CN112481152A - Microbial agent-enzyme preparation compound and method for degrading kitchen garbage by using same - Google Patents

Microbial agent-enzyme preparation compound and method for degrading kitchen garbage by using same Download PDF

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CN112481152A
CN112481152A CN202011223847.8A CN202011223847A CN112481152A CN 112481152 A CN112481152 A CN 112481152A CN 202011223847 A CN202011223847 A CN 202011223847A CN 112481152 A CN112481152 A CN 112481152A
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microbial agent
enzyme preparation
kitchen waste
enzyme
degrading
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徐毅
倪晶霞
马宝娣
吴小梅
刘胜利
丁慧贤
王自稳
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Shanghai Institute of Technology
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
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    • C12N9/2405—Glucanases
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    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
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Abstract

The invention relates to a microbial agent-enzyme preparation compound and a method for degrading kitchen waste by using the same, wherein the microbial agent-enzyme preparation compound for degrading the kitchen waste comprises a microbial agent and an enzyme preparation, the microbial agent comprises mixed microorganisms of bacillus and wood dust, the enzyme preparation comprises amylase, cellulase, protease and lipase, the mixed microbial agent is prepared by fermenting the mixed microbial strains, and the mixed microbial agent and free enzyme or immobilized enzyme are combined to degrade the kitchen waste. Compared with the prior art, the microbial agent and enzyme preparation compound system provided by the invention can obviously improve the degradation speed and the degradation rate of kitchen waste compared with the microbial agent only used under the same condition, the reduction rate of the kitchen waste reaches more than 94%, and the kitchen waste can be repeatedly degraded for multiple batches, so that the microbial agent and enzyme preparation compound system has a higher application prospect.

