CN104531550A - High-temperature kitchen waste degradation microbial agent and preparing and using method thereof - Google Patents
High-temperature kitchen waste degradation microbial agent and preparing and using method thereof Download PDFInfo
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- 230000015556 catabolic process Effects 0.000 title claims abstract description 21
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 14
- 230000000813 microbial effect Effects 0.000 title claims abstract description 10
- 239000010806 kitchen waste Substances 0.000 title abstract 8
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 200
- 241000894006 Bacteria Species 0.000 claims abstract description 97
- 241000193385 Geobacillus stearothermophilus Species 0.000 claims abstract description 67
- 239000000843 powder Substances 0.000 claims abstract description 65
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 48
- 238000000855 fermentation Methods 0.000 claims abstract description 35
- 230000004151 fermentation Effects 0.000 claims abstract description 35
- 230000001580 bacterial effect Effects 0.000 claims abstract description 18
- 230000004913 activation Effects 0.000 claims description 126
- 239000007788 liquid Substances 0.000 claims description 110
- 229910052736 halogen Inorganic materials 0.000 claims description 66
- 150000002367 halogens Chemical class 0.000 claims description 66
- 239000010794 food waste Substances 0.000 claims description 54
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical group O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 49
- 239000001888 Peptone Substances 0.000 claims description 48
- 108010080698 Peptones Proteins 0.000 claims description 48
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 48
- 239000012153 distilled water Substances 0.000 claims description 48
- 235000019319 peptone Nutrition 0.000 claims description 48
- 239000002904 solvent Substances 0.000 claims description 48
- 238000002360 preparation method Methods 0.000 claims description 40
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 36
- 229940041514 candida albicans extract Drugs 0.000 claims description 36
- 239000008103 glucose Substances 0.000 claims description 36
- 238000011081 inoculation Methods 0.000 claims description 36
- 239000012138 yeast extract Substances 0.000 claims description 36
- 238000012258 culturing Methods 0.000 claims description 30
- 239000002054 inoculum Substances 0.000 claims description 26
- 238000012360 testing method Methods 0.000 claims description 25
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 24
- 229930195725 Mannitol Natural products 0.000 claims description 24
- 239000000594 mannitol Substances 0.000 claims description 24
- 235000010355 mannitol Nutrition 0.000 claims description 24
- 238000004519 manufacturing process Methods 0.000 claims description 24
- 229920002472 Starch Polymers 0.000 claims description 19
- 239000008107 starch Substances 0.000 claims description 19
- 235000019698 starch Nutrition 0.000 claims description 19
- ZPLCXHWYPWVJDL-UHFFFAOYSA-N 4-[(4-hydroxyphenyl)methyl]-1,3-oxazolidin-2-one Chemical compound C1=CC(O)=CC=C1CC1NC(=O)OC1 ZPLCXHWYPWVJDL-UHFFFAOYSA-N 0.000 claims description 12
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 12
- 229920000053 polysorbate 80 Polymers 0.000 claims description 12
- 241000196324 Embryophyta Species 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 230000003213 activating effect Effects 0.000 claims description 6
- 238000010564 aerobic fermentation Methods 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 3
- 238000010521 absorption reaction Methods 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 claims description 2
- 244000052616 bacterial pathogen Species 0.000 abstract description 5
- 230000000593 degrading effect Effects 0.000 abstract description 4
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 3
- 239000002131 composite material Substances 0.000 abstract description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract 2
- 241001626813 Anoxybacillus Species 0.000 abstract 1
- 241000588724 Escherichia coli Species 0.000 abstract 1
- 230000009286 beneficial effect Effects 0.000 abstract 1
- 229910052757 nitrogen Inorganic materials 0.000 abstract 1
- 230000009467 reduction Effects 0.000 description 22
- 239000001913 cellulose Substances 0.000 description 7
- 229920002678 cellulose Polymers 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 239000002068 microbial inoculum Substances 0.000 description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 4
- 230000009849 deactivation Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 241000256113 Culicidae Species 0.000 description 2
- 241000255925 Diptera Species 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- MKYBYDHXWVHEJW-UHFFFAOYSA-N N-[1-oxo-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propan-2-yl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(C(C)NC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 MKYBYDHXWVHEJW-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
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Abstract
The invention discloses a high-temperature kitchen waste degradation microbial agent and a preparing and using method thereof. A microbial strain comprises, by weight, 15-30 parts of nuby land anoxybacillus, 10-25 parts of liquefaction thermophilic bacillus, 20-40 parts of hot nitrogen bacillus and 25-50 parts of geobacillus stearothermophilus. Compared with the prior art, the high-temperature kitchen waste degradation microbial agent has the beneficial effects that as for kitchen waste with different integrants, reasonable matching on single bacterium powder can be carried out, and single bacterial strains are cooperated for degrading the kitchen waste in a collaborative mode; the number of effective viable bacteria in the single bacterium powder in the composite microbial agent is large, and when the microbial agent is used for degrading the kitchen waste, only a small number of microbial agents is needed for sufficiently and rapidly degrading the kitchen waste; components in the microbial agent are high-temperature cultures, the optimum fermentation temperature ranges from 55 DEG C to 65 DEG C, pathogenic bacteria such as escherichia coli and staphylococcus aureus in the kitchen waste can be effectively inactivated in the fermentation process, and the biosecurity is high.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of high temperature degradation changing food waste microbiobacterial agent and preparation thereof, using method.
Background technology
Changing food waste has the features such as high oil, high salt, high protein content, be easy to grow mosquitos and flies, rot if dealt with improperly, and distribute stench, grow mosquitos and flies, some changing food wastes are even used for refining " sewer oil ", feeding " swill pig " etc. by lawless person, cause serious impact to environment and health of people.
At present, the biological treatment of changing food waste, mainly contains mesophilic microorganism facture and high temperature microbe facture.
The leavening temperature of mesophilic microorganism facture is generally 25 ~ 40 DEG C, and the leavening temperature of pyroprocess can reach more than 50 DEG C.
