CN103272361A - Active biological complexing agent for degradation and digestion of organic wet garbage and preparation method of active biological complexing agent - Google Patents
Active biological complexing agent for degradation and digestion of organic wet garbage and preparation method of active biological complexing agent Download PDFInfo
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Abstract
The invention relates to an active biological complexing agent for degradation and digestion of organic wet garbage and a preparation method of the active biological complexing agent. The active biological complexing agent comprises the following components in percentage by weight: 0.6-1.0 percent of pichia jadinii solution, 0.6-1.0 percent of Japanese aspergillus solution, 0.6-1.0 percent of aspergillus terreus solution, 0.6-1.0 percent of bacillus cereus solution, 0.6-4.0 percent of Bacillus thermoliquefaciens-NL solution, 0.6-1.0 percent of geobacillus stearothermophilus solution, 0.6-1.0 percent of aspergillus nidulans solution, 50-60 percent of rice husks, 4-5 percent of swill and the balance of water. The active biological complexing agent for degradation and digestion of the organic wet garbage is prepared by carrying out mixed fermentation and domestication. Compared with the prior art, the active biological complexing agent organically combines the solutions together, has the capability of decomposing wet garbage into gases, water and inorganic salts within 8-24 hours and has the degradation rate of 75-99.9 percent; and no external heating is needed.
Description
Technical field
The present invention relates to the activating agent of organic waste and preparation method thereof of degrading, especially relate to a kind of organic wet refuse degraded active bio-compounding agent and preparation method thereof of dissolving.
Background technology
The known biology that rises is in the world strengthened (input) technology, and its principle is to utilize the method for biological culture, selects single culture, is prepared into the higher liquid preparation of bacteria containing amount, renders in the engineering interface, in order to the strength of enhanced biological degraded.But its preparation cost height, production capacity are low, liquid bacterial agent in engineering interface term of validity weak point, poor stability, that condition of storage harshness, engineering are used the fortune consumption in addition is excessive.Lack industrialization prospect, the most fatal defective is that the bacterial classification natural quality of liquid bacterial agent is at all constant, can't solve fundamentally that resistance to soiling is weak, environmental resistance difference and the low problem of decomposition rate.
Aspect the degraded of the surplus organic waste in kitchen dissolved, domestic technique was single, bacterial classification is single, and external microbial inoculum can't adapt to Chinese food and drink fiber crops, characteristics such as peppery again, and the prior biological technology all needs external heat to assist, power consumption is big, and it is not thorough to degrade, and stink is strong during operation.
Summary of the invention
Purpose of the present invention is exactly to provide a kind of in order to overcome the defective that above-mentioned prior art exists organic wet refuse is resolved into gas, water and inorganic salts, and degradation rate reaches organic wet refuse degraded of 75～99.9% active bio-compounding agent and preparation method thereof of dissolving.
Purpose of the present invention can be achieved through the following technical solutions:
A kind of organic wet refuse degraded active bio-compounding agent of dissolving, comprise following component and weight percent content: outstanding fourth Pichia yeast liquid 0.6～1.0%, koji mould liquid 0.6～1.0%, aspergillus terreus liquid 0.6～1.0%, bacillus cereus bacterium liquid 0.6～1.0%, thermophilic liquefaction bacillus bacterium liquid 0.6～1.0%, stearothermophilus ground bacillus bacterium liquid 0.6～1.0%, aspergillus nidulans liquid 0.6～1.0%, husk 50～60%, swill 4～5%, surplus is water.
The bacteria containing amount of described outstanding fourth Pichia yeast liquid is 1 * 10
8～9 * 10
12Individual/ml.
The bacteria containing amount of described koji mould liquid is 1 * 10
8～9 * 10
12Individual/ml
The bacteria containing amount of described aspergillus terreus liquid is 1 * 10
8～9 * 10
12Individual/ml.
The bacteria containing amount of described bacillus cereus bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
The bacteria containing amount of described thermophilic liquefaction bacillus bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
The bacteria containing amount of described stearothermophilus ground bacillus bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
The bacteria containing amount of described aspergillus nidulans liquid is 1 * 10
8～9 * 10
12Individual/ml.