Description

Microbial agent-enzyme preparation compound and method for degrading kitchen garbage by using same
Technical Field
The invention relates to the field of kitchen waste degradation, in particular to a microbial agent-enzyme preparation compound for degrading kitchen waste and a preparation method and application thereof.
Background
The organic garbage generally comprises agricultural and forestry and cultivation wastes such as straw leaves, shell sawdust, human and animal excrement and the like, and also comprises production and domestic garbage such as kitchen waste, food waste, sewer sludge and the like. Among various types of municipal waste, organic waste is more difficult to treat than general waste because of its wide source, high yield, complex composition and often various pathogens. Taking food waste as an example, the food waste includes town garbage such as garbage in town vegetable yards, garbage in restaurant food industry, garbage in household kitchens, and melon and fruit peels in various markets, and can also be called as food waste. The kitchen garbage is easy to ferment, deteriorate and rot, so that a large amount of toxins are generated, malodorous gases are emitted, water and atmosphere are polluted, environmental sanitation is seriously affected, and potential threats are formed to the health of people. Therefore, the swill refuse disposal problem is serious before people.
The traditional kitchen waste treatment mode in China is to directly feed the kitchen waste to a farm for feeding pigs, and the treatment mode has multiple potential hazards to the environment and is very easy to cause disease propagation. In recent years, avian influenza, foot-and-mouth disease and other infectious diseases are prevalent in a large scale, and cross infection among people and livestock has already made a great threat to human health and social stability. At present, relevant policies are formulated in succession in Shanghai, Suzhou and other markets, and the hogwash is prohibited from being used for feeding pigs in the open.
The present treatment method of urban domestic refuse mainly includes sanitary landfill, incineration and composting. And each of the three methods has advantages and disadvantages. Sanitary landfills have the advantage of high throughput and low overall cost, and have the disadvantage of high land occupation and percolation problems to be considered. The burning can reduce the amount in a short time, recover energy and kill various viruses and bacteria, but the initial investment is large, the operation cost is high and the atmospheric pollution is also caused. The compost has the advantages of high recycling degree, but the garbage components are complex, the process conditions are difficult to control, and the offensive odor is easily generated. The municipal solid waste is mainly treated by landfill in China, and accounts for more than 70% of the total treatment capacity; secondly, high-temperature composting which accounts for more than 20 percent; the burning amount is very small. Therefore, how to improve the disposal mode of domestic garbage, especially organic garbage, is an important step for improving the environmental quality.
The general kitchen waste is rich in resources, contains a large amount of carbohydrate, protein, fat and the like, and is not beneficial to direct utilization. But has a high degree of biodegradability. At present, the degradation processor of kitchen garbage is common in Japan, such as the swill processor disclosed in patent CN 2894847Y. The research on the degradation of the kitchen waste is successful, and China also introduces some degradation machines, but the core microorganisms for degrading the kitchen waste are mainly imported.
In recent years, some enterprises in China independently develop microbial strains and methods for treating kitchen waste, such as the screening of kitchen waste degrading strains disclosed in CN107760616A and the degradation method disclosed in patent CN102952759A, but the degradation effect is not good, the content of grease in the kitchen waste is high, effective degradation cannot be achieved, the degradation time is long, and the degradation rate is low.
In order to improve the degradation efficiency, a new degradation mode of the microbial agent is needed. The early reports that the mixed microbial strains are used for degrading the kitchen garbage only have slow degradation speed, long period and low degradation rate.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a microbial agent-enzyme preparation compound for degrading kitchen waste and a preparation method and application thereof.
The purpose of the invention can be realized by the following technical scheme:
the microbial agent-enzyme preparation compound for degrading kitchen waste comprises a microbial agent and an enzyme preparation, wherein the microbial agent comprises mixed microorganisms of bacillus and wood dust, and the enzyme preparation comprises amylase, cellulase, protease and lipase.
Further, the mass ratio of the microbial agent to the enzyme preparation is 20: 2-25: 2.
Further, the microbial agent comprises mixed microorganisms of bacillus and wood chips in a mass ratio of 4-6: 1.
Further, the mass ratio of the amylase to the cellulase to the protease to the lipase is as follows: 0.8-1.1: 0.8-1.1: 0.8-1.1: 0.8 to 1.1.
Further, the bacillus comprises bacillus licheniformis and bacillus subtilis in a mass ratio of 1: 2.
Further, the bacillus subtilis is preserved by China general microbiological culture Collection center with the preservation number (CGMCC1.14985, CGMCC1.7417), and the bacillus licheniformis is preserved by China general microbiological culture Collection center with the preservation number (CGMCC 1.813).
The preparation method of the microbial agent-enzyme preparation compound comprises the following steps: and (2) fermenting and culturing the compound microbial inoculum in a culture medium for 24-36 hours at the fermentation temperature of 37 ℃, mixing the compound microbial inoculum with wood dust, air-drying to obtain a microbial inoculum, and adding an enzyme preparation to obtain a microbial inoculum-enzyme preparation compound.
Further, the culture medium is a nutrient broth culture medium.
Further, the enzyme preparation is free enzyme or immobilized enzyme.
Further, when the enzyme preparation is immobilized enzyme, the enzyme preparation is self-adsorbed to wood chips, and then glutaraldehyde is added for crosslinking.
The application of the microbial agent-enzyme preparation compound in degrading kitchen garbage comprises the following steps: the kitchen waste is poured into a degradation container, the microbial agent and enzyme complex is added, water is added to enable the water content to be 60-70% of the total mass, and the kitchen waste is degraded.
Furthermore, the mass ratio of the kitchen garbage to the microbial agent is 4: 1-5: 1.
Further, the mass ratio of the microbial agent to the enzyme preparation is 20: 2-25: 2.
Further, the degradation temperature is 50-60 ℃.
The degradation reaction formula is as follows:
Figure BDA0002762995280000031
compared with the prior art, the invention has the following technical advantages:
1) according to the technical scheme, the degradation time is obviously shortened and the degradation rate is improved by compounding a plurality of specific microorganisms and a plurality of enzymes, wherein amylase, protease, cellulase and lipase decompose macromolecules of kitchen garbage into micromolecules, and the degradation time can be shortened by degrading micromolecule substances by using a microbial preparation.
