CN112760274B - Organic solid waste high-temperature aerobic composting strain and application thereof - Google Patents

Organic solid waste high-temperature aerobic composting strain and application thereof Download PDF

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CN112760274B
CN112760274B CN202110370552.1A CN202110370552A CN112760274B CN 112760274 B CN112760274 B CN 112760274B CN 202110370552 A CN202110370552 A CN 202110370552A CN 112760274 B CN112760274 B CN 112760274B
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pantoea
temperature
microbial inoculum
organic solid
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CN112760274A (en
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刘莉
吉雨霁
史吉平
王娜
张斯颖
陈晓淼
肖梦瑶
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Shanghai Advanced Research Institute of CAS
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    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • 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
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F3/00—Fertilisers from human or animal excrements, e.g. manure
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • C05F5/002—Solid waste from mechanical processing of material, e.g. seed coats, olive pits, almond shells, fruit residue, rice hulls
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F7/00—Fertilisers from waste water, sewage sludge, sea slime, ooze or similar masses
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F9/00—Fertilisers from household or town refuse
    • C05F9/04—Biological compost
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • 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
    • Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Abstract

The invention belongs to the technical field of environmental microorganisms, and particularly relates to an organic solid waste aerobic composting strain which is identified as PantoeaPantoeasp, the preservation number is CCTCC NO: M2020973. Said Pantoea genusPantoeaThe sp.BK-PP311 is high-temperature resistant, has the capability of efficiently degrading organic matters and quickly decomposing, and can be applied to the high-temperature aerobic composting treatment process of organic solid wastes such as food wastes, kitchen wastes, fruit and vegetable wastes, livestock and poultry breeding manure, municipal sludge and the like, thereby realizing the quick composting and resource utilization of the organic solid wastes.

Description

Organic solid waste high-temperature aerobic composting strain and application thereof
Technical Field
The invention relates to the technical field of environmental microorganisms, in particular to an organic solid waste high-temperature aerobic composting strain and application thereof.
Background
In recent years, with the growth of population and the development of agriculture and industry, the yield of organic solid wastes has been increasing year by year. The sources of the organic solid wastes mainly comprise food wastes, kitchen wastes, fruit and vegetable wastes, livestock and poultry breeding manure, municipal sludge and the like. At present, a large amount of organic solid wastes are not fully utilized, and the resource waste is serious. Therefore, how to improve the resource utilization rate of the organic solid wastes and realize the reduction, harmless treatment and resource utilization of the organic solid wastes is urgent.
At present, the resource utilization approaches of organic solid wastes comprise energy regeneration, fertilizer conversion, feed conversion and the like. Wherein, the organic solid waste is converted into the organic fertilizer in an aerobic composting way, which is a main way for realizing the resource utilization of the waste. Aerobic composting refers to a process of stabilizing organic substances in organic solid waste under the action of aerobic microorganisms under aerobic conditions, wherein thallus metabolism generates certain heat in the composting process, and harmless treatment and stabilization can be realized without an external heating source. The traditional aerobic composting technology has the problems of short high-temperature period, long composting period, incomplete fermentation, serious nutrient loss, low material decomposition degree and the like. Compared with the traditional compost, the high-temperature compost has the following advantages: shortening the composting period, accelerating the degradation of organic matters, promoting the rapid fermentation and decomposition of the compost, improving the temperature of the compost, effectively killing pathogenic bacteria and ova, degrading residual antibiotics in the compost and the like. High-temperature bacteria are required to be added into the compost body, the high-temperature bacteria can prolong the high-temperature period, accelerate the degradation and conversion of organic matters, promote the compost maturity and shorten the composting period, so that the composting efficiency and the product quality are greatly improved. Therefore, the method has great significance for screening organic solid waste high-temperature composting strains and researching and developing high-temperature composting microbial agents and accelerating the fertilizer utilization of organic solid waste.
Disclosure of Invention
In view of the above disadvantages of the prior art, the present invention provides an organic solid waste high temperature aerobic compost strain and its application, which are used to solve the problems in the prior art.