The dissolve preparation method of active bio-compounding agent may further comprise the steps the degraded of organic wet refuse: the preparation of (1) outstanding fourth Pichia yeast liquid:
A, the outstanding fourth Pichia pastoris culture medium raw material of preparation comprise following component and weight percent content: agar 1.4～1.6%, 5 ° of brewer's worts 98.4～98.6%;
The preparation of b, fluid nutrient medium: by outstanding fourth Pichia pastoris culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized outstanding fourth Pichia pastoris culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane outstanding fourth Pichia pastoris bacterial classification and insert in gnotobasis in the triangular flask that contains the outstanding fourth Pichia pastoris culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of outstanding fourth Pichia pastoris culture medium, at 30～45 ℃, ventilating ratio is 1: 1～2 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the outstanding fourth Pichia pastoris culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain outstanding fourth Pichia yeast liquid;
(2) preparation of koji mould liquid:
A, preparation aspergillus japonicus culture medium raw material, comprise following component and weight percent content: sucrose 2.8～3.0%, NaNO30.28～0.3%, MgSO47H2O0.045～0.048%, KCl0.0045～0.0048%, FeSO44H2O0.001%, K2HPO40.095～0.096%, agar 1.4%～1.44%, surplus is distilled water, and pH is 6.0～6.5;
The preparation of b, fluid nutrient medium: by Japanese aspergillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus japonicus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus japonicus bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus japonicus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus japonicus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus japonicus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain koji mould liquid;
(3) preparation of aspergillus terreus liquid:
A, preparation Aspergillus terreus culture medium raw material, comprise following component and weight percent content: sucrose 2.8～3.0%, NaNO30.28～0.3%, MgSO47H2O0.045～0.048%, KCl0.0045～0.0048%, FeSO44H2O0.001%, K2HPO40.095～0.096%, agar 1.4%～1.44%, surplus is distilled water, and pH is 6.0～6.5;
The preparation of b, fluid nutrient medium: press Aspergillus terreus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized Aspergillus terreus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane Aspergillus terreus bacterial classification and insert in gnotobasis in the triangular flask that contains the Aspergillus terreus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of Aspergillus terreus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the Aspergillus terreus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain aspergillus terreus liquid;
(4) preparation of bacillus cereus bacterium liquid:
A, preparation bacillus cereus culture medium raw material comprise following component and weight percent content: peptone 0.4～0.6%, beef leaching thing 0.25～0.35%, NaCl0.4～0.6%, agar 1.4～1.5%, and surplus is distilled water;
The preparation of b, fluid nutrient medium: by bacillus cereus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized bacillus cereus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane bacillus cereus bacterial classification and insert in gnotobasis in the triangular flask that contains the bacillus cereus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of bacillus cereus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the bacillus cereus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain bacillus cereus bacterium liquid;
(5) preparation of thermophilic liquefaction bacillus bacterium liquid:
A, the thermophilic liquefaction bacillus culture medium raw material of preparation, comprise following component and weight percent content: peptone 0.45～0.5%, beef leaching thing 0.27～0.30%, NaCl0.45～0.5%, agar 1.45～1.50%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: by thermophilic liquefaction bacillus culture medium formulated raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized thermophilic liquefaction bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get the thermophilic liquefaction in inclined-plane bacillus bacterial classification and in gnotobasis, insert in the triangular flask that contains the thermophilic liquefaction bacillus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of thermophilic liquefaction bacillus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the thermophilic liquefaction bacillus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain thermophilic liquefaction bacillus bacterium liquid;
(6) preparation of stearothermophilus ground bacillus bacterium liquid:
A, preparation stearothermophilus ground bacillus culture medium raw material, comprise following component and weight percent content: peptone 0.45～0.5%, beef leaching thing 0.27～0.30%, NaCl0.45～0.5%, agar 1.45～1.50%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: press stearothermophilus ground bacillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized stearothermophilus ground bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane stearothermophilus ground bacillus bacterial classification and in gnotobasis, insert in the triangular flask that contains the stearothermophilus ground bacillus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of stearothermophilus ground bacillus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the stearothermophilus ground bacillus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain stearothermophilus ground bacillus bacterium liquid;
(7) preparation of aspergillus nidulans liquid:
A, preparation aspergillus nidulans culture medium raw material, comprise following component and weight percent content: agar 1.45～1.48%, surplus are the 12Brix. brewer's wort;
The preparation of b, fluid nutrient medium: press aspergillus nidulans culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus nidulans culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus nidulans bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus nidulans culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus nidulans culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the aspergillus nidulans culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain aspergillus nidulans liquid;
(8) with outstanding fourth Pichia yeast liquid, koji mould liquid, aspergillus terreus liquid, bacillus cereus bacterium liquid, thermophilic liquefaction bacillus bacterium liquid, stearothermophilus ground bacillus bacterium liquid, aspergillus nidulans liquid is puddled in husk by prescription, adding and keeping humidity behind the water is 40～60%, windrow covers fermentation, when piling temperature between 40～60 ℃, husk is moved to fermentation tank continue fermentation, and the 30～60min/3h that ventilates, every 12h turning once, control heap temperature is at 50～70 ℃ simultaneously, humidity is 20～50%, shakeout the stockpile cooling behind fermentation 7～10d, below the heap temperature drop to 20 ℃, humidity is down to 10～20%, utilize swill to build the working environment of the organic wet refuse degraded of simulation for mixed bacterium liquid at last, tame seven kinds of bacterial classifications to the adaptability of organic wet refuse and each other with the suitable natural disposition of indigenous bacterial classification, namely prepare organic wet refuse degraded active bio-compounding agent of dissolving.
Compared with prior art, the fiber-like discarded object in organic wet refuse can be effectively decomposed in the combination of the outstanding fourth Pichia pastoris of the present invention's employing, aspergillus japonicus, thermophilic liquefaction bacillus; Carbohydrate in organic wet refuse, protein-based discarded object can be effectively decomposed in the combination of Aspergillus terreus, bacillus cereus, and the stearothermophilus ground bacillus can effectively decompose fats discarded object in organic wet refuse; Aspergillus nidulans can decompose the grease class discarded object in organic wet refuse, and all finally be decomposed into gas, water and grey matter, by above-mentioned bacterium liquid is organically combined, can in 8～24h, organic wet refuse be resolved into gas, water and inorganic salts, degradation rate reaches 75～99.9%, and need not external heat.