Due to most of pathogenic bacterium, the optimum growth temperature of such as streptococcus aureus, intestinal bacteria etc. is about 35 DEG C, by mesophilic microorganism facture, fermentative processing is carried out to changing food waste, the pathogenic bacterium of non-deactivation in its output object, may be there are, there is potential safety hazard to a certain degree.
If carry out fermentative processing by high temperature microbe facture to changing food waste, then while fermenting process, deactivation can be carried out to the pathogenic bacterium existed in changing food waste.And the optimum temperuture of general enzyme is 50 ~ 60 DEG C, adopts the fermentation of high temperature microbe facture, the enzyme in bacterial classification can be made to play maximum degradation rate, shorten fermentation time, reduce fermentation energy consumption.
Summary of the invention
For the above-mentioned technical problem existed in prior art, the present invention proposes a kind of high temperature degradation changing food waste microbiobacterial agent, this microbiobacterial agent, as the microbial inoculum used in high temperature microbe facture, effectively can be degraded to changing food waste.
To achieve these goals, the present invention adopts following technical scheme:
A kind of high temperature degradation changing food waste microbiobacterial agent, described microbial strains is composited by the strain raw material of following mass fraction: exert than halogen ground anaerobic genus bacillus 15-30 part, thermophilic liquefaction genus bacillus 10-25 part, hot denitrogenation ground bacillus 20-40 part, Geobacillus stearothermophilus 25-50 part.
The invention allows for a kind of preparation method of high temperature degradation changing food waste microbiobacterial agent, it adopts following technical scheme:
A preparation method for high temperature degradation changing food waste microbiobacterial agent, comprises following preparation process:
S1, will exert and carry out individual plant cultivation respectively than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus, then obtained individual plant bacterium colony is carried out activation culture according in the inoculum size access liquid nutrient medium of 10-20%, then volume ratio is housed is ferment in the fermentation equipment of 60-75% substratum by the bacterial classification access after activation;
Adsorbed by gained fermented liquid carrier, obtained living bacteria count is 5 × 10
9individual/gram exert bacterium powder more single than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus; Absorption carrier is diatomite;
S2, obtain and exert than halogen ground anaerobic genus bacillus 15-30 part, thermophilic liquefaction genus bacillus 10-25 part, hot denitrogenation ground bacillus 20-40 part, Geobacillus stearothermophilus 25-50 part, the mass fraction of single for above-mentioned each bacterial strain bacterium powder is fully mixed to get microbiobacterial agent.
In above-mentioned steps s1, exert and than the single bacterium powder preparation process of halogen ground anaerobic genus bacillus be:
S111, exerting on the test tube slant after purified be seeded to than halogen ground anaerobic genus bacillus volume ratio is housed is in the triangular flask of liquid activation medium of 10-20%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, yeast extract paste 3-5g/L, Zulkovsky starch 15-35g/L, sodium-chlor 4-6g/L, and the pH value of liquid activation medium is 7.0-7.2; Inoculation exert than halogen ground anaerobic genus bacillus liquid activation medium temperature 55-65 DEG C, under shaking speed is 140-160r/min, cultivate 18-24h;
S112, will activate exerting to be seeded to than halogen ground anaerobic genus bacillus and being equipped with in fermentor tank that volume ratio is the fermention medium of 60-75% of gained through step s111, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, yeast extract paste 3-5g/L, Zulkovsky starch 15-35g/L, sodium-chlor 4-6g/L, and the pH value of fermention medium is 7.0-7.2; Inoculation exert than halogen ground anaerobic genus bacillus fermention medium temperature 55-65 DEG C, under mixing speed is 200-220r/min, enlarged culturing 20-30h, until in fermentor tank to exert than halogen ground anaerobic genus bacillus spore production rate be more than 80%, exerting after fermentation is adsorbed, until living bacteria count reaches 5 × 10 than halogen ground anaerobic genus bacillus carrier
9individual/gram, the single bacterium powder than halogen ground anaerobic genus bacillus must be exerted.
In above-mentioned steps s1, the single bacterium powder preparation process of thermophilic liquefaction genus bacillus is:
S121, the thermophilic liquefaction genus bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10-20%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, tween 80 20-35ml/L, N.F,USP MANNITOL 15-25g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of liquid activation medium is 7.0-7.2; The liquid activation medium inoculating thermophilic liquefaction genus bacillus temperature 55-60 DEG C, under shaking speed 140-160r/min, cultivate 18-24h;
S122, to be seeded to by the thermophilic liquefaction genus bacillus activating gained to be equipped with in fermentor tank that volume ratio is the fermention medium of 60-75% through step s121, inoculum size is 2-4%; The solvent of fermention medium is distilled water, solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, tween 80 20-35ml/L, N.F,USP MANNITOL 15-25g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of fermention medium is 7.0-7.2; The fermention medium inoculating thermophilic liquefaction genus bacillus temperature 55-62 DEG C, under mixing speed is 200-220r/min, enlarged culturing 24-36h, until the thermophilic liquefaction genus bacillus spore production rate in fermentor tank is more than 80%, thermophilic liquefaction genus bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of thermophilic liquefaction genus bacillus.
In above-mentioned steps s1, the single bacterium powder preparation process of hot denitrogenation ground bacillus is:
S131, the denitrogenation ground bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10-25%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, N.F,USP MANNITOL 15-25g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of liquid activation medium is 7.0-7.2; The liquid activation medium of inoculation denitrogenation ground bacillus temperature 55-65 DEG C, under shaking speed is 140-160r/min, cultivate 18-24h;
S132, be seeded to be equipped with activating the denitrogenation ground bacillus of gained through step s131 in fermentor tank that volume ratio is the fermention medium of 60-75%, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, N.F,USP MANNITOL 15-25g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of fermention medium is 7.0-7.2; The fermention medium of inoculation denitrogenation ground bacillus temperature 55-65 DEG C, under mixing speed is 200-220r/min, enlarged culturing 24-40h, until the denitrogenation ground bacillus spore production rate in fermentor tank is more than 80%, denitrogenation ground bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of denitrogenation ground bacillus.