To achieve the above and other related objects, the present invention provides PantoeaPantoeasp, BK-PP311, with the preservation number of CCTCC NO: m2020973.
The invention also provides a liquid microbial inoculum, which comprises the pantoeaBelong toPantoea sp.BK-PP311, a strain of the genus Pantoea in the liquid inoculumPantoea sp. The concentration of BK-PP311 is at least 1 x 108 cfu/mL。
The invention also provides a preparation method of the liquid microbial inoculum, which comprises the following steps: subjecting said Pantoea toPantoea spInoculating the pure strain of BK-PP311 in a liquid culture medium for culturing to obtain the liquid microbial inoculum after the culture is finished.
The invention also provides a solid microbial inoculum, which comprises the PantoeaPantoeasp, BK-PP311 and a carrier.
The invention also provides a preparation method of the solid microbial inoculum, which comprises the following steps: and mixing the carrier and the liquid microbial inoculum according to the mass ratio of 1: 1-1: 10, and drying to obtain the solid microbial inoculum.
The present invention also provides the Pantoea genusPantoea sp.The BK-PP311 is applied to organic solid waste aerobic composting.
The invention also provides a method for fermenting the organic solid waste aerobic compost, which comprises the following steps: and mixing the solid microbial inoculum with organic solid waste raw materials, and then carrying out aerobic composting fermentation.
As mentioned above, the organic solid waste high-temperature aerobic composting strain and the application thereof have the following beneficial effects:
(1) pantoea genus of the present inventionPantoeasp, BK-PP311, which has high temperature resistance and can carry out rapid aerobic composting fermentation on organic solid waste at high temperature (50-75 ℃);
(2) pantoea genusPantoeasp, BK-PP311 also has the capability of high yield of protease and amylase under the high temperature condition, and can efficiently degrade organic matters such as protein, starch and the like in organic solid waste. Pantoea of the inventionPantoeaThe addition of sp, BK-PP311 can lead the decomposition and decomposition of organic matters to be more thorough in the aerobic composting fermentation process, and the prepared organic fertilizer has higher content of nutrient components.
By combining the above two characteristics, PantoeaPantoeaThe sp, BK-PP311 is applied to high-temperature aerobic composting of organic solid waste, and can realize the harmlessness, stabilization and resource of the organic solid wasteAnd (4) transforming.
Drawings
FIG. 1 shows a phylogenetic tree of the strain BK-PP311 of the invention;
FIG. 2 shows the change of the water content of the food waste and wheat bran high-temperature aerobic composting material by the strain BK-PP 311;
FIG. 3 shows the temperature change of the fermentation heap of the food waste and wheat bran high-temperature aerobic composting caused by the strain BK-PP 311;
FIG. 4 shows the change of the water content of the high-temperature aerobic composting materials of fruit and vegetable garbage and pig manure by the strain BK-PP 311;
FIG. 5 shows the temperature change of the high-temperature aerobic composting fermentation heap of the fruit and vegetable garbage and pig manure by the strain BK-PP 311.
Detailed Description
The invention obtains the organic solid waste high-temperature compost strain through preliminary screening of a separation culture medium, secondary screening of an enzyme production culture medium and high-temperature aerobic composting experimental verification.
The invention provides a high-temperature aerobic composting strain for organic solid wastes, which is characterized in that the taxonomic status of the strain is identified through comprehensive analysis of data such as colony and cell morphology, physiological and biochemical characteristics, 16S rDNA gene sequence determination and the like: pantoea genusPantoeasp. storing the strain in China Center for Type Culture Collection (CCTCC) with the strain name ofPantoeasp.BK-PP311, preservation date is 2020.12.28, and preservation number is CCTCC NO: m2020973, the preservation address is Wuhan university in China.
Said Pantoea genusPantoeasp.BK-PP311 has a gene sequence shown in SEQ ID NO. 1.
Said Pantoea genusPantoeasp.BK-PP311 is gram-negative bacteria and rod-shaped, and the colony morphology is approximately circular, milky white, irregular edge, rough surface and wrinkle.