2) The microbial preparation and the immobilized enzyme in the technical scheme can degrade kitchen garbage in batches continuously, so that the cost is reduced, and the utilization rate of the microbial preparation and the enzyme is improved.
Drawings
FIG. 1 is a graph showing the degradation effect of example 1 of the present invention;
FIG. 2 is a graph showing the degradation effect of example 2 of the present invention;
FIG. 3 is a graph showing the degradation effect of example 4 of the present invention.
Detailed Description
The present invention will be further explained by referring to the following embodiments, wherein the mixed microbial thallus preparation is prepared by using cells obtained by fermenting mixed strains, and the mixed microbial thallus preparation is compounded and combined with an enzyme preparation to degrade kitchen waste. The examples are set forth with the understanding that they are intended to be illustrative of the invention only and do not limit the scope of the invention.
The protease, amylase, cellulase and lipase preparations of the invention are produced by Shandong Kete enzyme preparation Co.
The combination of the microbial preparation and the immobilized enzyme provided by the invention can improve the degradation rate, increase the degradation batch and reduce the economic cost. The amylase and the cellulase decompose food into micromolecular saccharides, the protease decomposes food into micromolecular amino acids, the lipase decomposes oil food into fatty acid, and the micromolecular can be quickly degraded by microorganisms to be beneficial to growth, so that the degradation rate is improved. The immobilized enzyme can be recycled, so that the degradation batch is increased, and the cost can be reduced.
The preparation method of the microbial mixed thallus preparation comprises the following steps:
preparation of a microbial mixed thallus preparation: bacillus licheniformis CGMCC1.813, Bacillus subtilis CGMCC1.14985 and Bacillus subtilis CGMCC1.7417 are added according to the proportion of 1: 1: the ratio of 1 is inoculated in a seed liquid nutrient broth culture medium and mixed and cultured at 37 ℃ overnight. After 12h to 14h, evenly mixing and inoculating the mixture into a nutrient broth culture medium of fermentation liquor, and culturing for 15h to 18h at 37 ℃. Pouring the fermentation liquor into a container, adding wood chips into the container at a ratio of 4: 1-6: 1, mixing, and air-drying.
The preparation method of the immobilized enzyme comprises the following steps:
in the following examples, the mass ratio of amylase, cellulase, protease and lipase is as follows: 1: 1: 1: 1, proportioning.
Immobilization of the enzyme: the enzyme was resuspended in 0.8% normal saline in a ratio of 1: 5 (m: V), preparing enzyme solution, mixing the wood chips and the enzyme solution according to a certain proportion, and placing the mixture on a constant temperature shaking bed with 180rpm for shaking and adsorption for 2-3 hours. Adding glutaraldehyde with the volume fraction of about 1-3%, oscillating for 30min on an oscillator, and standing and crosslinking for 3h at 4 ℃.
The method for degrading the kitchen garbage comprises the following steps:
pouring kitchen waste into a degradation reactor, adding wood dust, wherein the adding amount of the wood dust is 50% -55% of the weight of the waste, mixing a microbial mixed thallus preparation with free enzyme or immobilized enzyme according to the ratio of 50-60: 1, the microbial mixed thallus preparation is added into a degradation reactor, the adding amount of the microbial mixed thallus preparation is 20-25% of the weight of the garbage, the garbage is degraded under the condition of 50-60 ℃, samples are taken for many times, and data are recorded. After the kitchen waste is degraded, a new batch of kitchen waste can be added for continuous degradation. Wherein, the degradation rate of the kitchen waste is (total feeding amount before treatment-residual amount after treatment)/total swill input amount) x 100%.
Example 1
In the preparation method of the mixed microbial thallus preparation, the mass ratio of the components of various strains is bacillus subtilis CGMCC 1.14985: bacillus subtilis CGMCC 1.7417: bacillus licheniformis CGMCC1.813 ═ 1: 1: 1, inoculating the mixture into a nutrient broth seed culture medium, culturing at 37 ℃ and 250rpm for 12h, then inoculating the mixture into a nutrient broth fermentation culture medium according to the inoculation amount of 1%, and culturing at 37 ℃ and 250rpm for 18 h. Pouring the fermentation liquor into a container, and adding wood chips into the container according to the proportion of 5:1 (V: m), mixing and air-drying.
The enzyme immobilization process comprises the following steps of resuspending enzyme powder in 0.8% of physiological saline according to the mass ratio of 1: 50 (m: V), preparing enzyme solution, and mixing the wood chips and the enzyme solution according to the proportion of 1: 4 (m: V), placing on a constant temperature shaking table of 180rpm, and shaking for 3 h. Glutaraldehyde was added to a final concentration of about 2%, and after shaking for 30min on a shaker, it was allowed to stand at 4 ℃ for crosslinking for 3 hours.
The degradation process is that 40g of kitchen garbage is poured into a degradation reactor, 20g of sawdust and 8g of mixed microbial thallus preparation are added, and degradation is carried out at 50 ℃. And when the first batch of garbage is completely degraded, continuously adding a second batch of garbage of 40.01g, and degrading for 24 hours. The garbage is degraded into two batches, the total degradation is 43.88g, and the degradation rate reaches 54.85%, see figure 1.
Example 2
Pouring 40g of kitchen garbage into a degradation reactor, adding wood chips, wherein the adding amount of the wood chips is 55% of the weight of the garbage, mixing a microbial preparation obtained by fermenting for 30 hours and free enzyme according to the proportion of 12:1, adding the microbial preparation-enzyme preparation compound into the degradation reactor, wherein the adding amount of the microbial preparation-enzyme preparation compound is 25% of the weight of the garbage, degrading at 50 ℃, and continuously adding a second batch of 40.03g of garbage when the first batch of garbage is completely degraded, and degrading for 24 hours. The garbage is degraded into two batches, 77.67g in total, and the degradation rate reaches 97.05 percent, see figure 2.
Example 3
Similar to the operation of example 4, except that the degradation temperature was changed to 60 ℃, after 24 hours of degradation, the garbage was degraded in two batches, total 70.5g, and the degradation rate reached 87.52%.
Example 4
Pouring 40g of kitchen garbage into a degradation reactor, adding sawdust, wherein the adding amount of the sawdust is 53% of the weight of the garbage, mixing a microorganism mixed thallus preparation obtained by fermenting for 18 hours and an immobilized enzyme according to the proportion of 12:1, and adding the microorganism-enzyme preparation compound into the degradation reactor, wherein the adding amount of the microorganism-enzyme preparation compound is 25% of the weight of the garbage. And (3) degrading for 24 hours at the temperature of 50 ℃, and continuing to add the second batch (40g), the third batch (40.01g) and the fourth batch (40.01g) of garbage for degradation when the garbage of one batch is completely degraded. The garbage is degraded into four batches, 151.58g in total, and the degradation rate reaches 94.73%, see figure 3.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.