In above-mentioned steps s1, the single bacterium powder preparation process of Geobacillus stearothermophilus is:
S141, the Geobacillus stearothermophilus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10-20%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 8-12g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, Bromothymol blue solution trace, and the pH value of liquid activation medium is 7.0-7.2; The liquid activation medium of inoculation Geobacillus stearothermophilus temperature 55-65 DEG C, under shaking speed is 140-160r/min, cultivate 18-24h;
S142, be seeded to be equipped with activating the Geobacillus stearothermophilus of gained through step s141 in fermentor tank that volume ratio is the fermention medium of 60-75%, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, Bromothymol blue solution trace, and the pH value of fermention medium is 7.0-7.2; The fermention medium of inoculation Geobacillus stearothermophilus temperature 55-65 DEG C, under mixing speed is 200-220r/min, enlarged culturing 24-40h, until the Geobacillus stearothermophilus spore production rate in fermentor tank is more than 80%, Geobacillus stearothermophilus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of Geobacillus stearothermophilus.
In addition, the invention allows for a kind of using method of high temperature degradation changing food waste microbiobacterial agent, its technical scheme is as follows:
The microbiobacterial agent prepared fully is mixed according to the ratio that mass ratio is 1:250-500 with changing food waste, at 55-65 DEG C, aerobic fermentation 12-15h.
Compared with prior art, the bacterial strain exerted than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus is carried out enlarged culturing by the present invention, carry out high-density enlarged culturing again, finally carry out adsorbing obtaining the single dry bacterium powder of each bacterial strain with carrier, rational proportion is adopted according to its purposes, obtain the high temperature microbe microbial inoculum for changing food waste of degrading after compound, for the process of changing food waste, it has following several respects advantage:
1, the changing food waste for heterogeneity can carry out rational proportion to single bacterium powder, each single bacterial strain is cooperated with each other, Synergistic degradation changing food waste.
2, in composite fungus agent, in each one-component, living bacteria count is high, for degrade changing food waste time, only need an a small amount of microbiobacterial agent just can to degrade fully, rapidly changing food waste, cost is low, consumption is few, using method is easy.
3, in composite fungus agent, each component is high-temperature strain, and optimum leavening temperature is all at 55-65 DEG C, and effectively can carry out deactivation to pathogenic bacterium such as the intestinal bacteria in changing food waste, streptococcus aureuses during the fermentation, biological safety is high.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail:
A kind of high temperature degradation changing food waste microbiobacterial agent, is composited by the strain raw material of following mass fraction: exert than halogen ground anaerobic genus bacillus 15-30 part, thermophilic liquefaction genus bacillus 10-25 part, hot denitrogenation ground bacillus 20-40 part, Geobacillus stearothermophilus 25-50 part.
In actual application, according to the difference of protein, starch, fat, content of cellulose in changing food waste, the consumption of above each component in microbiobacterial agent can be adjusted targetedly.
One, the preparation of the single bacterium powder of each bacterial strain and microbiobacterial agent
Embodiment 1
Individual plant cultivation and high-density enlarged culturing is carried out than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus respectively to exerting, finally the single thalline diatomite obtained after cultivation is adsorbed, the single dry bacterium powder of obtained each bacterial strain.
1. exert the preparation of bacterium powder more single than halogen ground anaerobic genus bacillus
(1), exerting on the test tube slant after purified is seeded to than halogen ground anaerobic genus bacillus volume ratio is housed is in the triangular flask of liquid activation medium of 10-20%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 8g/L, extractum carnis 3g/L, yeast extract paste 5g/L, Zulkovsky starch 17g/L, sodium-chlor 4g/L, and the pH value of liquid activation medium is 7.0; Inoculation exerts liquid activation medium than halogen ground anaerobic genus bacillus under temperature 55 DEG C, shaking speed are 140r/min, cultivates 22h;
(2), will exerting to be seeded to than halogen ground anaerobic genus bacillus and be equipped with in fermentor tank that volume ratio is the fermention medium of 65% through step (1) activation gained, inoculum size is 2%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8g/L, extractum carnis 3g/L, yeast extract paste 5g/L, Zulkovsky starch 17g/L, sodium-chlor 4g/L, and the pH value of liquid activation medium is 7.0; Inoculation exerts liquid activation medium than halogen ground anaerobic genus bacillus under temperature 55 DEG C, mixing speed are 200r/min, enlarged culturing 28h, until in fermentor tank to exert than halogen ground anaerobic genus bacillus spore production rate be more than 80%, exerting after fermentation is adsorbed, until living bacteria count reaches 5 × 10 than halogen ground anaerobic genus bacillus carrier
9individual/gram, the single bacterium powder than halogen ground anaerobic genus bacillus must be exerted.
2. the preparation of the single bacterium powder of thermophilic liquefaction genus bacillus
(1), the thermophilic liquefaction genus bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 15%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 10g/L, extractum carnis 4g/L, tween 80 25ml/L, N.F,USP MANNITOL 15g/L, yeast extract paste 9g/L, glucose 25g/L, sodium-chlor 6g/L, and the pH value of liquid activation medium is 7.1; The liquid activation medium inoculating thermophilic liquefaction genus bacillus, under temperature 60 C, shaking speed 160r/min, cultivates 18h;
(2), by being seeded to through the thermophilic liquefaction genus bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 70%, inoculum size is 3%; The solvent of fermention medium is distilled water, solute component and concentration thereof are: peptone 10g/L, extractum carnis 4g/L, tween 80 25ml/L, N.F,USP MANNITOL 15g/L, yeast extract paste 9g/L, glucose 25g/L, sodium-chlor 6g/L, and the pH value of liquid activation medium is 7.1; Inoculate the liquid activation medium of thermophilic liquefaction genus bacillus under temperature 60 C, mixing speed are 220r/min, enlarged culturing 24h, until the thermophilic liquefaction genus bacillus spore production rate in fermentor tank is more than 80%, thermophilic liquefaction genus bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of thermophilic liquefaction genus bacillus.