Said Pantoea genusPantoeasp.BK-PP311 is resistant to high temperatures. Specifically, the Pantoea genusPantoeasp.The maximum growth temperature of BK-PP311 was 75 ℃. Above 75 ℃ the strain does not grow.
The organic solid waste comprises one or more of food waste, kitchen waste, fruit and vegetable waste, livestock and poultry breeding manure and municipal sludge.
In one embodiment, the Pantoea genus can bePantoeasp.BK-PP311 is prepared into liquid microbial inoculum or solid microbial inoculum for aerobic composting.
Using said Pantoea genusPantoeasp.When BK-PP311 is used for aerobic composting of organic solid wastes, the temperature of the compost can reach 75 ℃. Using said Pantoea genusPantoeasp.When BK-PP311 is subjected to aerobic composting, the high-temperature maintenance period is prolonged by more than 3 days compared with that of a control group.
The high-temperature maintenance period refers to the days when the temperature of the stack reaches 55 ℃ or above.
The stack temperature and the high-temperature maintenance period are obtained through the following steps:
1) mixing organic solid wastes with PantoeaPantoeasp. BK-PP311 solid microbial inoculum mixture, PantoeaPantoeasp. The mass of the BK-PP311 solid microbial inoculum is 0.2-0.5% of that of the organic solid waste;
2) mixing, placing in an aerobic fermentation device, and fermenting, wherein the materials are stirred for 10 min every 2 h in the fermentation process, and the ventilation rate is 30 mL/min;
3) during the fermentation process, 9: 00 the temperature at the center of the stack is measured. Control group without PantoeaPantoeasp. Except the BK-PP311 solid microbial inoculum, other operations are completely the same.
The organic solid waste is selected from one or more of food waste, fruit and vegetable waste and livestock and poultry breeding manure.
In a second aspect, the present invention provides a liquid microbial inoculum comprising a Pantoea genusPantoeasp.BK-PP311, a strain of the genus Pantoea in the liquid inoculumPantoeasp.The concentration of BK-PP311 is at least 1 x 108 cfu/mL。
The third aspect of the invention provides a preparation method of the liquid microbial inoculum, which comprises the following steps: pantoea genusPantoeasp.Inoculating the pure strain of BK-PP311 in a liquid culture medium for culture, and obtaining the liquid microbial inoculum after the culture is finished.
In one embodiment, Pantoea henryi is culturedBelong toPantoeasp.Inoculating the pure strain of BK-PP311 into a liquid culture medium for culture, then inoculating the obtained culture solution into another liquid culture medium for amplification culture, and obtaining the liquid microbial inoculum after the culture is finished.
In one embodiment, the temperature of the culture is 40-60 ℃.
In one embodiment, the stirring speed of the culture is 150 to 230 r/min.
In one embodiment, Pantoea is treatedPantoeasp.Inoculating the pure strain of BK-PP311 into a liquid culture medium, culturing for 12-16 h, and then inoculating into another liquid culture medium for amplification culture.
In one embodiment, the liquid microbial inoculum of the strain can be obtained after the expansion culture for 16-20 hours.
The liquid culture medium consists of: 3 g/L of peptone and 2 g/L, NaCl 3 g/L, MgCl of yeast extract2 0.125 g/L、CaCl2 0.5 g/L、FeSO4·7H20.01 g/L of O, trace element solution (Na)2MoO4 12 g/L、VOSO4·xH2O 1 g/L、MnCl2 5 g/L、ZnSO4·7H2O 0.6 g/L、CuSO4·5H2O 0.15 g/L、CoCl2·6H2O 8 g/L、NiCl2·6H2O0.2 g/L) 100 mu L/L, and the pH value is adjusted to 7-7.5.
In a fourth aspect, the present invention provides a solid microbial agent comprising said PantoeaPantoeasp.BK-PP311 and a carrier.
The carrier is a solid phase carrier. The carrier is selected from one or more of starch, cyclodextrin, xanthan gum, calcium carbonate, rice hull, wheat bran and the like.