The specific embodiment
The present invention is described in detail below in conjunction with specific embodiment.
Embodiment 1
A kind of organic wet refuse degraded active bio-compounding agent of dissolving, comprise following component and weight percent content: outstanding fourth Pichia yeast liquid 0.6%, koji mould liquid 0.6%, aspergillus terreus liquid 0.6%, bacillus cereus bacterium liquid 0.6%, thermophilic liquefaction bacillus bacterium liquid 0.6%, stearothermophilus ground bacillus bacterium liquid 0.6%, aspergillus nidulans liquid 0.6%, husk 50%, swill 4%, surplus is water.Bacteria containing amount in the above-mentioned bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
The dissolve preparation method of active bio-compounding agent may further comprise the steps the degraded of organic wet refuse:
(1) preparation of outstanding fourth Pichia yeast liquid:
A, the outstanding fourth Pichia pastoris culture medium raw material of preparation comprise following component and weight percent content: agar 1.4%, 5 ° of brewer's worts 98.6%;
The preparation of b, fluid nutrient medium: by outstanding fourth Pichia pastoris culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized outstanding fourth Pichia pastoris culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane outstanding fourth Pichia pastoris bacterial classification and insert in gnotobasis in the triangular flask that contains the outstanding fourth Pichia pastoris culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt% contains the access of triangular flask strain liquid in the fermentation tank of outstanding fourth Pichia pastoris culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the outstanding fourth Pichia pastoris culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain outstanding fourth Pichia yeast liquid;
(2) preparation of koji mould liquid:
A, preparation aspergillus japonicus culture medium raw material, comprise following component and weight percent content: sucrose 2.8%, NaNO30.28%, MgSO47H2O0.045%, KCl0.0045%, FeSO44H2O0.001%, K2HPO40.095%, agar 1.4%%, surplus is distilled water, and pH is controlled to be 6.0;
The preparation of b, fluid nutrient medium: by Japanese aspergillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus japonicus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus japonicus bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus japonicus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus japonicus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus japonicus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain koji mould liquid;
(3) preparation of aspergillus terreus liquid:
A, preparation Aspergillus terreus culture medium raw material, comprise following component and weight percent content: sucrose 2.8%, NaNO30.28%, MgSO47H2O0.045%, KCl0.0045%, FeSO44H2O0.001%, K2HPO40.095%, agar 1.4%, surplus is distilled water, and pH is 6.0;
The preparation of b, fluid nutrient medium: press Aspergillus terreus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized Aspergillus terreus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane Aspergillus terreus bacterial classification and insert in gnotobasis in the triangular flask that contains the Aspergillus terreus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt% contains the access of triangular flask strain liquid in the fermentation tank of Aspergillus terreus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the Aspergillus terreus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain aspergillus terreus liquid;
(4) preparation of bacillus cereus bacterium liquid:
A, preparation bacillus cereus culture medium raw material comprise following component and weight percent content: peptone 0.4%, beef leaching thing 0.25%, NaCl0.4%, agar 1.4%, and surplus is distilled water;
The preparation of b, fluid nutrient medium: by bacillus cereus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized bacillus cereus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane bacillus cereus bacterial classification and insert in gnotobasis in the triangular flask that contains the bacillus cereus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt% contains the access of triangular flask strain liquid in the fermentation tank of bacillus cereus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the bacillus cereus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain bacillus cereus bacterium liquid;
(5) preparation of thermophilic liquefaction bacillus bacterium liquid:
A, the thermophilic liquefaction bacillus culture medium raw material of preparation comprise following component and weight percent content: peptone 0.45%, beef leaching thing 0.27%, NaCl0.45%, agar 1.45%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: by thermophilic liquefaction bacillus culture medium formulated raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized thermophilic liquefaction bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get the thermophilic liquefaction in inclined-plane bacillus bacterial classification and insert in gnotobasis in the triangular flask that contains the thermophilic liquefaction bacillus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt% contains the access of triangular flask strain liquid in the fermentation tank of thermophilic liquefaction bacillus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the thermophilic liquefaction bacillus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain thermophilic liquefaction bacillus bacterium liquid;
(6) preparation of stearothermophilus ground bacillus bacterium liquid:
A, preparation stearothermophilus ground bacillus culture medium raw material comprise following component and weight percent content: peptone 0.45%, beef leaching thing 0.27%, NaCl0.45%, agar 1.45%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: press stearothermophilus ground bacillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized stearothermophilus ground bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane stearothermophilus ground bacillus bacterial classification and insert in gnotobasis in the triangular flask that contains the stearothermophilus ground bacillus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt% contains the access of triangular flask strain liquid in the fermentation tank of stearothermophilus ground bacillus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the stearothermophilus ground bacillus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain stearothermophilus ground bacillus bacterium liquid;
(7) preparation of aspergillus nidulans liquid:
A, preparation aspergillus nidulans culture medium raw material, comprise following component and weight percent content: agar 1.45%, surplus are the 12Brix. brewer's wort;
The preparation of b, fluid nutrient medium: press aspergillus nidulans culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus nidulans culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus nidulans bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus nidulans culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus nidulans culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus nidulans culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain aspergillus nidulans liquid;
(8) with outstanding fourth Pichia yeast liquid, koji mould liquid, aspergillus terreus liquid, bacillus cereus bacterium liquid, thermophilic liquefaction bacillus bacterium liquid, stearothermophilus ground bacillus bacterium liquid, aspergillus nidulans liquid is puddled in husk by prescription, adding and keeping humidity behind the water is 40%, windrow covers fermentation, when piling temperature greater than 40 ℃, husk is moved to fermentation tank continue fermentation, and ventilation 30min/3h, every 12h turning once, control heap temperature is at 50 ℃ simultaneously, humidity is 20%, shakeout the stockpile cooling behind the fermentation 7d, below the heap temperature drop to 20 ℃, humidity is down to 15%, utilizes swill to build the working environment of the organic wet refuse degraded of simulation for mixed bacterium liquid at last, tame seven kinds of bacterial classifications to the adaptability of organic wet refuse and each other with the suitable natural disposition of indigenous bacterial classification, namely prepare organic wet refuse degraded active bio-compounding agent of dissolving.
