CN105567609A - High-temperature-resistant garden waste decomposing bacteria ST2 and application thereof - Google Patents

High-temperature-resistant garden waste decomposing bacteria ST2 and application thereof Download PDF

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CN105567609A
CN105567609A CN201610119770.7A CN201610119770A CN105567609A CN 105567609 A CN105567609 A CN 105567609A CN 201610119770 A CN201610119770 A CN 201610119770A CN 105567609 A CN105567609 A CN 105567609A
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temperature
composting
decomposing agent
garden waste
bacterium liquid
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CN105567609B (en
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彭霞薇
连鹏
周金星
郑景明
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Beijing Forestry University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/465—Streptomyces
    • 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
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • 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
    • 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
    • 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
    • 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 discloses a high-temperature-resistant cellulose degradation bacterium which is a streptomyces thermodiastaticus. The preservation number of the cellulose degradation bacterium is CGMCC No.12134. The bacterium is capable of producing a large quantity of enzymes at the high temperature of 40-70 DEG C; the cellulase of the bacterium is high in activity and has the characteristics of high temperature resistance and efficient degradation of cellulose; the bacterium can be used as a microbial agent, can be applied to a high-temperature composting system, and is suitable for composting the garden wastes.

Description

One strain high temperature resistant garden waste decomposer ST2 and application thereof
Technical field
The invention belongs to field of environmental biotechnology, be specifically related to the cellulosic thermoduric bacteria of a high-efficiency degradation that filters out from garden waste compost and the application in garden waste During High-Temperature Composting thereof.
Background technology
Garden waste refer to ornamental plant naturally wither and fall or manually prune produce deadwood, fallen leaves, grass bits, residual flower, trees and shrub beta pruning and other plant residue etc., main component is Mierocrystalline cellulose and the hemicellulose of difficult degradation.In recent years, the speed increase of the annual 8%-10% of China's garden waste, bring large drag forces to City Green process, the mode of current China process garden waste is mainly burns and fills up, and these two kinds of processing modes not only waste renewable resources and also create serious environmental pollution.Therefore, how rationally effectively to process garden waste, make its recycling be one of hot issue of everybody common concern at present.
Utilize composting technology garden waste to be become thoroughly decomposed and make organic fertilizer and seedling medium etc., it is one of effective outlet of its recycling, but the material containing a large amount of difficult degradations such as xylogen, Mierocrystalline cellulose in garden waste, cause that composting efficiency is low, the cycle is long, greatly limit the recycle value of garden waste.Therefore need to add specific microorganism in compost, to accelerate garden waste digest process.Cellulose-degrading bacteria is that a class can produce born of the same parents' outer fiber element enzyme and cellulose macromolecule is hydrolyzed into the microorganism of glucose, can the degradation speed of accelerating fibers cellulosic material.Occurring in nature, produces cellulosic microbe species a lot, and it is fungi that current investigation and application obtains more, as mould in wood, Penicillium, Aspergillus, Rhizopus etc.Bacterium and actinomycetic investigation and application all less.In composting process, cellulose degradation mainly occurs in the pliotherm period, and fungi is main under mesophilic condition, enzyme is lived maximum, along with the rising of composting process temperature, the enzymic activity of fungi number of viable and generation thereof reduces greatly, which limits the utilization in compost of cellulase-producing mould.Screening and the application of high-temperature fibre element degradation bacteria are the effective measure solved the problem.
A lot of cellulose-degrading bacteria is all for agricultural wastes such as maize straw, straw, wheat straws, seldom has specially for screening and the research on utilization of the degradation bacteria of garden waste cellulose materials.Chinese patent " plant height temperature cellulose-degrading bacteria and an application thereof " (number of patent application: 201410018582.6) disclose the high temperature fiber element degradation bacteria ground bacillus that a strain is separated from garden waste pliotherm period compost sample, cellulase activity is 7.8U/ml, and this bacterium is bacterium.About being separated from garden waste compost, the actinomycetic report of high-temperature fibre element degraded is less.