The fifth aspect of the invention provides a preparation method of the solid microbial inoculum, which comprises the steps of mixing a carrier and a liquid microbial inoculum according to the mass ratio of 1: 1-1: 10, and drying to obtain the solid microbial inoculum.
The sixth aspect of the present invention provides the genus PantoeaPantoeasp. The BK-PP311 is applied to organic solid waste aerobic composting.
The organic solid waste comprises one or more of food waste, kitchen waste, fruit and vegetable waste, livestock and poultry breeding manure and municipal sludge.
Specifically, the aerobic compost is high-temperature aerobic compost.
The high-temperature aerobic composting is selected from any one of a closed type, a strip stack type and a groove type.
The seventh aspect of the invention provides a method for organic solid waste aerobic composting fermentation, which comprises the following steps: and mixing the solid microbial inoculum with organic solid waste raw materials, and then carrying out aerobic composting fermentation.
In one embodiment, the mass of the solid microbial inoculum is 0.1-1% of the mass of the organic solid waste raw material;
in one embodiment, the aerobic composting fermentation process is aerated and agitated. The ventilation stirring can ensure that the temperature of the materials is uniform and the materials are carried out under aerobic conditions.
The aerobic composting fermentation time can be specifically determined according to different materials. In one embodiment, the aerobic composting fermentation time is 10 days or more. The aerobic fermentation time is, for example, 10 to 15 days, 15 to 20 days, 20 to 25 days or longer.
The organic solid waste comprises one or more of food waste, kitchen waste, fruit and vegetable waste, livestock and poultry breeding manure and municipal sludge.
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
The following are the media and formulations used in the examples of the invention:
separating and purifying the culture medium: 3 g/L of peptone and 2 g/L, NaCl 3 g/L, MgCl of yeast extract2 0.125 g/L、CaCl2 0.5 g/L、FeSO4·7H20.01 g/L of O, trace element solution (Na)2MoO4 12 g/L、VOSO4·xH2O 1 g/L、MnCl2 5 g/L、ZnSO4·7H2O 0.6 g/L、CuSO4·5H2O 0.15 g/L、CoCl2·6H2O 8 g/L、NiCl2·6H2O0.2 g/L) 100 mu L/L, agar 20 g/L, and the pH value is adjusted to 7-7.5.
Seed culture medium: 3 g/L of peptone and 2 g/L, NaCl 3 g/L, MgCl of yeast extract2 0.125 g/L、CaCl2 0.5 g/L、FeSO4·7H20.01 g/L of O, trace element solution (Na)2MoO4 12 g/L、VOSO4·xH2O 1 g/L、MnCl2 5 g/L、ZnSO4·7H2O 0.6 g/L、CuSO4·5H2O 0.15 g/L、CoCl2·6H2O 8 g/L、NiCl2·6H2O0.2 g/L) 100 mu L/L, and the pH value is adjusted to 7-7.5.
Protease screening medium: casein 10 g/L, peptone 3 g/L, yeast extract 2 g/L, NaCl 3 g/L, M g/LgCl2 0.125 g/L、CaCl2 0.5 g/L、FeSO4·7H20.01 g/L of O, trace element solution (Na)2MoO4 12 g/L、VOSO4·xH2O 1 g/L、MnCl2 5 g/L、ZnSO4·7H2O 0.6 g/L、CuSO4·5H2O 0.15 g/L、CoCl2·6H2O 8 g/L、NiCl2·6H2O0.2 g/L) 100 mu L/L, agar 20 g/L, and the pH value is adjusted to 7-7.5.
Amylase screening culture medium: 10 g/L of soluble starch, 3 g/L of peptone and 2 g/L, NaCl 3 g/L, MgCl of yeast extract2 0.125 g/L、CaCl2 0.5 g/L、FeSO4·7H20.01 g/L of O, trace element solution (Na)2MoO4 12 g/L、VOSO4·xH2O 1 g/L、MnCl2 5 g/L、ZnSO4·7H2O 0.6 g/L、CuSO4·5H2O 0.15 g/L、CoCl26H2O 8 g/L、NiCl2 6H2O0.2 g/L) 100 mu L/L, agar 20 g/L, and the pH value is adjusted to 7-7.5.