3. the preparation of the single bacterium powder of hot denitrogenation ground bacillus
(1), the denitrogenation ground bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 25%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 9g/L, extractum carnis 4g/L, N.F,USP MANNITOL 20g/L, glucose 25g/L, sodium-chlor 4g/L, and the pH value of liquid activation medium is 7.0; The liquid activation medium of inoculation denitrogenation ground bacillus, under temperature 65 DEG C, shaking speed are 150r/min, cultivates 19h;
(2), by being seeded to through the denitrogenation ground bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 70%, inoculum size is 2%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 11g/L, extractum carnis 3g/L, N.F,USP MANNITOL 22g/L, glucose 24g/L, sodium-chlor 5g/L, and the pH value of fermention medium is 7.0; The fermention medium of inoculation denitrogenation ground bacillus is under temperature 58 DEG C, mixing speed are 210r/min, enlarged culturing 36h, until the denitrogenation ground bacillus spore production rate in fermentor tank is more than 80%, denitrogenation ground bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of denitrogenation ground bacillus.
4. the preparation of the single bacterium powder of Geobacillus stearothermophilus
(1), the Geobacillus stearothermophilus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 15%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 9g/L, yeast extract paste 7g/L, glucose 18g/L, sodium-chlor 5g/L, Bromothymol blue solution trace, and the pH value of liquid activation medium is 7.2; The liquid activation medium of inoculation Geobacillus stearothermophilus, under temperature 62 DEG C, shaking speed are 150r/min, cultivates 24h;
(2), by being seeded to through the Geobacillus stearothermophilus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 75%, inoculum size is 4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 10g/L, yeast extract paste 9g/L, glucose 17g/L, sodium-chlor 5g/L, Bromothymol blue solution trace, and the pH value of fermention medium is 7.0; The fermention medium of inoculation Geobacillus stearothermophilus is under temperature 57 DEG C, mixing speed are 220r/min, enlarged culturing 40h, until the Geobacillus stearothermophilus spore production rate in fermentor tank is more than 80%, Geobacillus stearothermophilus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of Geobacillus stearothermophilus.
5. the preparation of microbiobacterial agent
Get and exert than halogen ground anaerobic genus bacillus 25 parts, thermophilic liquefaction genus bacillus 20 parts, hot denitrogenation ground bacillus 30 parts, Geobacillus stearothermophilus 40 parts, the mass fraction of single for above-mentioned each bacterial strain bacterium powder is fully mixed to get microbiobacterial agent.
Embodiment 2
Individual plant cultivation and high-density enlarged culturing is carried out than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus respectively to exerting, finally the single thalline diatomite obtained after cultivation is adsorbed, the single dry bacterium powder of obtained each bacterial strain.
1. exert the preparation of bacterium powder more single than halogen ground anaerobic genus bacillus
(1), exerting on the test tube slant after purified is seeded to than halogen ground anaerobic genus bacillus volume ratio is housed is in the triangular flask of liquid activation medium of 15%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 9g/L, extractum carnis 4g/L, yeast extract paste 3g/L, Zulkovsky starch 25g/L, sodium-chlor 6g/L, and the pH value of liquid activation medium is 7.0; Inoculation exerts liquid activation medium than halogen ground anaerobic genus bacillus under temperature 60 C, shaking speed are 160r/min, cultivates 18h;
(2), will exerting to be seeded to than halogen ground anaerobic genus bacillus and be equipped with in fermentor tank that volume ratio is the fermention medium of 60% through step (1) activation gained, inoculum size is 4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 10g/L, extractum carnis 3g/L, yeast extract paste 5g/L, Zulkovsky starch 18g/L, sodium-chlor 5g/L, and the pH value of fermention medium is 7.2; Inoculation exerts fermention medium than halogen ground anaerobic genus bacillus under temperature 65 DEG C, mixing speed are 210r/min, enlarged culturing 25h, until in fermentor tank to exert than halogen ground anaerobic genus bacillus spore production rate be more than 80%, exerting after fermentation is adsorbed, until living bacteria count reaches 5 × 10 than halogen ground anaerobic genus bacillus carrier
9individual/gram, the single bacterium powder than halogen ground anaerobic genus bacillus must be exerted.
2. the preparation of the single bacterium powder of thermophilic liquefaction genus bacillus
(1), the thermophilic liquefaction genus bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 20%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 8g/L, extractum carnis 4g/L, tween 80 20ml/L, N.F,USP MANNITOL 20g/L, yeast extract paste 7g/L, glucose 24g/L, sodium-chlor 4g/L, and the pH value of liquid activation medium is 7.1; The liquid activation medium inoculating thermophilic liquefaction genus bacillus, under temperature 55 DEG C, shaking speed 140r/min, cultivates 20h;
(2), by being seeded to through the thermophilic liquefaction genus bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 60%, inoculum size is 2%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8g/L, extractum carnis 4g/L, tween 80 20ml/L, N.F,USP MANNITOL 20g/L, yeast extract paste 7g/L, glucose 24g/L, sodium-chlor 4g/L, and the pH value of liquid activation medium is 7.1; Inoculate the liquid activation medium of thermophilic liquefaction genus bacillus under temperature 55 DEG C, mixing speed are 210r/min, enlarged culturing 26h, until the thermophilic liquefaction genus bacillus spore production rate in fermentor tank is more than 80%, thermophilic liquefaction genus bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of thermophilic liquefaction genus bacillus.
