus paracasei FRG, while Aneurinibacillus thermoaerophilus SSYB did not produce obvious hydrolysis circles.

[0055]

[0056] like Figure 5 As shown, after 2 days of degradation before and after inoculation of the microbial composite degradation agent, the protein surface undergoes significant changes. In the solid medium of eggs treated with microorganisms, the surface turns yellow and has a mixed sour taste. Fig.11 It can be seen that the degradation effect of the composite degradation bacteria on protein is about 3 times that of the blank group CK. The average mass difference of the blank group CK after protein degradation is 0.570 g, and the average mass difference of the composite degradation bacteria after protein degradation is 1.515 g.

[0057] Example 3: Degradation effect of microbial solid composite degrading agent on starch

[0058] Preparation of rice solid culture medium: Put the cooked rice into a culture dish, then put it into a sterilizer, and sterilize it at 121℃ for 20 min to obtain the rice solid culture medium. The degradation effect of the composite degradation bacteria on starch is determined by the rice solid culture medium. The prepared composite degradation bacteria is added to the rice solid culture medium, and cultured inverted at 37℃ for 2 days. The quality difference before and after the rice solid culture medium is used to evaluate the ability of different composite bacteria to degrade protein substances.

[0059] The degradation effect of each single strain on starch was verified according to Xu Jing's hydrolysis circle method. The formula of starch culture medium is as follows: NaCl 5.0g / L, tryptone 5.0g / L, soluble starch 20.0g / L, agar 20.0g / L, distilled water 1L. According to the formula of starch culture medium, solid plate culture medium was prepared, 30 mL of the bacterial liquid of 5 strains was poured into a centrifuge tube, centrifuged at 8000 r / min for 8 min, and the supernatant was poured to obtain the bacterial mud of the strain. Dip the bacterial mud with an inoculation loop and apply it to the starch solid culture medium, and invert and culture it for 24 h at the respective culture temperature. After the culture is completed, the plate culture medium is taken out and iodine solution is dripped into the culture medium (the hydrolysis circle can be determined according to the characteristic that starch turns blue when it encounters iodine). The size of the hydrolysis circle on the culture medium is observed on the black desktop. The degradation ability of the strain on starch substances is measured by the ratio of the hydrolysis circle diameter (D) to the colony diameter (d). As can be seen from the table below, the degradation abilities of the five strains for starch substances are, from high to low, respectively: Bacillus subtilis K14, Bacillus megaterium JYB, Lactobacillus paracasei FRG, Bacillus safensis SYB and Aneurinibacillus thermoaerophilus SSYB, which did not produce obvious hydrolysis circles.

[0060]

[0061] Depend on Figure 6 As shown in the figure, after 2 days of degradation before and after inoculation of the microbial composite degradation agent, the surface of the rice changed significantly, from white to brown, and more water droplets condensed on the glass surface of the culture dish, and the rice became mushy with a slightly sour and smelly taste. Fig.11 It can be seen that the degradation effect of the composite degrading bacteria on protein is about 1.5 times that of the blank group CK. The average mass difference of the blank group CK after starch degradation is 1.2475 g, and the average mass difference of the composite degrading bacteria after starch degradation is 2.0275 g.

[0062] Example 4: Degradation effect of microbial solid composite degradation agent on oil and fat

[0063] Preparation of liquid culture medium for inorganic salts of oils and fats: The formula of liquid culture medium for inorganic salts of oils and fats is as follows: (NH4)2SO45.0g / L, KH2PO4 2.0g / L, MgSO4 0.024g / L, NaCl 2.0g / L, NaH2PO4 2.0g / L, soybean oil 10.0g / L, and distilled water 1L. Prepare liquid culture medium for inorganic salts of oils and fats with natural pH, put it in a sterilizing pot, and sterilize it at 121°C for 20 min to obtain liquid culture medium for inorganic salts of oils and fats. The degradation effect of the composite degrading bacterial agent on oils and fats is determined by the liquid culture medium for inorganic salts of oils and fats. Subsequently, the composite degrading bacterial agent is added to the liquid culture medium for inorganic salts of oils and fats and cultured for 7 days. The oils and fats in the fermented liquid after 7 days of culture are extracted by petroleum ether, shaken thoroughly and allowed to stand for 30 min, and the upper layer of liquid is taken and appropriately diluted to determine the OD 225 The value was calculated and the oil degradation rate was calculated based on the standard curve to calculate the concentration of residual oil.