The liquid culture medium consists of: 3 g/L of peptone and 2 g/L, NaCl 3 g/L, MgCl of yeast extract2 0.125 g/L、CaCl2 0.5 g/L、FeSO4·7H20.01 g/L of O, trace element solution (Na)2MoO4 12 g/L、VOSO4·xH2O 1 g/L、MnCl2 5 g/L、ZnSO4·7H2O 0.6 g/L、CuSO4·5H2O 0.15 g/L、CoCl2·6H2O 8 g/L、NiCl2·6H2O0.2 g/L) 100 mu L/L, and the pH value is adjusted to 7-7.5.
The prepared culture medium is sterilized by high pressure steam at 121 deg.C for 15 min.
Example 1: separation screening and performance determination of strain BK-PP311
The separation and purification process of the strain BK-PP311 of the invention comprises the following steps: collecting test samples from high-temperature composting equipment of excrement of a certain cattle farm, adding sterile water, uniformly mixing, carrying out gradient dilution, coating 200 mu L of diluent with different gradients on a separation and purification culture medium plate, culturing in a constant-temperature incubator at 60 ℃ for 24-48 h, selecting single colonies with different shapes and sizes, marking, purifying, numbering and preserving, and carrying out primary screening to obtain 26 strains.
The first secondary screening: and inoculating the 26 pure strains obtained by primary screening to a protease screening culture medium and a amylase screening culture medium by adopting a point-to-point method, and re-screening the enzyme production capacity of the strains. After culturing at 60 ℃ for 48 h, the colony diameter (D) and the transparent circle diameter (D) are measured, D/D is calculated, and strains with stronger protein and starch degradation capacities are selected.
And (3) second re-screening: and (3) inoculating 26 strains obtained by primary screening on a separation and purification culture medium, respectively placing in incubators at 60, 65, 70, 75, 80 and 85 ℃, recording the growth conditions, carrying out secondary screening at the highest growth temperature of the strains, and selecting the strains with high temperature resistance.
And combining the two re-screening results to obtain 3 strains of the high-temperature-resistant strain with stronger protein, starch and cellulose degradation capacities.
Inoculating the strain obtained by re-screening to a seed culture medium, culturing for 16 h at 50 ℃ and 180 r/min, then inoculating to a liquid culture medium according to the inoculum size of 1% of the total volume of the liquid culture medium, carrying out amplification culture, and culturing for 16 h at 50 ℃ and 180 r/min to obtain the liquid microbial inoculum of the strain. Taking 250 mL of liquid microbial inoculum, centrifuging, washing with sterile normal saline, resuspending to 5 mL, adding into a mixture of 1000 g of food waste (taken from a dining room of a high research institute, Shanghai, China academy of sciences) and wheat bran (the mass ratio of the food waste to the wheat bran is 4: 1), stirring uniformly, and carrying out high-temperature aerobic composting experimental verification, wherein the materials are stirred for 10 min every 2 h in the process, and the ventilation rate is 30 mL/min. As a result, the germination index of the seeds after the compost of the food waste and wheat bran mixed material inoculated with the strain BK-PP311 (the determination method and the calculation method are the same as DB 37/T4135-2020) is the highest and reaches 91.8 percent after 22 d.
The highest growth temperature of the strain BK-PP311 is 75 ℃, and the strain does not grow when the temperature is higher than 75 ℃; the strain BK-PP311 has stronger protein, starch and cellulose degradation capability.
Principal of the Strain BK-PP311The biological characteristics are gram negative bacteria and rod shape, the colony morphology is approximately round and milk white, the edge is irregular, the surface is rough and wrinkled. The 16S rDNA sequence of the strain BK-PP311 is shown as SEQ ID NO.1, the 16S rDNA sequence is submitted to NCBI, homology sequence comparison analysis is carried out between software and GenBank, and MEGA X software is used for constructing a phylogenetic tree of the strain (figure 1). From the above information, it was identified that the strain BK-PP311 is Pantoea (Pantoeasp.）。
Example 2: preparation of microbial inoculum of strain BK-PP311
Preparing a liquid microbial inoculum: inoculating BK-PP311 pure strain into 10 mL seed culture medium, culturing for 16 h in a constant temperature shaking table at 50 ℃ and 180 r/min, then inoculating 1% of the strain amount into the same liquid culture medium for amplification culture, and culturing for 16 h at 50 ℃ and 180 r/min to obtain the liquid microbial inoculum of the strain.