3. the preparation of the single bacterium powder of hot denitrogenation ground bacillus
(1), the denitrogenation ground bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 25%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 10g/L, extractum carnis 3g/L, N.F,USP MANNITOL 23g/L, glucose 24g/L, sodium-chlor 4g/L, and the pH value of liquid activation medium is 7.2; The liquid activation medium of inoculation denitrogenation ground bacillus, under temperature 65 DEG C, shaking speed are 160r/min, cultivates 22h;
(2), by being seeded to through the denitrogenation ground bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 62%, inoculum size is 3%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 10g/L, extractum carnis 3g/L, N.F,USP MANNITOL 23g/L, glucose 24g/L, sodium-chlor 4g/L, and the pH value of liquid activation medium is 7.2; The liquid activation medium of inoculation denitrogenation ground bacillus is under temperature 65 DEG C, mixing speed are 200r/min, enlarged culturing 28h, until the denitrogenation ground bacillus spore production rate in fermentor tank is more than 80%, denitrogenation ground bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of denitrogenation ground bacillus.
4. the preparation of the single bacterium powder of Geobacillus stearothermophilus
(1), the Geobacillus stearothermophilus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 20%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 12g/L, yeast extract paste 10g/L, glucose 15g/L, sodium-chlor 6g/L, Bromothymol blue solution trace, and the pH value of liquid activation medium is 7.2; The liquid activation medium of inoculation Geobacillus stearothermophilus, under temperature 55 DEG C, shaking speed are 160r/min, cultivates 24h;
(2), by being seeded to through the Geobacillus stearothermophilus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 75%, inoculum size is 4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 12g/L, yeast extract paste 10g/L, glucose 15g/L, sodium-chlor 6g/L, Bromothymol blue solution trace, and the pH value of liquid activation medium is 7.2; The liquid activation medium of inoculation Geobacillus stearothermophilus is under temperature 55 DEG C, mixing speed are 210r/min, enlarged culturing 40h, until the Geobacillus stearothermophilus spore production rate in fermentor tank is more than 80%, Geobacillus stearothermophilus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of Geobacillus stearothermophilus.
5. the preparation of microbiobacterial agent
Get and exert than halogen ground anaerobic genus bacillus 20 parts, thermophilic liquefaction genus bacillus 15 parts, hot denitrogenation ground bacillus 25 parts, Geobacillus stearothermophilus 30 parts, the mass fraction of single for above-mentioned each bacterial strain bacterium powder is fully mixed to get microbiobacterial agent.
Embodiment 3
Individual plant cultivation and high-density enlarged culturing is carried out than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus respectively to exerting, finally the single thalline diatomite obtained after cultivation is adsorbed, the single dry bacterium powder of obtained each bacterial strain.
1. exert the preparation of bacterium powder more single than halogen ground anaerobic genus bacillus
(1), exerting on the test tube slant after purified is seeded to than halogen ground anaerobic genus bacillus volume ratio is housed is in the triangular flask of liquid activation medium of 10%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 12g/L, extractum carnis 2g/L, yeast extract paste 4g/L, Zulkovsky starch 15g/L, sodium-chlor 5g/L, and the pH value of liquid activation medium is 7.1; Inoculation exerts liquid activation medium than halogen ground anaerobic genus bacillus under temperature 65 DEG C, shaking speed are 150r/min, cultivates 24h;
(2), will exerting to be seeded to than halogen ground anaerobic genus bacillus and be equipped with in fermentor tank that volume ratio is the fermention medium of 75% through step (1) activation gained, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 12g/L, extractum carnis 4g/L, yeast extract paste 3g/L, Zulkovsky starch 35g/L, sodium-chlor 6g/L, and the pH value of fermention medium is 7.1; Inoculation exert than halogen ground anaerobic genus bacillus fermention medium temperature 55-65 DEG C, under mixing speed is 220r/min, enlarged culturing 30h, until in fermentor tank to exert than halogen ground anaerobic genus bacillus spore production rate be more than 80%, exerting after fermentation is adsorbed, until living bacteria count reaches 5 × 10 than halogen ground anaerobic genus bacillus carrier
9individual/gram, the single bacterium powder than halogen ground anaerobic genus bacillus must be exerted.
2. the preparation of the single bacterium powder of thermophilic liquefaction genus bacillus
(1), the thermophilic liquefaction genus bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 12g/L, extractum carnis 2g/L, tween 80 35ml/L, N.F,USP MANNITOL 25g/L, yeast extract paste 10g/L, glucose 15g/L, sodium-chlor 5g/L, and the pH value of liquid activation medium is 7.2; The liquid activation medium inoculating thermophilic liquefaction genus bacillus, under temperature 60 C, shaking speed 155r/min, cultivates 24h;
(2), by being seeded to through the thermophilic liquefaction genus bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 75%, inoculum size is 4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 12g/L, extractum carnis 2g/L, tween 80 35ml/L, N.F,USP MANNITOL 25g/L, yeast extract paste 10g/L, glucose 15g/L, sodium-chlor 5g/L, and the pH value of fermention medium is 7.2; Inoculate the fermention medium of thermophilic liquefaction genus bacillus under temperature 62 DEG C, mixing speed are 200r/min, enlarged culturing 36h, until the thermophilic liquefaction genus bacillus spore production rate in fermentor tank is more than 80%, thermophilic liquefaction genus bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of thermophilic liquefaction genus bacillus.
3. the preparation of the single bacterium powder of hot denitrogenation ground bacillus
(1), the denitrogenation ground bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 12g/L, extractum carnis 2g/L, N.F,USP MANNITOL 25g/L, glucose 15g/L, sodium-chlor 6g/L, and the pH value of liquid activation medium is 7.1; The liquid activation medium of inoculation denitrogenation ground bacillus, under temperature 55 DEG C, shaking speed are 140r/min, cultivates 24h;
(2), by being seeded to through the denitrogenation ground bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 75%, inoculum size is 4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 12g/L, extractum carnis 4g/L, N.F,USP MANNITOL 25g/L, glucose 25g/L, sodium-chlor 6g/L, and the pH value of fermention medium is 7.1; The fermention medium of inoculation denitrogenation ground bacillus is under temperature 55 DEG C, mixing speed are 220r/min, enlarged culturing 40h, until the denitrogenation ground bacillus spore production rate in fermentor tank is more than 80%, denitrogenation ground bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of denitrogenation ground bacillus.