Embodiment 2
A kind of organic wet refuse degraded active bio-compounding agent of dissolving, comprise following component and weight percent content: outstanding fourth Pichia yeast liquid 0.8%, koji mould liquid 0.8%, aspergillus terreus liquid 0.8%, bacillus cereus bacterium liquid 0.8%, thermophilic liquefaction bacillus bacterium liquid 0.8%, stearothermophilus ground bacillus bacterium liquid 0.8%, aspergillus nidulans liquid 0.8%, husk 60%, swill 4%, surplus is water.Bacteria containing amount in the above-mentioned bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
The dissolve preparation method of active bio-compounding agent may further comprise the steps the degraded of organic wet refuse:
(1) preparation of outstanding fourth Pichia yeast liquid:
A, the outstanding fourth Pichia pastoris culture medium raw material of preparation comprise following component and weight percent content: agar 1.5%, 5 ° of brewer's worts 98.5%;
The preparation of b, fluid nutrient medium: by outstanding fourth Pichia pastoris culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized outstanding fourth Pichia pastoris culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane outstanding fourth Pichia pastoris bacterial classification and insert in gnotobasis in the triangular flask that contains the outstanding fourth Pichia pastoris culture medium of sterilizing, at 34 ℃, the rate of shaking 1000rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 4wt% contains the access of triangular flask strain liquid in the fermentation tank of outstanding fourth Pichia pastoris culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the outstanding fourth Pichia pastoris culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain outstanding fourth Pichia yeast liquid;
(2) preparation of koji mould liquid:
A, preparation aspergillus japonicus culture medium raw material, comprise following component and weight percent content: sucrose 2.9%, NaNO30.29%, MgSO47H2O0.0468%, KCl0.0046%, FeSO44H2O0.001%, K2HPO40.095%, agar 1.42%, surplus is distilled water, and pH is 6.0;
The preparation of b, fluid nutrient medium: by Japanese aspergillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus japonicus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus japonicus bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus japonicus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 4wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus japonicus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus japonicus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain koji mould liquid;
(3) preparation of aspergillus terreus liquid:
A, preparation Aspergillus terreus culture medium raw material, comprise following component and weight percent content: sucrose 2.9%, NaNO30.29%, MgSO47H2O0.046%, KCl0.0046%, FeSO44H2O0.001%, K2HPO40.095%, agar 1.42%, surplus is distilled water, and pH is 6.0;
The preparation of b, fluid nutrient medium: press Aspergillus terreus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized Aspergillus terreus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane Aspergillus terreus bacterial classification and insert in gnotobasis in the triangular flask that contains the Aspergillus terreus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 4wt% contains the access of triangular flask strain liquid in the fermentation tank of Aspergillus terreus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the Aspergillus terreus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain aspergillus terreus liquid;
(4) preparation of bacillus cereus bacterium liquid:
A, preparation bacillus cereus culture medium raw material comprise following component and weight percent content: peptone 0.5%, beef leaching thing 0.3%, NaCl0.5%, agar 1.4%, and surplus is distilled water;
The preparation of b, fluid nutrient medium: by bacillus cereus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized bacillus cereus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane bacillus cereus bacterial classification and insert in gnotobasis in the triangular flask that contains the bacillus cereus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 4wt% contains the access of triangular flask strain liquid in the fermentation tank of bacillus cereus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the bacillus cereus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain bacillus cereus bacterium liquid;
(5) preparation of thermophilic liquefaction bacillus bacterium liquid:
A, the thermophilic liquefaction bacillus culture medium raw material of preparation comprise following component and weight percent content: peptone 0.48%, beef leaching thing 0.28%, NaCl0.48%, agar 1.48%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: by thermophilic liquefaction bacillus culture medium formulated raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized thermophilic liquefaction bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get the thermophilic liquefaction in inclined-plane bacillus bacterial classification and insert in gnotobasis in the triangular flask that contains the thermophilic liquefaction bacillus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 4wt% contains the access of triangular flask strain liquid in the fermentation tank of thermophilic liquefaction bacillus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the thermophilic liquefaction bacillus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain thermophilic liquefaction bacillus bacterium liquid;
(6) preparation of stearothermophilus ground bacillus bacterium liquid:
A, preparation stearothermophilus ground bacillus culture medium raw material comprise following component and weight percent content: peptone 0.48%, beef leaching thing 0.28%, NaCl0.48%, agar 1.48%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: press stearothermophilus ground bacillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized stearothermophilus ground bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane stearothermophilus ground bacillus bacterial classification and insert in gnotobasis in the triangular flask that contains the stearothermophilus ground bacillus culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 4wt% contains the access of triangular flask strain liquid in the fermentation tank of stearothermophilus ground bacillus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the stearothermophilus ground bacillus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain stearothermophilus ground bacillus bacterium liquid;
