CN113755361B - Preparation and application of microbial agent for continuous and efficient conversion of kitchen waste - Google Patents

Preparation and application of microbial agent for continuous and efficient conversion of kitchen waste Download PDF

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CN113755361B
CN113755361B CN202110666611.XA CN202110666611A CN113755361B CN 113755361 B CN113755361 B CN 113755361B CN 202110666611 A CN202110666611 A CN 202110666611A CN 113755361 B CN113755361 B CN 113755361B
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lactobacillus
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CN113755361A (en
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王羽贤
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Zhongyi Fangte Beijing Environmental Protection Technology Co ltd
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    • C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
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Abstract

The invention discloses preparation and application of a microbial agent for continuous and efficient conversion of kitchen waste. The microbial inoculum comprises Bacillus flexus zyftem11, and the registration number of the Bacillus flexus zyftem11 in the China general microbiological culture Collection center is CGMCC No.22217. The microbial inoculum has simple production flow, low using and operating difficulty, can realize continuous 1-month degradation of the kitchen waste after one-time addition, and has huge application prospect. The normal-temperature treatment of the kitchen waste can be realized by using the microbial inoculum, the high temperature is not needed, and the energy consumption is reduced.

Description

Preparation and application of microbial agent for continuous and efficient conversion of kitchen waste
Technical Field
The invention relates to the technical field of biology, in particular to preparation and application of a microbial agent for efficiently converting kitchen waste in a kitchen waste integrated treatment device.
Background
The kitchen waste mainly refers to kitchen waste and dining table residual waste generated in daily life, and the data show that the annual generation amount of the kitchen waste in China city is about 6 multiplied by 10 7 t, in some first-line cities with dense population, the daily output of the kitchen waste reaches or even exceeds 1000t. The kitchen waste accounts for more than 40% of the total amount of the municipal solid waste. Because of the characteristics of high water content, high oil content, high salt content, easy rancidity and degradation, bad smell and pathogenic bacteria, etc., the environment pollution is caused, and the human health is threatened. At present, the treatment technologies of the kitchen waste which are applied more in China include methods of burning and burying, forage formation, fertilizer formation, anaerobic digestion and the like. The secondary pollution problem exists in burning and landfill; the feed technology has been put out of documents to prohibit the feed application of the kitchen waste due to the existence of unpredictable unsafe factors such as homology pollution and the like; organic fertilizer products formed by high-temperature aerobic fermentation, aerobic composting and the like are applied and stored in large quantitiesThe salinization problem of soil; anaerobic fermentation is a mainstream process for treating kitchen waste in China, but has the problems of high equipment requirement, easiness in acidification, low gas production rate, difficulty in treating anaerobic digestion residues and the like.
The current technologies for microbial treatment of kitchen waste mainly focus on the following three major categories:
1) Microbial inoculum is obtained by microbial fermentation and is directly applied to kitchen waste treatment. One type of research or patent focuses on the treatment effect of the microbial inoculum and the analysis of the growth condition and the enzyme production condition of microorganisms in the treatment process; the second category concerns whether the degradation products can be used as organic fertilizers; however, the elimination of odor in the kitchen waste treatment process is not considered in the former two types, and some treatments need to be carried out at a high temperature of more than 50 ℃, so that the energy consumption is high; the third type monitors the odor removal by microbial inoculum treatment, but the treatment effect is poor. And when the kitchen waste is treated by the actual microbial agent, the microbial agent needs to be added for many times in some researches. The microbial inoculum is fixed on a plurality of carriers by an adsorbent or an immobilization technology, thereby improving the treatment effect of the microbial inoculum. One of the methods adopts various adsorbents to adsorb microorganisms, and patents report that continuous treatment of kitchen waste can be realized by using the adsorbents, but the adsorbents are bacillus and do not solve the problem of odor control in the degradation process; the second type is the immobilization of microorganisms using immobilization techniques. However, the final purpose of the two technologies is to improve the garbage degradation rate, and no consideration is given to the control of odor in the degradation process.
2) The kitchen waste is treated by compounding microorganisms and enzymes. However, in order to improve the treatment efficiency, the treatment temperature is over 50 ℃, or the kitchen waste is sterilized at high temperature in advance, and then the subsequent operation is carried out after anaerobic fermentation. No mention is made about odor control, continuous operation and subsequent treatment of the degradation products in the degradation process.
Although the currently disclosed patents all use different types of microorganisms, enzymes and combinations thereof, the simplicity, continuity and stability of the operation are not considered, and the currently disclosed patents basically include various bacteria or fungi except lactobacillus, the requirements on the conditions of the degradation process are complex, and the acceptability of smell in the degradation process and the convenience of subsequent treatment are problematic, so that improvements in the aspects of optimizing the types of microorganisms, simplifying the culture process, simplifying the operation difficulty and realizing the continuity of the operation are necessary.
