CN113652371A - Ultrahigh-temperature kitchen waste treatment microbial inoculum and preparation method thereof - Google Patents
Ultrahigh-temperature kitchen waste treatment microbial inoculum and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of microbial preparations, and particularly relates to an ultrahigh-temperature kitchen waste treatment microbial inoculum and a preparation method thereof. Collecting high-temperature hot springs, deep-sea hot liquid and sludge composting system samples at the fermentation stage of 70-80 ℃; respectively carrying out microbial enrichment on the samples by adopting an NB culture medium, mixing the enrichments, and then carrying out continuous culture by adopting the NB culture medium to obtain ultrahigh-temperature composite microbial fermentation liquor; drying humic acid modified kaolin, starch and wheat bran, grinding and sieving to obtain microbial inoculum carrier powder; adding the skim milk protective agent into the ultrahigh-temperature composite microorganism fermentation liquor, stirring to obtain a composite bacterial liquid, adding the bacterial agent carrier powder, stirring and mixing, performing suction filtration to obtain filter residues, and drying, grinding and sieving the filter residues to obtain the ultrahigh-temperature kitchen waste treatment bacterial agent. The survival rate of the microbial strains in the microbial inoculum is high, the storage time is long, and a technical thought is provided for the storage and transportation of the ultra-high temperature microbial inoculum.
Description
Technical Field
The invention belongs to the technical field of microbial preparations, and particularly relates to an ultrahigh-temperature kitchen waste treatment microbial inoculum and a preparation method thereof.
Background
The pollution caused by the kitchen waste becomes the main problem of urban environment pollution, the kitchen waste refers to food residues generated in food processing engineering and food residues after eating and drinking consumption, mainly comprises organic substances such as starch, cellulose, protein and the like, and a large amount of malodorous gas is easily generated under the action of harmful bacteria to influence the normal life of people.
In recent years, biological treatment methods and non-biological treatment methods are mainly adopted for treating kitchen waste, wherein the non-biological treatment methods mainly comprise incineration, dehydration treatment and vacuum frying treatment, the biological treatment methods mainly utilize microorganisms to ferment and convert organic substances to obtain inorganic substances, but the existing microbial treatment methods have low treatment efficiency.
For an extreme thermophilic microorganism liquid microbial inoculum, the activity of strains is easily influenced when the liquid microbial inoculum is stored at room temperature for a long time, so that part of strains die, and the probability of infecting mixed bacteria is increased, therefore, a method for preparing the microbial inoculum capable of improving the survival rate and the storage time of extreme thermophilic microorganism strains is developed.
Disclosure of Invention
Aiming at the problems in the prior art, the invention enriches the microorganisms of the sludge composting system samples in the high-temperature hot spring, deep-sea hot liquid and high-temperature fermentation stages, obtains the compound fermentation liquor by amplification culture, and obtains the solid microbial inoculum with high strain survival rate, long storage time and high treatment efficiency by adsorbing the microorganisms in the compound fermentation liquor by a specific microbial inoculum carrier.
In order to realize the aim, the invention provides a preparation method of an ultrahigh-temperature kitchen waste treatment microbial inoculum, which specifically comprises the following steps:
collecting high-temperature hot springs, deep-sea hot liquid and sludge composting system samples at the fermentation stage of 70-80 ℃; respectively carrying out microbial enrichment on the samples by adopting NB culture media to respectively obtain high-temperature hot spring microbial enrichment, deep sea hot liquid microbial enrichment and high-temperature sludge fermentation microbial enrichment;
mixing the high-temperature hot spring microorganism enrichment, the deep sea hot liquid microorganism enrichment and the high-temperature sludge fermentation microorganism enrichment, and then continuously culturing by adopting an NB (nuclear magnetic resonance) culture medium to obtain ultrahigh-temperature composite microorganism fermentation liquor;
drying humic acid modified kaolin, starch and wheat bran, grinding and sieving to obtain microbial inoculum carrier powder;
adding the skim milk protective agent into the ultrahigh-temperature composite microorganism fermentation liquor, stirring to obtain a composite bacterial liquid, adding the bacterial agent carrier powder, stirring and mixing, performing suction filtration to obtain filter residues, and drying, grinding and sieving the filter residues to obtain the ultrahigh-temperature kitchen waste treatment bacterial agent.
Further, the NB medium is: 10.0g of peptone, 3.0g of beef extract powder and 5.0g of NaCl5, adding deionized water to the mixture until the volume is 1000mL, and keeping the pH value natural.
Further, the temperature of the enrichment culture process is 70-80 ℃; the continuous culture process is carried out at 70-80 ℃ and 160-180rpm for 90-120 h.
Further, the compound bacterial liquid and the microbial inoculum carrier are mixed according to the solid-liquid ratio of 1: 1.5-2.