4. the preparation of the single bacterium powder of Geobacillus stearothermophilus
(1), the Geobacillus stearothermophilus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 8g/L, yeast extract paste 5g/L, glucose 25g/L, sodium-chlor 4g/L, Bromothymol blue solution trace, and the pH value of liquid activation medium is 7.0; The liquid activation medium of inoculation Geobacillus stearothermophilus, under temperature 65 DEG C, shaking speed are 140r/min, cultivates 18h;
(2), by being seeded to through the Geobacillus stearothermophilus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 60%, inoculum size is 2%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8g/L, yeast extract paste 5g/L, glucose 25g/L, sodium-chlor 4g/L, Bromothymol blue solution trace, and the pH value of fermention medium is 7.1; The fermention medium of inoculation Geobacillus stearothermophilus is under temperature 65 DEG C, mixing speed are 200r/min, enlarged culturing 24h, until the Geobacillus stearothermophilus spore production rate in fermentor tank is more than 80%, Geobacillus stearothermophilus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of Geobacillus stearothermophilus.
(2), by being seeded to through the Geobacillus stearothermophilus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 68%, inoculum size is 3%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 9g/L, yeast extract paste 8g/L, glucose 18g/L, sodium-chlor 5g/L, Bromothymol blue solution trace, and the pH value of fermention medium is 7.0; The fermention medium of inoculation Geobacillus stearothermophilus is under temperature 58 DEG C, mixing speed are 210r/min, enlarged culturing 36h, until the Geobacillus stearothermophilus spore production rate in fermentor tank is more than 80%, Geobacillus stearothermophilus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of Geobacillus stearothermophilus.
5. the preparation of microbiobacterial agent
Get and exert than halogen ground anaerobic genus bacillus 30 parts, thermophilic liquefaction genus bacillus 25 parts, hot denitrogenation ground bacillus 40 parts, Geobacillus stearothermophilus 50 parts, the mass fraction of single for above-mentioned each bacterial strain bacterium powder is fully mixed to get microbiobacterial agent.
Embodiment 4
Individual plant cultivation and high-density enlarged culturing is carried out than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus respectively to exerting, finally the single thalline diatomite obtained after cultivation is adsorbed, the single dry bacterium powder of obtained each bacterial strain.
1. exert the preparation of bacterium powder more single than halogen ground anaerobic genus bacillus
(1), exerting on the test tube slant after purified is seeded to than halogen ground anaerobic genus bacillus volume ratio is housed is in the triangular flask of liquid activation medium of 20%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 10g/L, extractum carnis 4g/L, yeast extract paste 5g/L, Zulkovsky starch 35g/L, sodium-chlor 6g/L, and the pH value of liquid activation medium is 7.0; Inoculation exerts liquid activation medium than halogen ground anaerobic genus bacillus under temperature 60 C, shaking speed are 155r/min, cultivates 20h;
(2), will exerting to be seeded to than halogen ground anaerobic genus bacillus and be equipped with in fermentor tank that volume ratio is the fermention medium of 70% through step (1) activation gained, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 11g/L, extractum carnis 2g/L, yeast extract paste 4g/L, Zulkovsky starch 15g/L, sodium-chlor 5g/L, and the pH value of fermention medium is 7.0-7.2; Inoculation exert than halogen ground anaerobic genus bacillus fermention medium temperature 55-65 DEG C, under mixing speed is 205r/min, enlarged culturing 20h, until in fermentor tank to exert than halogen ground anaerobic genus bacillus spore production rate be more than 80%, exerting after fermentation is adsorbed, until living bacteria count reaches 5 × 10 than halogen ground anaerobic genus bacillus carrier
9individual/gram, the single bacterium powder than halogen ground anaerobic genus bacillus must be exerted.
2. the preparation of the single bacterium powder of thermophilic liquefaction genus bacillus
(1), the thermophilic liquefaction genus bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 18%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 9g/L, extractum carnis 3g/L, tween 80 28ml/L, N.F,USP MANNITOL 20g/L, yeast extract paste 5g/L, glucose 20g/L, sodium-chlor 5g/L, and the pH value of liquid activation medium is 7.0; The liquid activation medium inoculating thermophilic liquefaction genus bacillus, under temperature 57 DEG C, shaking speed 150r/min, cultivates 21h;
(2), by being seeded to through the thermophilic liquefaction genus bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 70%, inoculum size is 3%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 11g/L, extractum carnis 3g/L, tween 80 30ml/L, N.F,USP MANNITOL 20g/L, yeast extract paste 5g/L, glucose 15g/L, sodium-chlor 6g/L, and the pH value of fermention medium is 7.0; Inoculate the fermention medium of thermophilic liquefaction genus bacillus under temperature 58 DEG C, mixing speed are 210r/min, enlarged culturing 30h, until the thermophilic liquefaction genus bacillus spore production rate in fermentor tank is more than 80%, thermophilic liquefaction genus bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of thermophilic liquefaction genus bacillus.
3. the preparation of the single bacterium powder of hot denitrogenation ground bacillus
(1), the denitrogenation ground bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 18%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 8g/L, extractum carnis 3g/L, N.F,USP MANNITOL 15g/L, glucose 23g/L, sodium-chlor 5g/L, and the pH value of liquid activation medium is 7.0; The liquid activation medium of inoculation denitrogenation ground bacillus, under temperature 60 C, shaking speed are 150r/min, cultivates 18h;
(2), by being seeded to through the denitrogenation ground bacillus of step (1) activation gained be equipped with in fermentor tank that volume ratio is the fermention medium of 60%, inoculum size is 3%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8g/L, extractum carnis 2g/L, N.F,USP MANNITOL 15g/L, glucose 15g/L, sodium-chlor 5g/L, and the pH value of fermention medium is 7.0; The fermention medium of inoculation denitrogenation ground bacillus is under temperature 58 DEG C, mixing speed are 210r/min, enlarged culturing 24h, until the denitrogenation ground bacillus spore production rate in fermentor tank is more than 80%, denitrogenation ground bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of denitrogenation ground bacillus.