[0064] The degradation effect of each single strain on oil was verified according to the degree of neutral red color change. The formula of oil culture medium is as follows: tryptone 10.0g / L, Tween-80 10.0g / L, NaCl 10.0g / L, magnesium sulfate 0.048g / L, agar 20.0g / L, distilled water 1L. According to the formula of oil culture medium, prepare solid plate culture medium, pour 30 mL of the bacterial liquid of 5 strains into a centrifuge tube, centrifuge at 8000 r / min for 8 min, pour off the supernatant, and obtain the strain mud. Use an inoculation loop to dip the mud into the oil solid culture medium and invert it for 24 hours at the respective culture temperature. After the culture is completed, take out the plate culture medium, add neutral red reagent and cover it for 15 minutes. By Figure 7 It can be seen that among the five strains, Bacillus subtilis K14, Bacillus safensis SYB, Bacillus megaterium JYB and Aneurinibacillus thermoaerophilus SSYB have stronger ability to degrade oil, while Lactobacillus paracasei FRG has weaker ability to degrade oil.

[0065] like Figure 8 As shown in the figure, after 7 days of degradation before and after the inoculation of the microbial composite degradation agent, it was found that the color of the culture medium added with the microbial composite degradation agent changed from transparent to milky white. Fig.11It can be seen that the degradation effect of the composite degradation bacteria on oil is about 10 times that of the blank group CK group. The degradation rate of the blank group CK on oil is 4.01%, and the degradation rate of the composite degradation bacteria on oil is 42.15%.

[0066] Example 5: Degradation effect of microbial solid composite degradation agent on cellulose

[0067] Preparation of Hutcheson's inorganic salt medium: The formula of Hutcheson's inorganic salt medium is as follows: KH2PO4 1.0g / L, NaNO3 2.5g / L, MgSO4 0.15g / L, NaCl 0.1g / L, CaCl2 0.1g / L, distilled water 1L. Prepare Hutcheson's inorganic salt medium, pH natural, put it in a sterilizer, sterilize it at 121℃ for 20 min to obtain Hutcheson's inorganic salt medium. The degradation effect of the composite degrading bacteria on cellulose is determined by Hutcheson's inorganic salt medium. Add the composite degrading bacteria to the culture medium for cultivation. Under the conditions of 37℃, 200r / min, constant temperature shaking culture for 7 days, take out the sample, place the residue in an 80℃ oven to dry and weigh, and use the improved Van Soest washing method to determine the sample weight loss rate to calculate the degradation rate of cellulose by the composite degrading bacteria.

[0068] The formula of cellulose culture medium is as follows: CMC-Na 6.25 g / L, NaCl 0.125 g / L, CaCl20.0125 g / L, KH2PO41.25 g / L, MgSO40.375 g / L, FeCl30.0125 g / L, agar 20.0 g / L, distilled water 1L. According to the formula of cellulose culture medium, solid plate culture medium was prepared, 30 mL of 15 strains of bacterial liquid was poured into a centrifuge tube, centrifuged at 8000 r / min for 8 min, and the supernatant was poured to obtain the strain mud. The bacterial mud was dipped into the starch solid culture medium with an inoculation loop and inverted for 24 h at the respective culture temperature. After the culture was completed, the plate culture medium was taken out, 0.1% Congo red dye was added to it, and the treatment was covered for 15 min, followed by a drop of 1 mol / L NaCl solution, and the treatment was covered for 15 min. The ratio of the hydrolysis zone diameter (D) to the colony diameter (d) was used to measure the strain's ability to degrade cellulose. The cellulose degradation effect of each single strain was verified according to Xu Jing's hydrolysis zone method. As shown in the table below, the thermophilic aeroaerophilic thiamine-degrading Bacillus SSYB (Aneurinibacillusthermoaerophilus SSYB) was the most obvious one among the five strains, while the other four strains did not produce obvious hydrolysis zones.