Claims (10)

1. A microbial agent-enzyme preparation compound for degrading kitchen garbage is characterized by comprising a microbial agent and an enzyme preparation, wherein the microbial agent comprises mixed microorganisms of bacillus and wood chips, and the enzyme preparation comprises amylase, cellulase, protease and lipase.
2. The kitchen waste degrading microbial agent-enzyme preparation compound as claimed in claim 1, wherein the mass ratio of the microbial agent to the enzyme preparation is 20: 2-25: 2.
3. The microbial agent-enzyme preparation compound for degrading kitchen waste according to claim 1, wherein the mass ratio of amylase to cellulase to protease to lipase is as follows: 0.8-1.1: 0.8-1.1: 0.8-1.1: 0.8 to 1.1.
4. The kitchen waste degrading microbial agent-enzyme preparation compound as claimed in claim 1, wherein the microbial agent comprises mixed microorganisms of bacillus and wood chips in a mass ratio of 4-6: 1.
5. The kitchen waste degrading microbial agent-enzyme preparation compound as claimed in claim 1, wherein the mixed microorganism of the bacillus genus comprises bacillus licheniformis and bacillus subtilis in a mass ratio of 1: 2.
6. A method for preparing the microbial inoculant-enzyme preparation compound of claim 1, which comprises the following steps: and (3) fermenting and culturing the composite strain in a culture medium for 24-36 hours at the fermentation temperature of 37 ℃, mixing the composite strain with wood chips, air-drying to obtain a microbial agent, and adding an enzyme preparation to obtain the microbial agent-enzyme compound.
7. The method for preparing a microbial inoculant-enzyme preparation combination according to claim 6, wherein the culture medium is a nutrient broth culture medium.
8. The method for preparing a microbial agent-enzyme preparation compound according to claim 6, wherein the enzyme preparation is free enzyme or immobilized enzyme.
9. The application of the microbial agent-enzyme preparation compound in degrading kitchen garbage according to claim 1, which comprises the following steps: pouring the kitchen waste into a degradation container, adding a microbial agent and enzyme complex, adding water to ensure that the water content is between 60 and 70 percent of the total mass, and beginning to degrade the kitchen waste at the degradation temperature of 50 to 60 ℃.
10. The application of the microbial agent-enzyme preparation compound in claim 9 in degrading kitchen waste is characterized in that the mass ratio of the kitchen waste to the microbial agent is 4: 1-5: 1;
the mass ratio of the microbial agent to the enzyme preparation is 20: 2-25: 2.
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Application publication date: 20210312