4. the preparation of the single bacterium powder of Geobacillus stearothermophilus
(1), the Geobacillus stearothermophilus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 18%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 9g/L, yeast extract paste 8g/L, glucose 23g/L, sodium-chlor 5g/L, Bromothymol blue solution trace, and the pH value of liquid activation medium is 7.1; The liquid activation medium of inoculation Geobacillus stearothermophilus, under temperature 58 DEG C, shaking speed are 150r/min, cultivates 21h;
5. the preparation of microbiobacterial agent
Get and exert than halogen ground anaerobic genus bacillus 15 parts, thermophilic liquefaction genus bacillus 10 parts, hot denitrogenation ground bacillus 20 parts, Geobacillus stearothermophilus 25 parts, the mass fraction of single for above-mentioned each bacterial strain bacterium powder is fully mixed to get microbiobacterial agent.
Take four kinds of microbiobacterial agents that embodiment 1-4 is obtained, be seeded to respectively in the changing food waste of actual acquisition, observe the effect that microbial inoculum plays in actual applications, measure each index reduction rate of selected changing food waste.
Two, experimental technique
For examination changing food waste: initial water content is 49.24%, Contents of Main Components (in butt) is starch 49.14%, protein 17.51%, fat 22.27%, Mierocrystalline cellulose 1.47%, other 9.61%; Above-mentioned content is mass percent.
Get for examination changing food waste 5 parts, 300 kilograms every part, get the microbiobacterial agent 900g/ kind prepared in example 1-4; A microbial inoculum is corresponding aly puts into the integrated treatment facility of changing food waste together for examination changing food waste, after mixing at 55 DEG C aerobic fermentation 12h; The reduction rate for each component in examination changing food waste is measured, experiment repetition 3 times after fermentation ends.
Three, experimental result
1. adopt the experimental result of embodiment 1 microbiobacterial agent
(1), after measured, examination changing food waste reduction rate is in mass supplied to be 91.75% before and after process;
(2), after measured, for trying, starch reduction rate in naval vessels boats and ships changing food waste is 86.42%, protein reduction rate is 91.23%, fatty reduction rate is 84.96%, Mierocrystalline cellulose reduction rate is 89.57%.
2. adopt the experimental result of embodiment 2 microbiobacterial agent
(1), after measured, examination changing food waste reduction rate is in mass supplied to be 89.37% before and after process;
(2), after measured, for trying, starch reduction rate in changing food waste is 79.20%, protein reduction rate is 89.54%, fatty reduction rate is 80.73%, Mierocrystalline cellulose reduction rate is 97.62%.
3. adopt the experimental result of embodiment 3 microbiobacterial agent
(1), after measured, examination changing food waste reduction rate is in mass supplied to be 90.15% before and after process;
(2), after measured, for trying, starch reduction rate in changing food waste is 83.24%, protein reduction rate is 92.15%, fatty reduction rate is 89.12%, Mierocrystalline cellulose reduction rate is 88.75%.
4. adopt the experimental result of embodiment 4 microbiobacterial agent
(1), after measured, examination changing food waste reduction rate is in mass supplied to be 89.65% before and after process;
(2), after measured, for trying, starch reduction rate in changing food waste is 91.20%, protein reduction rate is 86.79%, fatty reduction rate is 85.46%, Mierocrystalline cellulose reduction rate is 91.02%.
Above for testing the more excellent proportioning of the changing food waste setting provided for this, according to the difference of protein, starch, fat, content of cellulose in changing food waste, the consumption of each component in microbial inoculum can be adjusted targetedly in the application process of reality.
In addition, when utilizing high temperature microbe facture to carry out fermentative processing to changing food waste, the mass ratio of microbiobacterial agent and changing food waste is not limited to above-mentioned citing, as long as in the quality of 1:250-500 than in scope, thermophilic fermentation temperature and aerobic fermentation time are also not limited to above-mentioned citing, as long as control within the scope of 55-65 DEG C, 12-15h.Those skilled in the art can determine mass ratio, the temperature of reaction and fermentation time etc. of microbiobacterial agent and changing food waste flexibly according to actual changing food waste situation.
Certainly; more than illustrate and be only preferred embodiment of the present invention; the present invention is not limited to enumerate above-described embodiment; should be noted that; any those of ordinary skill in the art are under the instruction of this specification sheets; made all equivalently to substitute, obvious variant, within the essential scope all dropping on this specification sheets, protection of the present invention ought to be subject to.
Claims (7)
1. a high temperature degradation changing food waste microbiobacterial agent, it is characterized in that, described microbial strains is composited by the strain raw material of following mass fraction: exert than halogen ground anaerobic genus bacillus 15-30 part, thermophilic liquefaction genus bacillus 10-25 part, hot denitrogenation ground bacillus 20-40 part, Geobacillus stearothermophilus 25-50 part.
2. a preparation method for high temperature degradation changing food waste microbiobacterial agent, is characterized in that, comprises following preparation process:
S1, will exert and carry out individual plant cultivation respectively than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus, then obtained individual plant bacterium colony is carried out activation culture according in the inoculum size access liquid nutrient medium of 10-20%, then volume ratio is housed is ferment in the fermentation equipment of 60-75% substratum by the bacterial classification access after activation;
Adsorbed by gained fermented liquid carrier, obtained living bacteria count is 5 × 10
9individual/gram exert bacterium powder more single than halogen ground anaerobic genus bacillus, thermophilic liquefaction genus bacillus, hot denitrogenation ground bacillus, Geobacillus stearothermophilus; Absorption carrier is diatomite;
S2, obtain and exert than halogen ground anaerobic genus bacillus 15-30 part, thermophilic liquefaction genus bacillus 10-25 part, hot denitrogenation ground bacillus 20-40 part, Geobacillus stearothermophilus 25-50 part, the mass fraction of single for above-mentioned each bacterial strain bacterium powder is fully mixed to get microbiobacterial agent.