[0069]

[0070] Depend on Fig. 9 As shown in the figure, after 7 days of degradation before and after the inoculation of the microbial composite agent, the corn stalk powder in the culture medium dissolved in the culture medium, and the culture medium changed from transparent to turbid. Fig.11 It can be seen that the degradation effect of the composite degrading bacteria on cellulose is about 6 times that of the blank group CK group. The cellulose degradation rate of the blank group CK is 6.72%, and the cellulose degradation rate of the composite degrading bacteria is 37.58%.

[0071] Example 6: Effect of microbial solid composite degradation agent on the reduction of kitchen waste

[0072] The kitchen waste from the canteen was picked and minced, and 10g of bran was added to every 40g of kitchen waste for thorough mixing. The composite bacterial agent was added to the kitchen waste bran mixture and mixed evenly. The mixture was placed in a 150mL conical flask for a 12-day aerobic fermentation test. The temperature of the incubator was adjusted to simulate the temperature change during aerobic fermentation: the fermentation temperature was controlled from 25℃ to 55℃ on the 1st to 2nd day, the temperature was controlled to be maintained at 55℃ on the 3rd to 8th day, and the temperature was controlled to be reduced from 55℃ to 25℃ on the 9th to 12th day. The pile was turned over once a day (stirred up and down with a glass rod), and samples were taken every 2 days and weighed and recorded. The weight loss results of kitchen waste before and after aerobic fermentation represent the degradation rate of kitchen waste by each single strain and composite degradation bacterial agent.

[0073] Depend on Fig.11It can be seen that the degradation rate of kitchen waste by the blank group CK is only 17.14%; the degradation rate of kitchen waste by the composite degradation bacteria is 71.39%, which is 316.51% higher than that of the blank group CK; the degradation rate of kitchen waste by Lactobacillus paracasei FRG is 17.42%, which is 1.63% higher than that of the blank group CK; the degradation rate of kitchen waste by Bacillus subtilis K14 is 42.06%, which is 145.39% higher than that of the blank group CK; the degradation rate of kitchen waste by Bacillus safensis SYB is 40.75%, which is 137.75% higher than that of the blank group CK; the degradation rate of kitchen waste by Bacillus megaterium JYB is 41.8%, which is 10.2% higher than that of the blank group CK; the degradation rate of kitchen waste by Bacillus sphaeroides is 1.3%, which is 2.2% higher than that of the blank group CK; the degradation rate of kitchen waste by Bacillus sphaeroides is 1.2%, which is 2.3% higher than that of the blank group CK; the degradation rate of kitchen waste by Bacillus sphaeroides is 1.1 ...3% higher than that of the blank group CK; the degradation rate of kitchen waste by Bacillus sphaeroides is 1.1%, which is 2.3% higher than that of the blank group CK. The degradation rate of kitchen waste by Bacillus thermoaerophilus SSYB was 39.52%, which was 130.57% higher than that of the blank group CK; the degradation rate of kitchen waste by Aneurinibacillus thermoaerophilus SSYB was 61.89%, which was 261.09% higher than that of the blank group CK. It can be seen that the degradation rate of kitchen waste is low without the addition of composite bacterial agents or single strains, indicating that it is difficult to degrade in a short time, while the addition of the prepared microbial composite bacterial agent has greatly improved the degradation rate of kitchen waste, and the degradation effect of the microbial composite degradation agent on kitchen waste is better than that of each single strain, and the degradation effect of the single strain on kitchen waste is better than that of the blank group CK.

Claims

1. A kitchen waste microbial composite degradation agent, characterized in that: The degradation bacterial agent is a mixed bacterial agent of lactobacillus and bacillus, including: Lactobacillus paracasei FRG (Lactobacillus paracasei) with a preservation number of CGMCC 1.12731, Bacillus subtilis K14 (Bacillus subtilis) with a preservation number of CGMCC 1.14985, Bacillus safensis SYB (Bacillus safensis) with a preservation number of CICC 21743, Bacillus megaterium JYB (Bacillus megaterium) with a preservation number of CICC 22681, and Aneurinibacillus thermoaerophilus SSYB (Aneurinibacillus thermoaerophilus) with a preservation number of DSM 10154.