Based on the same invention concept, the embodiment of the invention also provides an ultrahigh-temperature kitchen waste treatment microbial inoculum, wherein the ultrahigh-temperature kitchen waste treatment microbial inoculum is prepared by the preparation method;
the microbial inoculum comprises the following microorganisms in percentage by weight: 40-50% of thermophilic bacteria; 10-20% of staphylococcus; 10-20% of Planococcus; 2-10% of hyperthermophile; 2-10% of bacillus; 2-5% of true silk fungus.
Further, the effective viable count of the microbial inoculum is 6.5-10 multiplied by 108CFU/g。
Has the advantages that:
(1) according to the invention, high-temperature hot springs, deep-sea hydrothermal solution and sludge composting system samples in a high-temperature fermentation stage are subjected to microorganism enrichment, and a compound microbial agent is obtained after continuous culture, wherein the compound microbial agent has more microorganism types which are all high-temperature stable microorganisms, and can be fermented by using organic matters in kitchen garbage under a high-temperature condition to rapidly degrade the organic matters. The activity of the hyperthermophiles and the thermophilic bacteria is higher in the high-temperature stage, and a large amount of amylase, protease, cellulase and the like with high thermal stability can be generated to decompose organic matters; the staphylococcus can decompose small molecular organic matters such as glucose, maltose, sucrose and the like; the Planococcus has strong capability of decomposing protein; the bacillus can secrete lipase with strong activity; the genus true silk has strong degradation ability to cellulose and lignin. The types of enzymes and metabolized organic matters generated among different microorganisms are different, and the microorganisms participate in fermentation simultaneously to have a synergistic effect, so that organic matters in the kitchen waste are rapidly degraded.
(2) According to the invention, the humic acid kaolin, the starch and the wheat bran are used as microorganism carriers, the microorganism is protected by the protective agent, and the two are mixed and stirred at high temperature, so that the microorganism carriers adsorb the microorganism to obtain the solid microorganism fungicide, the survival rate of microorganism strains in the fungicide is high, the storage time is long, and a technical thought is provided for the storage and transportation of the ultra-high temperature microorganism fungicide.
Detailed Description
In order to make the technical problems, technical solutions and advantages of the present invention more apparent, the following detailed description is given with reference to specific embodiments, but the scope of the present invention is not limited to the following specific embodiments.
Unless otherwise defined, all terms of art used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by existing methods.
In the embodiment of the invention, the high-temperature hot spring is collected from Yunnan Tengchong (98 degrees 05-98 degrees 45'E, 24 degrees 38-25 degrees 52' N), the deep-sea hot liquid is collected from Okinawa sea troughs (126 degrees 53.84'E, 27 degrees 47.44' N, deep 966.9 meters), and the sludge composting system is collected from an ultra-high temperature composting plant (116 degrees 56.21'E, 40 degrees 3.1' N) in the cisterm of Beijing.
In an embodiment of the invention, the NB (nutrient broth) medium is: 10.0g of peptone, 3.0g of beef extract powder and 5.0g of NaCl, adding deionized water to the mixture until the volume is 1000mL, and keeping the pH value natural.
Example 1
An ultrahigh-temperature kitchen waste treatment microbial inoculum is prepared by the following method:
collecting samples of a sludge composting system in a high-temperature hot spring, deep sea hot liquid and 75 ℃ fermentation stage; respectively carrying out microbial enrichment on the samples by adopting NB culture media to respectively obtain high-temperature hot spring microbial enrichment, deep sea hot liquid microbial enrichment and high-temperature sludge fermentation microbial enrichment;
mixing the high-temperature hot spring microorganism concentrate, the deep sea hot liquid microorganism concentrate and the high-temperature sludge fermentation microorganism concentrate according to the volume ratio of 1:1:1, and then continuously culturing by adopting an NB culture medium to obtain ultrahigh-temperature compound microorganism fermentation liquor;
drying humic acid modified kaolin, starch and wheat bran at 105 ℃, grinding and sieving with a 100-mesh sieve to obtain microbial inoculum carrier powder;
adding a skim milk protective agent into the ultrahigh-temperature composite microorganism fermentation liquor by 1% (w/v), stirring for 2h to obtain a composite bacterial liquid, adding the bacterial agent carrier powder, stirring and mixing, performing suction filtration to obtain filter residues, drying and grinding the filter residues at 75 ℃ and sieving with a 100-mesh sieve to obtain the ultrahigh-temperature kitchen waste treatment bacterial agent.
The effective viable count of the microbial inoculum is 6.5-10 multiplied by 108CFU/g, microbial composition: 50% of thermophilic bacteria; staphylococcus 15%, Planococcus 15%; hyperthermophiles belong to 8 percent; 10% of bacillus; the genus of true silk is 2%.