3. the preparation method of a kind of high temperature degradation changing food waste microbiobacterial agent according to claim 2, is characterized in that, in described step s1, exerts and than the single bacterium powder preparation process of halogen ground anaerobic genus bacillus is:
S111, exerting on the test tube slant after purified be seeded to than halogen ground anaerobic genus bacillus volume ratio is housed is in the triangular flask of liquid activation medium of 10-20%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, yeast extract paste 3-5g/L, Zulkovsky starch 15-35g/L, sodium-chlor 4-6g/L, and the pH value of liquid activation medium is 7.0-7.2; Inoculation exert than halogen ground anaerobic genus bacillus liquid activation medium temperature 55-65 DEG C, under shaking speed is 140-160r/min, cultivate 18-24h;
S112, will activate exerting to be seeded to than halogen ground anaerobic genus bacillus and being equipped with in fermentor tank that volume ratio is the fermention medium of 60-75% of gained through step s111, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, yeast extract paste 3-5g/L, Zulkovsky starch 15-35g/L, sodium-chlor 4-6g/L, and the pH value of fermention medium is 7.0-7.2; Inoculation exert than halogen ground anaerobic genus bacillus fermention medium temperature 55-65 DEG C, under mixing speed is 200-220r/min, enlarged culturing 20-30h, until in fermentor tank to exert than halogen ground anaerobic genus bacillus spore production rate be more than 80%, exerting after fermentation is adsorbed, until living bacteria count reaches 5 × 10 than halogen ground anaerobic genus bacillus carrier
9individual/gram, the single bacterium powder than halogen ground anaerobic genus bacillus must be exerted.
4. the preparation method of a kind of high temperature degradation changing food waste microbiobacterial agent according to claim 2, is characterized in that, in described step s1, the single bacterium powder preparation process of thermophilic liquefaction genus bacillus is:
S121, the thermophilic liquefaction genus bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10-20%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, tween 80 20-35ml/L, N.F,USP MANNITOL 15-25g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of liquid activation medium is 7.0-7.2; The liquid activation medium inoculating thermophilic liquefaction genus bacillus temperature 55-60 DEG C, under shaking speed 140-160r/min, cultivate 18-24h;
S122, to be seeded to by the thermophilic liquefaction genus bacillus activating gained to be equipped with in fermentor tank that volume ratio is the fermention medium of 60-75% through step s121, inoculum size is 2-4%; The solvent of fermention medium is distilled water, solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, tween 80 20-35ml/L, N.F,USP MANNITOL 15-25g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of fermention medium is 7.0-7.2; The fermention medium inoculating thermophilic liquefaction genus bacillus temperature 55-62 DEG C, under mixing speed is 200-220r/min, enlarged culturing 24-36h, until the thermophilic liquefaction genus bacillus spore production rate in fermentor tank is more than 80%, thermophilic liquefaction genus bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of thermophilic liquefaction genus bacillus.
5. the preparation method of a kind of high temperature degradation changing food waste microbiobacterial agent according to claim 2, is characterized in that, in described step s1, the single bacterium powder preparation process of hot denitrogenation ground bacillus is:
S131, the denitrogenation ground bacillus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10-25%; The solvent of liquid activation medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, N.F,USP MANNITOL 15-25g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of liquid activation medium is 7.0-7.2; The liquid activation medium of inoculation denitrogenation ground bacillus temperature 55-65 DEG C, under shaking speed is 140-160r/min, cultivate 18-24h;
S132, be seeded to be equipped with activating the denitrogenation ground bacillus of gained through step s131 in fermentor tank that volume ratio is the fermention medium of 60-75%, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, extractum carnis 2-4g/L, N.F,USP MANNITOL 15-25g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, and the pH value of fermention medium is 7.0-7.2; The fermention medium of inoculation denitrogenation ground bacillus temperature 55-65 DEG C, under mixing speed is 200-220r/min, enlarged culturing 24-40h, until the denitrogenation ground bacillus spore production rate in fermentor tank is more than 80%, denitrogenation ground bacillus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of denitrogenation ground bacillus.
6. the preparation method of a kind of high temperature degradation changing food waste microbiobacterial agent according to claim 2, is characterized in that, in described step s1, the single bacterium powder preparation process of Geobacillus stearothermophilus is:
S141, the Geobacillus stearothermophilus on the test tube slant after purified is seeded to volume ratio is housed is in the triangular flask of liquid activation medium of 10-20%; The solvent of liquid activation medium is distilled water, solute component and concentration thereof are: peptone 8-12g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, Bromothymol blue solution trace, and the pH value of liquid activation medium is 7.0-7.2; The liquid activation medium of inoculation Geobacillus stearothermophilus temperature 55-65 DEG C, under shaking speed is 140-160r/min, cultivate 18-24h;
S142, be seeded to be equipped with activating the Geobacillus stearothermophilus of gained through step s141 in fermentor tank that volume ratio is the fermention medium of 60-75%, inoculum size is 2-4%; The solvent of fermention medium is distilled water, and solute component and concentration thereof are: peptone 8-12g/L, yeast extract paste 5-10g/L, glucose 15-25g/L, sodium-chlor 4-6g/L, Bromothymol blue solution trace, and the pH value of fermention medium is 7.0-7.2; The fermention medium of inoculation Geobacillus stearothermophilus temperature 55-65 DEG C, under mixing speed is 200-220r/min, enlarged culturing 24-40h, until the Geobacillus stearothermophilus spore production rate in fermentor tank is more than 80%, Geobacillus stearothermophilus carrier after fermentation is adsorbed, until living bacteria count reaches 5 × 10
9individual/gram, obtain the single bacterium powder of Geobacillus stearothermophilus.
7. the using method of a high temperature degradation changing food waste microbiobacterial agent, adopt high temperature degradation changing food waste microbiobacterial agent as claimed in claim 1, it is characterized in that, microbiobacterial agent is fully mixed according to the ratio that mass ratio is 1:250-500 with changing food waste, at 55-65 DEG C, aerobic fermentation 12-15h.
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