Preparing a solid microbial inoculum: the bacterial strain is prepared by mixing starch and a liquid microbial inoculum of the bacterial strain according to the mass ratio of 1:5 and spray drying.
Example 3: high-temperature aerobic composting effect of strain BK-PP311 on food waste and wheat bran
Mixing food waste 8 kg and testa Tritici 2 kg, and adding Pantoea at a mass ratio of 0.5% to obtain a mixture of 10 kgPantoeasp. 50 g of BK-PP311 solid microbial inoculum is uniformly mixed and then placed in aerobic fermentation equipment with good heat preservation performance for aerobic fermentation, and uninterrupted ventilation and periodic stirring are needed in the process to ensure that the temperature is uniform and the aerobic fermentation is carried out. Meanwhile, a control group is arranged, and the same food waste and wheat bran are adopted for aerobic fermentation without being mixed with a solid microbial inoculum. 9 parts per day: 00 measuring the temperature of the center of the material, and simultaneously sampling and measuring various indexes such as the water content of the material. The moisture content of the materials and the temperature change of the compost body in the composting process are shown in figures 2 and 3.
The results show that Pantoea was added to the experimental groupsPantoeasp. After the BK-PP311 solid microbial inoculum, the temperature of a pile rapidly rises, the highest temperature is 75.0 ℃, the time required for reaching the highest temperature is 8d, the time required for reaching the highest temperature is 11d for a control group, and the time for reaching the highest temperature for an experimental group is shortened compared with that for reaching the highest temperature for the control group3d is added; in the composting process, the maintaining time of the experimental group in the high-temperature (55 ℃ and above) period is 18 d, the maintaining time of the control group in the high-temperature period is 13d, and the maintaining time of the experimental group in the high-temperature period is prolonged by 5 d compared with that of the control group in the high-temperature period. After composting is finished, the seed germination index of the experimental group fertilizer reaches 91.8 percent, and the number of faecal coliform (GB/T19524.1 for reference of the determination method) and the death rate of roundworm eggs (GB/T19524.2 for reference of the determination method) are respectively 95/g and 96.1 percent; the seed germination index of the control group fertilizer is 83.3%, and the number of fecal coliform and the death rate of roundworm eggs are 230/g and 63.8% respectively. Organic matter of experimental group fertilizer products, N, P2O5And K2The O content (the determination method is referred to NY 525-2O5The content (in terms of a dried basis) of (C) is 3.5%, K2The O content (calculated on a drying basis) is 2.7 percent; the organic matter content (calculated by a drying base) of the fertilizer product of the control group is 65%, the N content (calculated by the drying base) is 2.4%, and P is2O5The content (in terms of a dry basis) of (A) is 2.8%, K2The O content (on an oven-dried basis) was 2.4%. Thus, Pantoea was inoculatedPantoeasp.BK-PP311 can realize high-temperature aerobic fast composting of food waste and wheat bran, and improve the product quality of the fertilizer.
Example 4: high-temperature aerobic composting effect of strain BK-PP311 on fruit and vegetable garbage and pig manure as raw materials
Taking 5 kg of fruit and vegetable garbage and 5 kg of pig manure, uniformly mixing to obtain 10 kg of mixed material, and adding Pantoea according to the mass ratio of 0.2%Pantoeasp. 20 g of BK-PP311 solid microbial inoculum is uniformly mixed and then placed in aerobic fermentation equipment with good heat preservation performance for aerobic fermentation, and uninterrupted ventilation and periodic stirring are needed in the process to ensure that the temperature is uniform and the fermentation is carried out under aerobic conditions. Meanwhile, a control group is arranged, and the same fruit and vegetable garbage and pig manure are adopted for aerobic fermentation. 9 parts per day: 00 measuring the temperature of the center of the material, and simultaneously sampling and measuring various indexes such as the water content of the material. The moisture content of the materials and the temperature change of the compost body in the composting process are shown in figures 4 and 5.