Disclosure of Invention
The invention aims to solve the defects of the prior art, and provides a microbial agent which is added at one time, kitchen waste can be added in batches for multiple times according to actual conditions, and the preparation and the application of the high-efficiency microbial agent can realize continuous degradation for at least 1 month. The specific technical scheme is as follows:
the invention provides a microbial agent for continuously treating kitchen waste, which comprises bacillus flexus zyftem11, wherein the registration number of the bacillus flexus zyftem11 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No.22217.
Further, the microbial inoculum comprises a strain compound consisting of lactobacillus plantarum, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus fermentum, bacillus lysinate, lysine bacillus fusiformis, bacillus curvatus, bacillus natto and saccharomyces cerevisiae.
Further, the colony ratio of lactobacillus plantarum, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus fermentum, bacillus lysinate, lysine bacillus fusiformis, bacillus curvatus, bacillus natto and saccharomyces cerevisiae in the strain compound is 1.
Further, the microbial inoculum comprises protease, cellulase and lipase. The microbial inoculum comprises fermentation liquor of a strain compound, wherein the fermentation liquor contains protease, cellulase and lipase.
Further, the pH of the microbial inoculum is 4.5 to 5.5.
The above mentioned bacterial complexes are also intended to be within the scope of the present invention.
The invention also provides a Bacillus flexus zyftem11, which has the registration number of CGMCC No.22217 in the China general microbiological culture Collection center.
The application of the microbial agent, the strain compound and the campylobacter in degrading the kitchen waste is also within the protection scope of the invention.
The invention also provides a method for degrading the kitchen waste, and the method for degrading the kitchen waste by using the microbial inoculum comprises the following steps:
1) Adding a filler into the continuous kitchen waste treatment equipment;
2) Adding a microbial inoculum according to the proportion of adding 1000mL of microbial inoculum to each 1000g of kitchen waste to be treated, and mixing the microbial inoculum with a filler for 30 minutes;
3) Adding the kitchen waste, operating the equipment, starting to treat the kitchen waste, and after 24-48 hours, treating the kitchen waste to ensure that the weight loss rate is more than 95 percent.
4) After the equipment runs for 24 hours, the kitchen waste can be added according to the time interval of three actual meals or every 24 hours for continuous treatment, and residues in the equipment can be cleaned after the continuous treatment for 1 month.
The innovation of the invention is that:
1. the invention adopts static culture when producing the microbial inoculum, and the culture process is simple and easy to operate.
2. The degrading microbial inoculum mainly comprises lactic acid bacteria and fermentation acid-producing strains, and the pH value in the kitchen waste treatment process is weak and acidic, so that the growth of putrefying bacteria can be inhibited, and the blockage of a sewer pipeline can be prevented.
3. The lactobacillus acidophilus, the bacillus licheniformis and the bacillus natto in the degrading microbial agent can generate lipase, the lactobacillus fermentum can generate cellulase, and the bacillus licheniformis and the bacillus natto can generate protease, amylase and the like, so that the degrading microbial agent has good degrading capability on grease, cellulose, protein and the like in kitchen waste.
4. The obtained microbial inoculum does not need treatment such as carrier, adsorbent, drying and the like, and can be directly applied to relevant equipment.
5. The microbial inoculum has simple production flow, low using and operating difficulty, can realize continuous 1-month degradation of the kitchen waste after one-time addition, and has huge application prospect. The normal-temperature treatment of the kitchen waste can be realized by using the microbial inoculum, the high temperature is not needed, and the energy consumption is reduced.
6. The lactobacillus plantarum, lactobacillus paracasei, lysine bacillus fusiformis and saccharomyces cerevisiae in the degradation microbial inoculum can effectively reduce the peculiar smell in the treatment process, and no odor is generated in the whole treatment period.
7. The composite microbial inoculum prepared by the invention contains amylase, protease, cellulase and lipase generated by microbial fermentation besides live bacteria, and is more beneficial to the rapid decomposition of organic materials.
Deposit description
The strain name: bacillus flexus
Latin name: bacillus flexus
The strain number is as follows: zyftem11
The preservation organization: china general microbiological culture Collection center
The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)
Address: xilu No. 1 Hospital No. 3 of Beijing market facing Yang district
The preservation date is as follows: 2021, 05 month and 06 days
Registration number of the preservation center: CGMCC No.22271
Drawings
FIG. 1 shows the liquid inoculum cultured in a triangular flask.