(7) preparation of aspergillus nidulans liquid:
A, preparation aspergillus nidulans culture medium raw material, comprise following component and weight percent content: agar 1.46%, surplus are the 12Brix. brewer's wort;
The preparation of b, fluid nutrient medium: press aspergillus nidulans culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus nidulans culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus nidulans bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus nidulans culture medium of sterilizing, at 34 ℃, the rate of shaking 100rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 4wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus nidulans culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus nidulans culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain aspergillus nidulans liquid;
(8) with outstanding fourth Pichia yeast liquid, koji mould liquid, aspergillus terreus liquid, bacillus cereus bacterium liquid, thermophilic liquefaction bacillus bacterium liquid, stearothermophilus ground bacillus bacterium liquid, aspergillus nidulans liquid is puddled in husk by prescription, adding and keeping humidity behind the water is 40～60%, windrow covers fermentation, when piling temperature greater than 40 ℃, husk is moved to fermentation tank continue fermentation, and ventilation 30min/3h, every 12h turning once, control heap temperature is at 60 ℃ simultaneously, humidity is 40%, shakeout the stockpile cooling behind the fermentation 7d, below the heap temperature drop to 20 ℃, humidity is down to 15%, utilizes swill to build the working environment of the organic wet refuse degraded of simulation for mixed bacterium liquid at last, tame seven kinds of bacterial classifications to the adaptability of organic wet refuse and each other with the suitable natural disposition of indigenous bacterial classification, namely prepare organic wet refuse degraded active bio-compounding agent of dissolving.
Embodiment 3
A kind of organic wet refuse degraded active bio-compounding agent of dissolving, comprise following component and weight percent content: outstanding fourth Pichia yeast liquid 1.0%, koji mould liquid 1.0%, aspergillus terreus liquid 1.0%, bacillus cereus bacterium liquid 1.0%, thermophilic liquefaction bacillus bacterium liquid 1.0%, stearothermophilus ground bacillus bacterium liquid 1.0%, aspergillus nidulans liquid 1.0%, husk 60%, swill 5%, surplus is water.Bacteria containing amount in the above-mentioned bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
The dissolve preparation method of active bio-compounding agent may further comprise the steps the degraded of organic wet refuse:
(1) preparation of outstanding fourth Pichia yeast liquid:
A, the outstanding fourth Pichia pastoris culture medium raw material of preparation comprise following component and weight percent content: agar 1.6%, 5 ° of brewer's worts 98.4%;
The preparation of b, fluid nutrient medium: by outstanding fourth Pichia pastoris culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized outstanding fourth Pichia pastoris culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane outstanding fourth Pichia pastoris bacterial classification and insert in gnotobasis in the triangular flask that contains the outstanding fourth Pichia pastoris culture medium of sterilizing, at 34 ℃, the rate of shaking 200rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 5wt% contains the access of triangular flask strain liquid in the fermentation tank of outstanding fourth Pichia pastoris culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the outstanding fourth Pichia pastoris culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain outstanding fourth Pichia yeast liquid;
(2) preparation of koji mould liquid:
A, preparation aspergillus japonicus culture medium raw material, comprise following component and weight percent content: sucrose 3.0%, NaNO30.3%, MgSO47H2O0.048%, KCl0.0048%, FeSO44H2O0.001%, K2HPO40.096%, agar 1.44%, surplus is distilled water, and pH is 6.5;
The preparation of b, fluid nutrient medium: by Japanese aspergillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus japonicus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus japonicus bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus japonicus culture medium of sterilizing, at 34 ℃, the rate of shaking 200rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 5wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus japonicus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus japonicus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain koji mould liquid;
(3) preparation of aspergillus terreus liquid:
A, preparation Aspergillus terreus culture medium raw material, comprise following component and weight percent content: sucrose 3.0%, NaNO30.3%, MgSO47H2O0.048%, KCl0.0048%, FeSO44H2O0.001%, K2HPO40.096%, agar 1.44%, surplus is distilled water, and pH is 6.5;
The preparation of b, fluid nutrient medium: press Aspergillus terreus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized Aspergillus terreus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane Aspergillus terreus bacterial classification and insert in gnotobasis in the triangular flask that contains the Aspergillus terreus culture medium of sterilizing, at 34 ℃, the rate of shaking 200rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 5wt% contains the access of triangular flask strain liquid in the fermentation tank of Aspergillus terreus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the Aspergillus terreus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain aspergillus terreus liquid;