The results show that Pantoea was added to the experimental groupsPantoeasp. After the BK-PP311 solid microbial inoculum, the temperature of a stack body is rapidly increased, the highest temperature is 70.4 ℃, the time required for reaching the highest temperature is 6 d, the time required for reaching the highest temperature is 8d for a control group, and the time for reaching the highest temperature for an experimental group is shortened by 2 d compared with the time for reaching the highest temperature for the control group; in the composting process, the high-temperature period maintaining time of the experimental group is 16 d, the high-temperature period maintaining time of the control group is 13d, and the high-temperature period maintaining time of the experimental group is prolonged by 3d compared with that of the control group. After composting is finished, the seed germination index of the experimental group fertilizer reaches 91.4 percent, and the number of faecal coliform bacteria and the death rate of roundworm eggs are respectively 98/g and 90.5 percent; the seed germination index of the control group fertilizer is 80.5%, and the number of fecal coliform and the death rate of roundworm eggs are 180/g and 61.2% respectively. Organic matter of experimental group fertilizer products, N, P2O5And K2The O content is higher than that of the control group, specifically, the organic matter content (calculated by a drying base) of the fertilizer product of the experimental group is 76 percent, the N content (calculated by the drying base) is 2.7 percent, and P is2O5The content (in terms of a dry basis) of (C) is 3.1%, K2The O content (calculated on a drying basis) is 2.8 percent; the organic matter content (calculated by a drying base) of the fertilizer product of the control group is 68 percent, the N content (calculated by the drying base) is 2.3 percent, and the P content is2O5The content (in terms of a dry basis) of (A) is 2.7%, K2The O content (on an oven-dried basis) was 2.3%. Thus, Pantoea was inoculatedPantoeasp.BK-PP311 can realize high-temperature aerobic rapid composting of fruit and vegetable garbage and pig manure, and improve the product quality of the fertilizer.
The strain organic solid waste high-temperature aerobic composting experimental result shows that the strain can realize the rapid composting of organic solid waste under aerobic condition, achieves the purposes of harmlessness, stabilization and recycling, can be used for treating organic solid waste such as food waste, kitchen waste, fruit and vegetable waste, livestock and poultry breeding manure and the like, and has high application value.
The above examples are intended to illustrate the disclosed embodiments of the invention and are not to be construed as limiting the invention. In addition, various modifications of the invention set forth herein, as well as variations of the methods of the invention, will be apparent to persons skilled in the art without departing from the scope and spirit of the invention. While the invention has been specifically described in connection with various specific preferred embodiments thereof, it should be understood that the invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the above-described embodiments which are obvious to those skilled in the art to which the invention pertains are intended to be covered by the scope of the present invention.