Summary of the invention
Technical problem to be solved by this invention is: provide a strain high temperature resistant thermophilic streptomyces diastaticus, this bacterium can in 40-70 DEG C of high temperature large volume production enzyme, cellulase activity is high, there is high temperature resistant, efficient degradation cellulosic nature, microbiobacterial agent can be it can be used as to be applied in During High-Temperature Composting system, to be applicable to garden waste compost.
Technical scheme provided by the invention is: the thermophilic streptomyces diastaticus of a strain high-temperature fibre element degradation bacteria ( streptomycesthermodiastaticus) ST2, deposit number is CGMCCNo.12134, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center.
Thermophilic streptomyces diastaticus of the present invention ( streptomycesthermodiastaticus) on February 18th, 2016, for the preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC institute, (preservation address was ST2: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode: 100101), its preserving number is CGMCCNo.12134, survives after testing.
The present invention is thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus) ST2, be the bacterial classification be separated from the sample of apple orchard, Changping County, Beijing district garden waste compost high temperature collection in mid-term, be numbered bacterial strain ST2, this bacterium can produce cellulase and degraded cellulose under 40-70 DEG C of condition.When bacterial strain grows on Gao Shi I substratum, aerial hyphae is light grey, and mycelia multiple-limb, substrate mycelium lark, does not produce soluble pigment, and aerial hyphae branch, aerial hyphae has fibrillae of spores.Under microscope, thalline is mycelioid, and gramstaining is positive, spore oval, smooth surface, and fibrillae of spores is slightly bending, in the shape of a spiral.In conjunction with bacterium colony morphological features and the Phylogenetic Analysis based on bacterial 16 S rDNA gene order, be accredited as thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus).
The method of bacterial strain production of cellulose enzyme of the present invention, is inoculated in fermention medium (medium component: CMC-Na0.5%, peptone 1%, wheat bran 3%, NaCl2%, KH by this bacterium 2pO 40.1%g, MgSO 47H 2o0.02%, (NH 4) 2sO 40.3%, pH7.0-7.6) in, turn in shaking table in temperature 50 C, rpm150 and cultivate 3-4d, centrifuging and taking supernatant liquor, measure cellulase activity.
The present invention also provides a kind of and is applicable to the During High-Temperature Composting decomposing agent that garden waste is main composting material, and this decomposing agent obtains in liquid fermenting mode, and bacterial concentration is 5 × 10 9cFU/mL, wheat bran and Semen Maydis powder 2:1 mix as bacterium liquid sorbent material, mix, be made into solid-state microbial inoculum by bacterium liquid with sorbent material 1:1.
Simultaneously, the present invention also provides the application of described thermophilic streptomyces diastaticus ST2 bacterial strain in the compost maturity taking garden waste as main raw, be main composting material with garden waste, add animal excrement and water, mixture carbon-nitrogen ratio is made to be 25-40:1, water ratio 50-60%, solid-state decomposing agent is inoculated in compost material in 5% ratio of weight of material, carries out During High-Temperature Composting.
The present invention has following beneficial effect:
Thermophilic streptomyces diastaticus of the present invention ( streptomycesthermodiastaticus) ST2 is the bacterial classification filtering out degraded cellulose from garden waste compost, can produce more cellulase in 40-70 DEG C of temperature range, enzyme work, up to 63.55U/mL, has high temperature resistant, efficient degradation cellulosic nature.Relative fungi under mesophilic condition enzyme lives maximum, and the problem that Production by Bacteria enzyme activity is lower, thermophilic streptomyces diastaticus can adapt to hot environment, higher cellulase can be produced again, thermophilic streptomyces diastaticus to cellulose materials, there is good degradation capability, so more can adapt to composting process complex environment.
By the present invention obtain high-temperature fibre element degradation bacteria make solid-state decomposing agent add to garden waste be main raw compost in, compared with the contrast do not inoculated, time, prolongation time length pliotherm period pliotherm period temperature that compost enters the pliotherm period can be improved, reduce composting C/N ratio, thus accelerate compost maturity process.Microbiobacterial agent can be it can be used as to be applied in During High-Temperature Composting system, to be applicable to garden waste compost.