Example 2
An ultrahigh-temperature kitchen waste treatment microbial inoculum is prepared by the following method:
collecting samples of a sludge composting system in a high-temperature hot spring, deep sea hot liquid and 80 ℃ fermentation stage; respectively carrying out microbial enrichment on the samples by adopting NB culture media to respectively obtain high-temperature hot spring microbial enrichment, deep sea hot liquid microbial enrichment and high-temperature sludge fermentation microbial enrichment;
mixing the high-temperature hot spring microorganism concentrate, the deep sea hot liquid microorganism concentrate and the high-temperature sludge fermentation microorganism concentrate according to the volume ratio of 2:1:1, and then continuously culturing by adopting an NB culture medium to obtain ultrahigh-temperature compound microorganism fermentation liquor;
drying humic acid modified kaolin, starch and wheat bran at 110 ℃, grinding and sieving with a 100-mesh sieve to obtain microbial inoculum carrier powder;
adding a skim milk protective agent into the ultrahigh-temperature composite microorganism fermentation liquor by 1% (w/v), stirring for 2h to obtain a composite bacterial liquid, adding the bacterial agent carrier powder, stirring and mixing, performing suction filtration to obtain filter residues, drying and grinding the filter residues at 70 ℃ and sieving with a 100-mesh sieve to obtain the ultrahigh-temperature kitchen waste treatment bacterial agent.
The effective viable count of the microbial inoculum is 6.5-10 multiplied by 108CFU/g, microbial composition: 45% of thermophilic bacteria; staphylococcus 20%, Planococcus 15%; hyperthermophiles 5%; 10% of bacillus; the genus of true silk is 5%.
Example 3
An ultrahigh-temperature kitchen waste treatment microbial inoculum is prepared by the following method:
collecting samples of a sludge composting system in a high-temperature hot spring, deep sea hot liquid and 70 ℃ fermentation stage; respectively carrying out microbial enrichment on the samples by adopting NB culture media to respectively obtain high-temperature hot spring microbial enrichment, deep sea hot liquid microbial enrichment and high-temperature sludge fermentation microbial enrichment;
mixing the high-temperature hot spring microorganism concentrate, the deep sea hot liquid microorganism concentrate and the high-temperature sludge fermentation microorganism concentrate according to the volume ratio of 1:1:2, and then continuously culturing by adopting an NB culture medium to obtain ultrahigh-temperature compound microorganism fermentation liquor;
drying humic acid modified kaolin, starch and wheat bran at 100 ℃, grinding and sieving with a 100-mesh sieve to obtain microbial inoculum carrier powder;
adding a skim milk protective agent into the ultrahigh-temperature composite microorganism fermentation liquor by 1% (w/v), stirring for 2h to obtain a composite bacterial liquid, adding the bacterial agent carrier powder, stirring and mixing, performing suction filtration to obtain filter residues, drying and grinding the filter residues at 80 ℃ and sieving with a 100-mesh sieve to obtain the ultrahigh-temperature kitchen waste treatment bacterial agent.
The effective viable count of the microbial inoculum is 6.5-10 multiplied by 108CFU/g, microbial composition: 50% of thermophilic bacteria; staphylococcus 10%, Planococcus 20%; hyperthermophiles 5%; 10% of bacillus; the genus of true silk is 5%.
Comparative example 1
And (3) carrying out microorganism enrichment by taking a sludge composting system in a high-temperature hot spring and 70 ℃ fermentation stage as a sample, and obtaining the treating microbial inoculum by the same other steps as the example 1. The effective viable count of the microbial inoculum is 8.0 multiplied by 108CFU/g, microbial composition: the microbial composition is as follows: 46% of the genus thermophilus; staphylococcus 18%, Planococcus 14%; hyperthermophiles 5%; 15% of bacillus; the genus of true silk is 2%.
Comparative example 2
The deep sea hydrothermal solution and the sludge composting system at the 70 ℃ fermentation stage are used as samples for microorganism enrichment, and other steps are the same as those in the example 1 to obtain the treatment microbial inoculum. The effective viable count of the microbial inoculum is 8.0 multiplied by 108CFU/g, microbial composition: microbial compositionComprises the following steps: 50% of thermophilic bacteria; staphylococcus 15%, Planococcus 15%; hyperthermophiles 5%; 10% of bacillus; the genus of true silk is 5%.
Comparative example 3
And (3) carrying out microorganism enrichment by taking deep sea hydrothermal solution and high temperature hot spring as samples, and obtaining the treating microbial inoculum by the same other steps as the example 1. The effective viable count of the microbial inoculum is 8.0 multiplied by 108CFU/g, microbial composition: the microbial composition is as follows: 45% of thermophilic bacteria; staphylococcus 10%, Planococcus 20%; hyperthermophiles 5%; 15% of bacillus; the genus of true silk is 5%.