(4) preparation of bacillus cereus bacterium liquid:
A, preparation bacillus cereus culture medium raw material comprise following component and weight percent content: peptone 0.6%, beef leaching thing 0.35%, NaCl0.6%, agar 1.5%, and surplus is distilled water;
The preparation of b, fluid nutrient medium: by bacillus cereus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized bacillus cereus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane bacillus cereus bacterial classification and insert in gnotobasis in the triangular flask that contains the bacillus cereus culture medium of sterilizing, at 34 ℃, the rate of shaking 100～200rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 5wt% contains the access of triangular flask strain liquid in the fermentation tank of bacillus cereus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the bacillus cereus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain bacillus cereus bacterium liquid;
(5) preparation of thermophilic liquefaction bacillus bacterium liquid:
A, the thermophilic liquefaction bacillus culture medium raw material of preparation comprise following component and weight percent content: peptone 0.5%, beef leaching thing 0.30%, NaCl0.5%, agar 1.50%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: by thermophilic liquefaction bacillus culture medium formulated raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized thermophilic liquefaction bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get the thermophilic liquefaction in inclined-plane bacillus bacterial classification and insert in gnotobasis in the triangular flask that contains the thermophilic liquefaction bacillus culture medium of sterilizing, at 34 ℃, the rate of shaking 200rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 5wt% contains the access of triangular flask strain liquid in the fermentation tank of thermophilic liquefaction bacillus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the thermophilic liquefaction bacillus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain thermophilic liquefaction bacillus bacterium liquid;
(6) preparation of stearothermophilus ground bacillus bacterium liquid:
A, preparation stearothermophilus ground bacillus culture medium raw material comprise following component and weight percent content: peptone 0.5%, beef leaching thing 0.30%, NaCl0.5%, agar 1.50%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: press stearothermophilus ground bacillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized stearothermophilus ground bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane stearothermophilus ground bacillus bacterial classification and insert in gnotobasis in the triangular flask that contains the stearothermophilus ground bacillus culture medium of sterilizing, at 34 ℃, the rate of shaking 200rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 5wt% contains the access of triangular flask strain liquid in the fermentation tank of stearothermophilus ground bacillus culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the stearothermophilus ground bacillus culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain stearothermophilus ground bacillus bacterium liquid;
(7) preparation of aspergillus nidulans liquid:
A, preparation aspergillus nidulans culture medium raw material, comprise following component and weight percent content: agar 1.48%, surplus are the 12Brix. brewer's wort;
The preparation of b, fluid nutrient medium: press aspergillus nidulans culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus nidulans culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus nidulans bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus nidulans culture medium of sterilizing, at 34 ℃, the rate of shaking 200rpm cultivates 48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 5wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus nidulans culture medium, at 35 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus nidulans culture medium that above-mentioned condition is added the fermentation tank volume 48h that ferments again, obtain aspergillus nidulans liquid;
(8) with outstanding fourth Pichia yeast liquid, koji mould liquid, aspergillus terreus liquid, bacillus cereus bacterium liquid, thermophilic liquefaction bacillus bacterium liquid, stearothermophilus ground bacillus bacterium liquid, aspergillus nidulans liquid is puddled in husk by prescription, adding and keeping humidity behind the water is 60%, windrow covers fermentation, when piling temperature greater than 40 ℃, husk is moved to fermentation tank continue fermentation, and ventilation 30min/3h, every 12h turning once, control heap temperature is at 70 ℃ simultaneously, humidity is 50%, shakeout the stockpile cooling behind the fermentation 7d, below the heap temperature drop to 20 ℃, humidity is down to 15%, utilizes swill to build the working environment of the organic wet refuse degraded of simulation for mixed bacterium liquid at last, tame seven kinds of bacterial classifications to the adaptability of organic wet refuse and each other with the suitable natural disposition of indigenous bacterial classification, namely prepare organic wet refuse degraded active bio-compounding agent of dissolving.
This bio-compounding agent is used for garbage degradation, the processing time be on March 22nd, 2012 to April 4, result of the test is as follows.
The organic wet refuse degraded active bio-compounding agent log of dissolving
Test site: go up marine mountain hospital hepatitis ward test period: 9～28 October in 1996
Claims (9)
1. organic wet refuse degraded active bio-compounding agent of dissolving, it is characterized in that, this complexing agent comprises following component and weight percent content: outstanding fourth Pichia yeast liquid 0.6～1.0%, koji mould liquid 0.6～1.0%, aspergillus terreus liquid 0.6～1.0%, bacillus cereus bacterium liquid 0.6～1.0%, thermophilic liquefaction bacillus bacterium liquid 0.6～1.0%, stearothermophilus ground bacillus bacterium liquid 0.6～1.0%, aspergillus nidulans liquid 0.6～1.0%, husk 50～60%, swill 4～5%, surplus is water.