The present invention be directed to the bacterial classification of garden waste composting material screening and the decomposing agent of preparation, reasonable, an effective approach can be provided for the recycling of garden waste, realize the object reducing environmental pollution, resource circulation utilization.
Accompanying drawing explanation
Fig. 1 is thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus) separation and purification picture
Fig. 2 the present invention is thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus) 16SrDNA gene order.
Fig. 3 by thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus) and the gene order of close bacterial strain 16SrDNA carries out homology Blast comparison time the phylogenetic tree picture that builds.
Heap temperature change in Fig. 4 composting process.
Carbon-nitrogen ratio (C/N ratio) change in Fig. 5 composting process.
Embodiment
Detailed description below by embodiment illustrates the present invention further, but is not limitation of the present invention, only does example explanation.
the screening of embodiment 1 high-temperature fibre element degradation bacteria strains
Collected specimens from apple orchard, Changping County, Beijing district garden waste compost high temperature initial stage, high temperature mid-term, high temperature post material, Yanqing County of Beijing Plain afforestation afforestation waste stores; Take above-mentioned fresh sample 10g to be put in and 10 granulated glass spherees to be housed and to fill in the Erlenmeyer flask of 90ml sterilized water, the shaking table being placed in 30 DEG C of 150rpm shakes 30min, and sample is fully scattered, and leaves standstill enrichment culture 24h at 50 DEG C.Draw 1ml supernatant liquor with aseptic straw to join in the test tube containing 9ml sterilized water, this is 10 -1sample diluting liquid, then from 10 -1get 1ml in sample to join in 9ml sterilized water, this is 10 -2sample diluting liquid, by that analogy, obtains 10 -3, 10 -4, 10 -5, 10 -6sample diluting liquid, then draws 10 of 100 μ l with pipettor -3, 10 -4, 10 -5, 10 -6on Cellulose and congo red differential medium, (substratum consists of sample diluting liquid: K 2hPO 40.5g, Microcrystalline Cellulose 1.88g, MgSO 40.25g, gelatin 2.0g, Congo red 0.5g, agar 16g, distilled water 1000ml, pH7.0), with spreader, diluent is spread evenly across whole flat board, and is placed in 50 DEG C of incubators cultivations 3 days.Select the bacterium colony having obvious transparent circle on the Congo red flat board of Mierocrystalline cellulose, be numbered, then separation and purification of repeatedly ruling obtains purebred bacterial strain, receive on inclined-plane by the bacterial classification after being separated, subsequent experimental is carried out in 4 DEG C of preservations.
The pure bacterial strain being preserved in 4 DEG C is transferred to carboxymethyl cellulose substratum (medium component: CMC-Na15.0g, NH 4nO 31.0g, yeast extract paste 1.0g, MgSO 47H 2o0.5g, KH 2pO 41.0g, distilled water 1000ml, agar 16g, pH7.0) on flat board, activation culture at 50 DEG C, single bacterium colony of then provoking on flat board is transferred on the Congo red flat board of Mierocrystalline cellulose, is placed in 50 DEG C of incubators and cultivates, and measures colony diameter d and transparent circle diameter D after 72h, calculate its ratio H, namely H=D/d, H value is larger, and it is stronger to be worth the cellulolytic ability of this bacterial strain of larger expression.Tentatively determine that its cellulase-producing is active according to the size Congo red qualification substratum of Mierocrystalline cellulose being formed transparent circle.