FIG. 2 shows the compounded bacterial preparation after 50 times concentration.
FIG. 3 is a graph showing the effect of Chinese cabbage before and after 24 hours treatment (total 48 hours treatment).
FIG. 4 shows the kitchen waste after the microbial inoculum is continuously treated for 7 days.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples, unless otherwise specified, were carried out in a conventional manner according to the techniques or conditions described in the literature in this field or according to the product instructions. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The strains in the following examples: the lactobacillus plantarum is purchased from China center for culture Collection of industrial microorganisms (CICC 20242); the lactobacillus paracasei is purchased from China general microbiological culture Collection center (CGMCC 1.12731); the lactobacillus acidophilus is purchased from China center for culture Collection of industrial microorganisms (CICC 20244); the lactobacillus fermentum is purchased from China center for culture collection and management of industrial microorganisms (CICC 21828); the lysine bacillus is purchased from China Industrial microorganism strain preservation management center (CICC 10478); the lysinibacillus fusiformis is purchased from China industrial microorganism culture collection management center (CICC 22657); the saccharomyces cerevisiae is purchased from China center for Industrial culture Collection of microorganisms (CICC 1015); the bacillus natto is purchased from Nankai university microorganism resource Collection (NKCCMR NK 2. Original nat); the campylobacter is campylobacter zyftem11 which is a strain autonomously separated by the inventor, and the registration number of the campylobacter is CGMCC No.22217 in China general microbiological culture Collection center.
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto.
Example 1: the preparation method of the microbial agent comprises the following steps:
1) Expanded culture of kitchen waste degrading strain
The culture medium is sterilized at 115 ℃ for 30min for later use, and then storedStrains at-80 ℃ are respectively inoculated into a culture medium, wherein lactobacillus plantarum is inoculated into an MRS culture medium, lactobacillus paracasei is inoculated into an Elliker culture medium, lactobacillus acidophilus is inoculated into a BCP culture medium, lactobacillus fermentum is inoculated into an HP culture medium, bacillus lysinate, bacillus fusiformis, bacillus flexus zyftem11 and bacillus natto are respectively inoculated into an EP culture medium, and saccharomyces cerevisiae is inoculated into an YPD culture medium, and the inoculation proportion is 1%. The Lactobacillus and the yeast are statically cultured for 24h at 37 ℃ and 28 ℃ respectively. The bacillus was shake-cultured at 28 ℃ and 140rpm for 24h. The concentration of each strain after culture is 10 8 Per liter (as shown in figure 1). The prepared lactobacillus plantarum, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus fermentum, bacillus lysinate, lysine bacillus fusiformis, bacillus flexus zyftem11, bacillus natto and saccharomyces cerevisiae culture solution are uniformly mixed according to a volume ratio of 1 9 And (2) per liter. The pH value of the microbial inoculum is 5.5, and the amylase activity in the microbial inoculum is as follows: 38U/mL; and (3) cellulase activity: 25U/mL; protease activity: 51U/mL; lipase activity: 42U/mL (as shown in FIG. 2).
The media involved are as follows:
the method comprises the following steps of: 10g of peptone, 10g of beef extract powder, 5g of yeast extract powder, 2g of diammonium hydrogen citrate, 20g of glucose monohydrate, 5g of sodium acetate trihydrate, 2g of dipotassium hydrogen phosphate trihydrate, 0.58g of magnesium sulfate heptahydrate, 0.25g of manganese sulfate monohydrate, 801mL of Tween-801, and deionized water to a constant volume of 1L, wherein the pH value is 6.0-6.0.
And an Elliker culture medium: 20g of tryptone, 5g of sucrose, 5g of yeast extract powder, 4g of sodium chloride, 2.5g of gelatin, 1.5g of sodium acetate trihydrate, 5g of glucose monohydrate, 0.5g of ascorbic acid, 5g of lactose and 1L of deionized water, wherein the pH value is 6.8.
Fourthly, BCP culture medium: 25g of yeast powder, 5g of tryptone, 5g of glucose monohydrate and deionized water to a constant volume of 1L, wherein the pH value is 7.0.
The HP culture medium: 20g of plant peptone, 5g of ammonium citrate, 6g of yeast powder, 0.04g of ferrous sulfate, 10g of beef extract, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate monohydrate, 10g of glucose monohydrate, and 801ml of tween-801, deionized water is added to the volume of 1L, and the pH value is 6.6.
Sixthly, spore bacteria (EP) culture medium: peptone 5.0g, beef extract 2.5g, mgSO 4 0.5g、K 2 HPO 4 10.0g、(NH 4 ) 2 SO 4 2.0g, naCl 2.5g, deionized water to volume of 1L, and pH7.2-7.4.