Application example
The kitchen waste raw materials collected from restaurants, dining halls and other places are screened out of impurities such as plastics, paper towels, cullet and the like in the kitchen waste raw materials by methods such as vibration screening, drum screening and the like, and the undersize products are crushed and further subjected to impurity removal to obtain the kitchen waste with high water content; and then removing a part of water, oil and salt from the kitchen waste through a squeezing device to obtain the kitchen waste with the water content of about 70 wt%.
Feeding the kitchen waste after primary treatment into a fermentation bin, fully and uniformly mixing the kitchen waste with one or more auxiliary materials (the water content is 5-10%) of sawdust, rice husk, wheat bran or wood shavings by mechanical stirring to adjust the water content of the whole material to 50-60%, then adding an ultrahigh-temperature kitchen waste treatment microbial inoculum according to the ratio of the microbial inoculum to the material (1: 100), and then uniformly mixing the material and the microbial inoculum by mechanical stirring. The temperature in the fermentation bin is maintained at 65-75 ℃ by electromagnetic heating of the outer layer of the fermentation bin, the motor is arranged to stir positively and negatively for 2 minutes at intervals of 10 minutes and then stir positively and negatively for 2 minutes for repeated circulation to keep the material in an aerobic state all the time, and the oxygen content of the material is detected by using the dissolved oxygen probe. And (5) sampling at regular time, and measuring the real-time moisture content of the material by adopting a rapid moisture content instrument.
TABLE 1 initial fermentation Material Properties
Table 2 change in moisture content of material during fermentation in example 1 and comparative example 1
After high-temperature fermentation for 38 hours, the water content of the material in the comparative example 1 is reduced from 57.00 percent to 24.54 percent; in the embodiment 1, under the action of the ultrahigh-temperature kitchen waste treatment microbial inoculum, the water content of the material is reduced from 57.00% to 6.02%; the result shows that the reduction effect of the embodiment added with the ultrahigh-temperature kitchen waste treatment microbial inoculum is obvious.
The above-mentioned embodiments are only preferred embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered as the technical scope of the present invention, and equivalents and modifications of the technical solutions and concepts of the present invention should be covered by the scope of the present invention.
Claims (6)
1. A preparation method of an ultrahigh-temperature kitchen waste treatment microbial inoculum is characterized by specifically comprising the following steps:
collecting high-temperature hot springs, deep-sea hot liquid and sludge composting system samples at the fermentation stage of 70-80 ℃; respectively carrying out microbial enrichment on the samples by adopting NB culture media to respectively obtain high-temperature hot spring microbial enrichment, deep sea hot liquid microbial enrichment and high-temperature sludge fermentation microbial enrichment;
mixing the high-temperature hot spring microorganism enrichment, the deep sea hot liquid microorganism enrichment and the high-temperature sludge fermentation microorganism enrichment, and then continuously culturing by adopting an NB (nuclear magnetic resonance) culture medium to obtain ultrahigh-temperature composite microorganism fermentation liquor;
drying humic acid modified kaolin, starch and wheat bran, grinding and sieving to obtain microbial inoculum carrier powder;
adding the skim milk protective agent into the ultrahigh-temperature composite microorganism fermentation liquor, stirring to obtain a composite bacterial liquid, adding the bacterial agent carrier powder, stirring and mixing, performing suction filtration to obtain filter residues, and drying, grinding and sieving the filter residues to obtain the ultrahigh-temperature kitchen waste treatment bacterial agent.
2. The preparation method of the ultra-high temperature kitchen waste treatment microbial inoculum according to claim 1, wherein the NB culture medium is: 10.0g of peptone, 3.0g of beef extract powder and 5.0g of NaCl, adding deionized water to the mixture until the volume is 1000mL, and keeping the pH value natural.
3. The preparation method of the ultra-high temperature kitchen waste treatment microbial inoculum according to claim 1, wherein the temperature of the enrichment culture process is 70-80 ℃; the continuous culture process is carried out at 70-80 ℃ and 160-180rpm for 90-120 h.
4. The preparation method of the ultra-high temperature kitchen waste treatment microbial inoculum according to claim 1, wherein the compound bacterial liquid and the microbial inoculum carrier are mixed according to a solid-to-liquid ratio of 1: 1.5-2.
5. An ultra-high temperature kitchen waste treatment microbial inoculum is characterized in that the ultra-high temperature kitchen waste treatment microbial inoculum is prepared by the preparation method of any claim 1 to 4;
the microbial inoculum comprises the following microorganisms in percentage by weight: 40-50% of thermophilic bacteria; 10-20% of staphylococcus; 10-20% of Planococcus; 2-10% of hyperthermophile; 2-10% of bacillus; 2-5% of true silk fungus.
6. The ultra-high temperature kitchen waste treatment microbial inoculum according to claim 5, wherein the effective viable count of the microbial inoculum is 6.5-10 x 108CFU/g。
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