Sequence listing
<110> Shanghai higher research institute of Chinese academy of sciences
<120> organic solid waste high-temperature aerobic composting strain and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1419
<212> DNA
<213> Pantoea (Pantoea sp.BK-PP311)
<400> 1
gtggctcaaa aaggttaccc caccgacttc gggtgttaca aactctcgtg gtgtgacggg 60
cggtgtgtac aaggcccggg aacgtattca ccgcggcatg ctgatccgcg attactagcg 120
attccagctt catgtaggcg agttgcagcc tacaatccga actgagaacg gttttatgag 180
attagctcca cctcgcggtc ttgcagctct ttgtaccgtc cattgtagca cgtgtgtagc 240
ccaggtcata aggggcatga tgatttgacg tcatccccac cttcctccgg tttgtcaccg 300
gcagtcacct tagagtgccc aacttaatga tggcaactaa gatcaagggt tgcgctcgtt 360
gcgggactta acccaacatc tcacgacacg agctgacgac aaccatgcac cacctgtcac 420
tctgctcccg aaggagaagc cctatctcta gggttttcag aggatgtcaa gacctggtaa 480
ggttcttcgc gttgcttcga attaaaccac atgctccacc gcttgtgcgg gcccccgtca 540
attcctttga gtttcagcct tgcggccgta ctccccaggc ggagtgctta atgcgttaac 600
ttcagcacta aagggcggaa accctctaac acttagcact catcgtttac ggcgtggact 660
accagggtat ctaatcctgt ttgctcccca cgctttcgcg cctcagtgtc agttacagac 720
cagaaagtcg ccttcgccac tggtgttcct ccatatctct acgcatttca ccgctacaca 780
tggaattcca ctttcctctt ctgcactcaa gtctcccagt ttccaatgac cctccacggt 840
tgagccgtgg gctttcacat cagacttaag aaaccacctg cgcgcgcttt acgcccaata 900
attccggata acgcttgcca cctacgtatt accgcggctg ctggcacgta gttagccgtg 960
gctttctggt taggtaccgt caaggtgcca gcttattcaa ctagcacttg ttcttcccta 1020
acaacagagt tttacgaccc gaaagccttc atcactcacg cggcgttgct ccgtcagact 1080
ttcgtccatt gcggaagatt ccctactgct gcctcccgta ggagtctggg ccgtgtctca 1140
gtcccagtgt ggccgatcac cctctcaggt cggctacgca tcgttgcctt ggtgagccgt 1200
tacctcacca actagctaat gcgacgcggg tccatccata agtgacagcc gaagccgcct 1260
ttcaatttcg aaccatgcag ttcaaaatgt tatccggtat tagccccggt ttcccggagt 1320
tatcccagtc ttatgggcag gttacccacg tgttactcac ccgtccgccg ctaacttcat 1380
aagagcaagc tcttaatcca ttcgctcgac ttgcaaggt 1419

Claims (11)

1. Pantoea genusPantoea sp.) BK-PP311, with the preservation number of CCTCC NO: m2020973.
2. Pantoea (b) according to claim 1Pantoea sp.) BK-PP311, characterized in that said Pantoea genus (b), (c), (d), (Pantoea sp.) BK-PP311 comprises the gene sequence shown as SEQ ID NO. 1.
3. Pantoea (b) according to claim 1Pantoea sp.) BK-PP311, characterized in that said Pantoea genus (b), (c), (d), (Pantoea sp.) BK-PP311 withstands high temperatures of 75 ℃.
4. AA liquid microbial agent comprising the genus Pantoea (B) according to any one of claims 1 to 3Pantoea sp.) BK-PP311, of the genus Pantoea (b), (c) and (d) in the liquid inoculumPantoea sp.) BK-PP311 at a concentration of at least 1X 108 cfu/mL。
5. The method for preparing a liquid microbial inoculum according to claim 4, comprising the steps of: pantoea genus according to any one of claims 1 to 3 (b), (c), (d), (Pantoea sp.) BK-PP311 pure strains are inoculated in a liquid culture medium for culture, and after the culture is finished, the liquid microbial inoculum is obtained.
6. A solid bacterial agent comprising the genus Pantoea (B) according to any one of claims 1 to 3Pantoea sp.) BK-PP311 with a carrier.
7. The solid microbial inoculum according to claim 6, wherein the carrier is a solid phase carrier.
8. The method for preparing a solid microbial inoculum according to claim 6 or 7, which comprises the following steps: mixing a carrier and the liquid microbial inoculum according to the mass ratio of 1: 1-1: 10 of claim 4, and drying to obtain the solid microbial inoculum.
9. Pantoea (b) according to any one of claims 1 to 3Pantoea sp.) BK-PP311 is applied to organic solid waste aerobic composting.
10. The use of claim 9, wherein the organic solid waste comprises food waste, kitchen waste, fruit and vegetable waste, livestock and poultry breeding manure, municipal sludge.
11. A method for organic solid waste aerobic composting fermentation is characterized by comprising the following steps: mixing the solid microbial inoculum of claim 6 or 7 with organic solid waste materials, and then carrying out aerobic composting fermentation.
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