Yeast-quieter YPD medium: 20g of peptone, 10g of yeast extract, 20g of monohydrate glucose and deionized water are added to the volume of 1L, and the pH is natural.
Example 2: degradation effect test of waste vegetable leaves
1000g of waste Chinese cabbages collected in a kitchen are cut into random small blocks, and 500mL of the microbial agent prepared in the example 1 is put into special kitchen waste continuous degradation equipment; degrading continuously at room temperature for 48h, wherein the residue content in the container is 0-17.5 g, and the degradation process has slight lactic acid fermentation taste and no odor after 24h (as shown in figure 3). The degradation rate of the microbial agent for degrading the kitchen waste is calculated according to the following formula:
degradation rate = (1-M2/M1). Times.100%
Wherein M1 is the amount of the kitchen waste added before degradation, and M2 is the amount of residues in the degraded container.
According to the formula, the degradation rate in the embodiment is 98.3% -100%, and the pH value of the effluent is 6.41.
Example 3: continuous degradation effect test of kitchen waste
400mL of the microbial agent prepared in example 1 is placed in a special kitchen waste continuous degradation device and is left to stand for half an hour, 410.9 g of kitchen waste (mainly containing high starch and a small amount of meat) is added, after 24 hours of continuous degradation at room temperature, 477.6 g of kitchen waste (mainly containing high starch and a small amount of pericarp) is placed in the kitchen waste for the second time, 195.1 g of kitchen waste (mainly containing flour and a small amount of fructose) is placed in the kitchen waste for the third time after 24 hours of continuous degradation at room temperature, 409.6 g of kitchen waste (mainly containing flour) is placed in the kitchen waste for the fourth time after 24 hours of continuous degradation at room temperature, 4324 g zxft 4324 g (containing flour, meat and vegetables) is placed in the kitchen waste for the fifth time after 3 days of continuous degradation at room temperature, 423.2 g of kitchen waste (containing starch) is placed in the kitchen waste for the sixth time after 2 days of continuous degradation at room temperature, 3732 g of kitchen waste (containing flour, 3732 g of vegetables) is placed in the kitchen waste for the seventh time after 1 day of continuous degradation at room temperature, 3732 g of kitchen waste, and the kitchen waste for eight days of continuous degradation at room temperature. The total amount of kitchen garbage put into the kitchen garbage is 5000 g or so for eight times, 215.2 g of plant cellulose residues are left in the equipment after degradation, no odor is generated in the whole degradation process, and the kitchen garbage has a slight lactic acid fermentation taste (the taste of slightly pickled pickles) after the degradation is started for 24 hours. According to the formula in example 2, the degradation rate in this example was 95.7%, and the pH of the effluent was 6.40.
Example 4: continuous degradation effect test of kitchen waste
Placing 1000mL of the microbial agent prepared in the example 1 into a special kitchen waste continuous degradation device, standing for half an hour, adding 1000g of cabbage leaves, continuously degrading for 24 hours at room temperature, then adding 500g of fruit peels and vegetable leaves for the second time, continuously degrading for 24 hours at room temperature, then adding 500g of Chinese cabbage for the third time, 794.35 g of kitchen waste (starch is the main component and a small amount of meat), continuously degrading for 48 hours at room temperature, then adding 507.8 g of kitchen waste (starch is the main component and a small amount of meat), 250g of vegetable leaves, continuously degrading for 24 hours at room temperature, then adding 578 g of kitchen waste (starch and meat), 250g of vegetable leaves, and continuously degrading for 48 hours. 3801 g of kitchen waste is put into the kitchen waste for five times, 201.6 g of plant cellulose residues are left after degradation, no odor is generated in the degradation process, and a slight lactic acid fermentation taste exists after 24 hours of degradation. According to the formula in example 2, the degradation rate in this example is 94.7%, and the pH of the effluent is 6.41 (as shown in FIG. 4).
Example 5: evaluation of continuous degradation effect of kitchen waste for 1 month
This example is a family effect lab scale of one family (parents are on duty, children are on school, elderly are at home, breakfast and dinner are taken at one family, and only 1 elderly is at home for Chinese meal). Putting 1000mL of the microbial agent prepared in the example 1 into special kitchen waste continuous degradation equipment, standing for half an hour, adding 1000g of cabbage leaves, and continuously degrading for 24 hours at room temperature; then adding the kitchen waste, wherein the time for adding the kitchen waste is 7% in the morning each day: about 30, at noon 12. The adding duration is from 18 days 6 months to 20 days 7 months in 2020 for 33 days. The kitchen waste types comprise leftovers, vegetable leaves, leftovers, fruit peels, meat and grease. 181.2 g of residual plant cellulose residue after degradation. Opening the opening of the equipment after adding the kitchen waste for 24 hours, wherein the opening has slight sour taste, and no obvious smell or stink is generated in the whole degradation process. The amount of garbage added every day is estimated to be about 300g, and the degradation rate is estimated to be about 98%.