2. The kitchen waste microbial composite degradation agent according to claim 1, characterized in that: Lactobacillus paracasei FRG, Bacillus subtilis K14, Bacillus saffron SYB, Bacillus megaterium JYB, and thermophilic aerobic thiamine-lytic Bacillus SSYB are mixed in a ratio of 4-6:3-5:2-4:4-6:2-4, preferably in a ratio of 5:4:3:5:

3.

3. The method for preparing the kitchen waste microbial composite degradation agent according to claim 1 or 2, characterized in that: The following steps are involved: The activated strains were inoculated into liquid culture medium for activation, preferably until OD 600 The value is 0.8-1.2, and the number of colonies reaches 10 11 ~10 13 CFU / L; The cultured Lactobacillus paracasei FRG, Bacillus subtilis K14, Bacillus sabdariffa SYB, Bacillus megaterium JYB and thermophilic aerobic thiamine-lytic Bacillus SSYB are mixed according to the ratio of the final colony count to form the kitchen waste microbial composite degradation agent.

4. The method for preparing the kitchen waste microbial composite degradation agent according to claim 3, characterized in that: The activated strains are inoculated into nutrient broth NA liquid culture medium or MRS liquid culture medium for culture. The Lactobacillus paracasei FRG is inoculated and activated in the MRS liquid culture medium; the Bacillus subtilis K14, Bacillus saffron SYB, Bacillus megaterium JYB, and thermophilic aerothiamine-lytic Bacillus SSYB are inoculated and activated in the nutrient broth NA liquid culture medium.

5. The method for preparing the kitchen waste microbial composite degradation agent according to claim 3, characterized in that: The culture conditions are as follows: the Lactobacillus paracasei FRG is cultured in a constant temperature shaking incubator at 28-32°C and 120-180 rpm / min for 24-48 hours; the Bacillus subtilis K14, Bacillus SYB, and Bacillus megaterium JYB are cultured in a constant temperature shaking incubator at 28-32°C and 120-180 rpm / min for 24-48 hours; and the thermophilic aerobic thiamine-lytic Bacillus SSYB is cultured in a constant temperature shaking incubator at 50-60°C and 120-180 rpm / min for 24-48 hours.

6. The method for preparing the kitchen waste microbial composite degradation agent according to claim 5, characterized in that: The culture conditions are as follows: the Lactobacillus paracasei FRG is cultured in a constant temperature shaking incubator at 30°C and 150rpm / min for 36 hours; the Bacillus subtilis K14, Bacillus SYB, and Bacillus megaterium JYB are cultured in a constant temperature shaking incubator at 30°C and 150rpm / min for 36 hours; and the thermophilic aerobic thiamine-solubilizing Bacillus SSYB is cultured in a constant temperature shaking incubator at 55°C and 150rpm / min for 36 hours.

7. Application of the kitchen waste microbial composite degrading agent as claimed in claim 1 to resource treatment of kitchen waste, degradation of characteristic component protein, degradation of characteristic component fat, degradation of characteristic component cellulose and / or degradation of characteristic component starch.

8. A method for recycling kitchen waste, comprising the following steps: mixing the kitchen waste microbial composite deodorizing agent as claimed in claim 1 or 2 with kitchen waste for aerobic fermentation.

9. The method according to claim 8, characterized in that The kitchen waste is picked and minced, and bran is added (the weight ratio of kitchen waste to bran is 20-30:1), mixed thoroughly and put into a triangular flask, and the composite bacterial agent is introduced into the kitchen waste bran mixture and mixed evenly for solid fermentation; the resource recovery treatment refers to the degradation of the characteristic components of kitchen waste, such as protein, oil, cellulose and / or starch.

10. The method according to claim 9, characterized in that The solid fermentation conditions are as follows: the fermentation temperature is controlled to rise from 25°C to 55°C on the 1st to 2nd day, the temperature is controlled to remain at 55°C on the 3rd to 8th day, the temperature is controlled to drop from 55°C to 25°C on the 9th to 12th day, and the compost is turned (stirred up and down with a glass rod) once a day.

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