2. a kind of organic wet refuse degraded according to claim 1 active bio-compounding agent of dissolving is characterized in that the bacteria containing amount of described outstanding fourth Pichia yeast liquid is 1 * 10
8～9 * 10
12Individual/ml.
3. a kind of organic wet refuse degraded according to claim 1 active bio-compounding agent of dissolving is characterized in that the bacteria containing amount of described koji mould liquid is 1 * 10
8～9 * 10
12Individual/ml.
4. a kind of organic wet refuse degraded according to claim 1 active bio-compounding agent of dissolving is characterized in that the bacteria containing amount of described aspergillus terreus liquid is 1 * 10
8～9 * 10
12Individual/ml.
5. a kind of organic wet refuse degraded according to claim 1 active bio-compounding agent of dissolving is characterized in that the bacteria containing amount of described bacillus cereus bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml
6. a kind of organic wet refuse degraded according to claim 1 active bio-compounding agent of dissolving is characterized in that the bacteria containing amount of described thermophilic liquefaction bacillus bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
7. a kind of organic wet refuse degraded according to claim 1 active bio-compounding agent of dissolving is characterized in that the bacteria containing amount of described stearothermophilus ground bacillus bacterium liquid is 1 * 10
8～9 * 10
12Individual/ml.
8. a kind of organic wet refuse degraded according to claim 1 active bio-compounding agent of dissolving is characterized in that the bacteria containing amount of described aspergillus nidulans liquid is 1 * 10
8～9 * 10
12Individual/ml.
9. according to the dissolve preparation method of active bio-compounding agent of each described organic wet refuse degraded among the claim 1-8, it is characterized in that this method may further comprise the steps:
(1) preparation of outstanding fourth Pichia yeast liquid:
A, the outstanding fourth Pichia pastoris culture medium raw material of preparation comprise following component and weight percent content: agar 1.4～1.6%, 5 ° of brewer's worts 98.4～98.6%;
The preparation of b, fluid nutrient medium: by outstanding fourth Pichia pastoris culture medium prescription preparation raw material, at 015MPa, 121 ℃, sterilization 15min obtains sterilized outstanding fourth Pichia pastoris culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane outstanding fourth Pichia pastoris bacterial classification and insert in gnotobasis in the triangular flask that contains the outstanding fourth Pichia pastoris culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of outstanding fourth Pichia pastoris culture medium, at 30～45 ℃, ventilating ratio is 1: 1～2 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the outstanding fourth Pichia pastoris culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain outstanding fourth Pichia yeast liquid;
(2) preparation of koji mould liquid:
A, preparation aspergillus japonicus culture medium raw material, comprise following component and weight percent content: sucrose 2.8～3.0%, NaNO30.28～0.3%, MgSO47H2O0.045～0.048%, KCl0.0045～0.0048%, FeSO44H2O0.001%, K2HPO40.095～0.096%, agar 1.4%～1.44%, surplus is distilled water, and pH is 6.0～6.5;
The preparation of b, fluid nutrient medium: by Japanese aspergillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus japonicus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus japonicus bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus japonicus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus japonicus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 48h, and then by 1/3 the aspergillus japonicus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain koji mould liquid;
(3) preparation of aspergillus terreus liquid:
A, preparation Aspergillus terreus culture medium raw material, comprise following component and weight percent content: sucrose 2.8～3.0%, NaNO30.28～0.3%, MgSO47H2O0.045～0.048%, KCl0.0045～0.0048%, FeSO44H2O0.001%, K2HPO40.095～0.096%, agar 1.4%～1.44%, surplus is distilled water, and pH is 6.0～6.5;
The preparation of b, fluid nutrient medium: press Aspergillus terreus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized Aspergillus terreus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane Aspergillus terreus bacterial classification and insert in gnotobasis in the triangular flask that contains the Aspergillus terreus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of Aspergillus terreus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the Aspergillus terreus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain aspergillus terreus liquid;
(4) preparation of bacillus cereus bacterium liquid:
A, preparation bacillus cereus culture medium raw material comprise following component and weight percent content: peptone 0.4～0.6%, beef leaching thing 0.25～0.35%, NaCl0.4～0.6%, agar 1.4～1.5%, and surplus is distilled water;
The preparation of b, fluid nutrient medium: by bacillus cereus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized bacillus cereus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane bacillus cereus bacterial classification and insert in gnotobasis in the triangular flask that contains the bacillus cereus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of bacillus cereus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the bacillus cereus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain bacillus cereus bacterium liquid;
(5) preparation of thermophilic liquefaction bacillus bacterium liquid:
A, the thermophilic liquefaction bacillus culture medium raw material of preparation, comprise following component and weight percent content: peptone 0.45～0.5%, beef leaching thing 0.27～0.30%, NaCl0.45～0.5%, agar 1.45～1.50%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: by thermophilic liquefaction bacillus culture medium formulated raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized thermophilic liquefaction bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get the thermophilic liquefaction in inclined-plane bacillus bacterial classification and in gnotobasis, insert in the triangular flask that contains the thermophilic liquefaction bacillus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of thermophilic liquefaction bacillus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the thermophilic liquefaction bacillus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain thermophilic liquefaction bacillus bacterium liquid;