Example 6: kitchen waste continuous degradation effect test without adding microbial inoculum
In a special kitchen waste continuous degradation device, bacterial agents are not added, 1000g of Chinese cabbage leaves are directly added, 500g of fruit peels and vegetable leaves are thrown in for the second time after continuous natural degradation is carried out for 24 hours at room temperature, 500g of Chinese cabbage is thrown in for the third time after continuous natural degradation is carried out for 24 hours at room temperature, 794.35 g of kitchen waste (starch is the main component and a small amount of meat) is thrown in for the third time, 507.8 g of kitchen waste (starch is the main component and a small amount of meat) is thrown in for the fourth time after continuous natural degradation is carried out for 48 hours at room temperature, 250g of vegetable leaves are thrown in for the fifth time after continuous natural degradation is carried out for 24 hours at room temperature, 578 g of kitchen waste (starch and meat), 250g of vegetable leaves are thrown in for the fifth time, and the continuous natural degradation is carried out for 48 hours. 3801 g of kitchen waste and 2050 g of residues in equipment are put into the kitchen waste for five times, and the natural degradation rate is 46.1%. After natural degradation for 24h, a malodorous smell began to appear, and a strong malodorous smell began to be generated on day 2.
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is made possible within the scope of the claims attached below.

Claims (5)

1. The microbial agent for continuously treating the kitchen waste is characterized by comprising bacillus flexus zyftem11, wherein the registration number of the bacillus flexus zyftem11 in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms is CGMCC No.22217; the microbial inoculum comprises a strain compound consisting of lactobacillus plantarum, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus fermentum, bacillus lysinate, lysine bacillus fusiformis, bacillus flexus zyftem11, bacillus natto and saccharomyces cerevisiae; the colony ratios of lactobacillus plantarum, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus fermentum, bacillus lysinate, lysine bacillus fusiformis, bacillus flexus zyftem11, bacillus natto and saccharomyces cerevisiae in the strain compound are 1; the microbial inoculum comprises protease, cellulase and lipase; the pH of the microbial inoculum is 4.5 to 5.5; wherein the lactobacillus plantarum has a preservation number of CICC 20242 in China center for culture collection and management of industrial microorganisms; the preservation number of the lactobacillus paracasei in the China general microbiological culture collection center is CGMCC 1.12731; the lactobacillus acidophilus is deposited in China industrial microorganism culture collection management center with the deposit number of CICC 20244; the lactobacillus fermentum has a deposit number of CICC 21828 in China center for the preservation and management of industrial microorganism strains; the deposit number of the lysine bacillus in China industrial microorganism strain preservation management center is CICC 10478; the deposit number of the lysinibacillus fusiformis in China industrial microorganism strain preservation management center is CICC 22657; the preservation number of the saccharomyces cerevisiae in the China industrial microorganism strain preservation management center is CICC 1015; the deposit number of the bacillus natto in the south China university microorganism resource preservation center is NKCCMR NK 2.
2. The Bacillus flexus strain of claim 1, zyftem11 having accession number CGMCC No.22217, which is registered at the China Committee for culture Collection of microorganisms.
3. Use of the microbial agent of claim 1 or the bacillus curvatus of claim 2 for degrading kitchen waste.
4. A method for degrading kitchen waste, which is characterized by using the microbial inoculum described in claim 1.
5. The method of claim 4, comprising the steps of:
1) Adding a filler into the kitchen waste continuous treatment equipment;
2) Adding a bacterial agent according to the proportion of adding 1000mL bacterial agent to each 1000g kitchen waste to be treated, and mixing the bacterial agent with the filler for 30 minutes;
3) Adding kitchen waste, operating equipment, and beginning to treat the kitchen waste, wherein the weight loss rate of the kitchen waste after treatment is more than 95% after 24-48 hours;
4) After the equipment runs for 24 hours, the kitchen waste is added according to the time interval of three actual meals or every 24 hours for continuous treatment, and residues in the equipment can be cleaned after the continuous treatment for 1 month.
CN202110666611.XA 2021-06-16 2021-06-16 Preparation and application of microbial agent for continuous and efficient conversion of kitchen waste Active CN113755361B (en)

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