Pass through aforesaid operations, obtain many strains cellulose-degrading bacteria, the bacterial classification be wherein separated in the sample of apple orchard, Changping County, Beijing district garden waste compost high temperature collection in mid-term, called after ST1's, China Committee for Culture Collection of Microorganisms's common micro-organisms center is deposited on February 15th, 2016, it is referred to as CGMCC(unit address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101), deposit number is CGMCCNo.12134.Carry out optical microscope and gramstaining qualification to ST2 bacterial strain, when should grow on Gao Shi I substratum, aerial hyphae is light grey, mycelia multiple-limb, substrate mycelium lark, does not produce soluble pigment, aerial hyphae branch, aerial hyphae has fibrillae of spores.Under microscope, thalline is mycelioid, and gramstaining is positive, spore oval, smooth surface, and fibrillae of spores is slightly bending, in the shape of a spiral, sees Fig. 1.
embodiment 2ST2 bacterial strain molecular biology identification
Molecular Identification is carried out to screening the thermophilic streptomyces diastaticus obtained, carry out according to following steps: single colony inoculation of picking bacterium is in liquid Gao Shi I substratum, 30 DEG C, 120r/min shaking table shaking culture, nutrient solution is taken out at 2d, 5000r/min is centrifugal, and 1min gets supernatant liquor, according to bacterial genomes DNA extraction kit (Tian Gen biochemical technology company limited provides), extract bacterium colony DNA; Universal primer 27F and 1492R carries out pcr amplification to the DNA of bacteria extracted; 27F sequence is 5 '-AGAGTTTGATCCTGGCTCAG-3 '; 1492R sequence is 5 '-AAGGAGGTGATCCAGCCGCA-3 '; PCR primer is carried out sequence, and sequencing result BLAST in ncbi database carries out sequential analysis, and carries out tetraploid rice.
16SrDNA gene order (the referring to Fig. 2) length of Potsdam bacillus brevis is 1426bp, gene order is submitted on Genbank, carries out tetraploid rice, then adopt MEGA6.0 Software on Drawing phylogenetic tree, see Fig. 3, thus determine the kind of bacterial strain.Result show this sequence and thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus) 16SrDNA gene order similarity up to 99%, simultaneously in conjunction with colony morphology characteristic, physiological and biochemical property, thalline microscopic features determine ST1 bacterial strain be thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus).
embodiment 3 is thermophilic streptomyces diastaticus growth measurement
By thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus) ST2 is inoculated into (CMC-Na15.0g, NH in CMC liquid nutrient medium 4nO 31.0g, yeast extract paste 1.0g, MgSO 40.5g, KH 2pO 41.0g, distilled water 1000mL), get nutrient solution every 2h, connection is continuous is sampled to 48h, and measuring the OD600 value in each period, take incubation time as X-coordinate, and the OD600 value of each sampling spot is ordinate zou, draws the growth curve of this bacterium, i.e. the growth measurement of this bacterium.Can find out that 0-8h is lag period, 9 ~ 16h is logarithmic phase, 16 ~ 24h is stationary phase, >26h is decline phase from measurement result.The growth of the bacterial strain of logarithmic phase is rapid, energetic, in therefore later enzymatic production experiment, the fermented liquid of 12 hours should be selected to be that seed liquor is inoculated.
embodiment 4 is thermophilic streptomyces diastaticus cellulase-producing vitality test
By thermophilic for logarithmic phase streptomyces diastaticus ST2 by 1% inoculum size be inoculated into fermention medium (medium component: CMC-Na0.5%, peptone 1%, wheat bran 3%, NaCl2%, KH 2pO 40.1%, MgSO 47H 2o0.02%, (NH 4) 2sO 40.3%), pH7.0-7.6) in, turn in shaking table in temperature 50 C, rpm150 and cultivate 3-4d, by centrifugal for culture 8000r/min 5min, get supernatant and be crude enzyme liquid, measure cellulase activity by DNS method.Get 1mL supernatant liquor (replacement of blank 1ml distilled water) in test tube, be placed in 50 DEG C of water-baths, preheating 2min, then add 4ml at the substrate solution of 50 DEG C of preheatings, take out after clock reaction 5min, add 4mLDNS nitrite ion, be positioned in boiling water bath after shaking up and heat 5min, put into cooling bath after taking-up immediately to cool, be settled to 20ml with distilled water, after mixing, measure absorbancy at 540nm wavelength place.Convert after reference standard curve the product enzyme activity of this bacterial strain.Enzyme activity unit is according to international unit stipulative definition, and namely in 1mL system, in 1min, catalyzing cellulose hydrolysis generates the enzyme amount needed for 1 μm of ol glucose is an enzyme activity unit (U/mL).The cellulase activity of ST2 bacterial strain is 63.55U/mL after measured.
the preparation of the solid-state decomposing agent of embodiment 5
(1) bacterial strain activation: get 4 DEG C, microorganism of the present invention and preserve inclined plane inoculating Zhi Gaoshi I solid plate substratum, cultivate 12h and realize bacterial strain activation in the permanent case of 50 DEG C.