(6) preparation of stearothermophilus ground bacillus bacterium liquid:
A, preparation stearothermophilus ground bacillus culture medium raw material, comprise following component and weight percent content: peptone 0.45～0.5%, beef leaching thing 0.27～0.30%, NaCl0.45～0.5%, agar 1.45～1.50%, surplus are distilled water, and pH is 7.0;
The preparation of b, fluid nutrient medium: press stearothermophilus ground bacillus culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized stearothermophilus ground bacillus culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane stearothermophilus ground bacillus bacterial classification and in gnotobasis, insert in the triangular flask that contains the stearothermophilus ground bacillus culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of stearothermophilus ground bacillus culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the stearothermophilus ground bacillus culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain stearothermophilus ground bacillus bacterium liquid;
(7) preparation of aspergillus nidulans liquid:
A, preparation aspergillus nidulans culture medium raw material, comprise following component and weight percent content: agar 1.45～1.48%, surplus are the 12Brix. brewer's wort;
The preparation of b, fluid nutrient medium: press aspergillus nidulans culture medium prescription preparation raw material, at 0.15MPa, 121 ℃, sterilization 15min obtains sterilized aspergillus nidulans culture medium;
The preparation of c, triangular flask bacterial classification: get inclined-plane aspergillus nidulans bacterial classification and insert in gnotobasis in the triangular flask that contains the aspergillus nidulans culture medium of sterilizing, at 30～45 ℃, the rate of shaking 100～200rpm cultivates 24～48h, obtains the triangular flask strain liquid;
D, the preparation of producing bacterial classification: the inoculum concentration by 3wt%～5wt% contains the access of triangular flask strain liquid in the fermentation tank of aspergillus nidulans culture medium, at 30～45 ℃, ventilating ratio is 1: 1 (V/V), liquid amount is 1/3 of fermentation tank volume, fermentation 24～48h, and then by 1/3 the aspergillus nidulans culture medium that above-mentioned condition is added the fermentation tank volume, the 24～48h that ferments again, obtain aspergillus nidulans liquid;
(8) with outstanding fourth Pichia yeast liquid, koji mould liquid, aspergillus terreus liquid, bacillus cereus bacterium liquid, thermophilic liquefaction bacillus bacterium liquid, stearothermophilus ground bacillus bacterium liquid, aspergillus nidulans liquid is puddled in husk by prescription, adding and keeping humidity behind the water is 40～60%, windrow covers fermentation, when piling temperature between 40～60 ℃, husk is moved to fermentation tank continue fermentation, and the 30～60min/3h that ventilates, every 12h turning once, control heap temperature is at 50～70 ℃ simultaneously, humidity is 20～50%, shakeout the stockpile cooling behind fermentation 7～10d, below the heap temperature drop to 20 ℃, humidity is down to 20～10%, utilize swill to build the working environment of the organic wet refuse degraded of simulation for mixed bacterium liquid at last, tame seven kinds of bacterial classifications to the adaptability of organic wet refuse and each other with the suitable natural disposition of indigenous bacterial classification, namely prepare organic wet refuse degraded active bio-compounding agent of dissolving.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310180290.8A CN103272361B (en) | 2013-05-15 | 2013-05-15 | A kind of organic wet refuse degraded is dissolved active bio-compounding agent and preparation method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104531550A (en) * | 2014-11-14 | 2015-04-22 | 山东城矿环保集团有限公司 | High-temperature kitchen waste degradation microbial agent and preparing and using method thereof |
| CN107541469A (en) * | 2017-10-11 | 2018-01-05 | 清水源（上海）环保科技有限公司 | A kind of compound bio digestion agent and preparation method thereof and its application |
| CN108998388A (en) * | 2018-07-18 | 2018-12-14 | 梅金琪 | A kind of preparation method of waste disposal agent |
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| US20070122874A1 (en) * | 2005-11-25 | 2007-05-31 | Rangasamy Suthanthararajan | Novel microbial consortium and use thereof for liquefaction of solid organic matter |
| CN102531766A (en) * | 2011-09-15 | 2012-07-04 | 北京世纪阿姆斯生物技术股份有限公司 | Microbial decomposing agent and production method thereof |
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| US20060030023A1 (en) * | 2004-08-03 | 2006-02-09 | Somers Heather K | Bio-waste enzyme packet |
| US20070122874A1 (en) * | 2005-11-25 | 2007-05-31 | Rangasamy Suthanthararajan | Novel microbial consortium and use thereof for liquefaction of solid organic matter |
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| CN104531550A (en) * | 2014-11-14 | 2015-04-22 | 山东城矿环保集团有限公司 | High-temperature kitchen waste degradation microbial agent and preparing and using method thereof |
| CN107541469A (en) * | 2017-10-11 | 2018-01-05 | 清水源（上海）环保科技有限公司 | A kind of compound bio digestion agent and preparation method thereof and its application |
| CN107541469B (en) * | 2017-10-11 | 2020-07-03 | 清水源（上海）环保科技有限公司 | Composite biological digestion agent and preparation method and application thereof |
| CN108998388A (en) * | 2018-07-18 | 2018-12-14 | 梅金琪 | A kind of preparation method of waste disposal agent |
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| CN103272361B (en) | 2015-11-04 |
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