(2) seed liquor preparation: be seeded in 1L aseptic Gao Shi I liquid nutrient medium through dull and stereotyped 1 of the bacterial classification of slant activation by step (1), obtains seed liquor after cultivating 12h under 50 DEG C of 150rpm shaking table conditions.
(3) preparation of fermentation seed liquid: above-mentioned seed liquor is by 6-10%(v/v) inoculum size be seeded in sterilized fermentor tank, carry out expansion fermentation culture.Under the condition of temperature 50 C, oscillation frequency 120rpm, after cultivating 48h, obtain fermentation seed liquid.
(4) preparation of solid-state decomposing agent: using after wheat bran mixes according to 2:1 mass ratio with Semen Maydis powder as bacterium liquid sorbent material, bacterium liquid with obtained in step (3), mixes with sorbent material volume mass ratio 1:1 by bacterium liquid, is made into solid-state decomposing agent, after banking up 1 week, can be used for During High-Temperature Composting.
the compost effect test of embodiment 6 decomposing agent
Be main composting material with garden waste, add animal excrement and water, make mixture carbon-nitrogen ratio be 25-40:1, water ratio 50-60%, solid-state decomposing agent is inoculated in compost material in 5% ratio of weight of material, carries out During High-Temperature Composting, not add the material of decomposing agent for contrast, when heap temperature rises to 50 DEG C, start turning, the turning of every 2 days of pliotherm period once, cooldown period weekly turning once, not in turning after temperature drops to 40 DEG C.In composting process, by measuring temperature variation, the change of composting material carbon nitrogen (C/N) ratio of heap body every day, investigating and adding decomposing agent to the impact of garden-waste compost decomposition progress.
In composting process, heap temperature change as shown in Figure 4.As shown in Figure 4, the compost treatment of inoculation decomposing agent rises to more than 50 DEG C in compost 2d temperature, and the pliotherm period of more than 50 DEG C continues 25d, and temperature starts to decline afterwards.And nonvaccinated compost treatment temperature just rises to more than 50 DEG C at compost 3d, postpone 1d than the process of inoculation and arrive more than 50 DEG C pliotherm periods, and more than 50 DEG C time length pliotherm period is 20d, 5d fewer than the process of inoculation, the top temperature of the heap body pliotherm period of inoculation process is also higher than not inoculating process, compost can be accelerated after inoculation decomposing agent is described and enter time pliotherm period, and time length pliotherm period.In composting process, C/N change as shown in Figure 5.As shown in Figure 5, along with the carrying out of compost, inoculation decomposing agent and the process C/N do not inoculated reduce than all continuing, and the process decline degree of inoculation is higher than not inoculating process, illustrate thus, the decomposing agent adding this bacterium obtained can accelerate composting process, improves the effect of becoming thoroughly decomposed of garden waste compost.

Claims (5)

1. a strain high-temperature fibre element degradation bacteria thermophilic streptomyces diastaticus ( streptomycesthermodiastaticus), preserving number is CGMCCNo.12134.
2. one kind is applicable to the During High-Temperature Composting decomposing agent that garden waste is main composting material, it is characterized in that: this decomposing agent obtains in liquid fermenting mode, its raw material is bacterium liquid and sorbent material, be mixed in proportion by bacterium liquid and sorbent material, be made into solid-state microbial inoculum, wherein bacterium liquid be thermophilic streptomyces diastaticus according to claim 1 ( streptomycesthermodiastaticus) bacterium liquid, sorbent material is wheat bran and Semen Maydis powder.
3. During High-Temperature Composting decomposing agent as claimed in claim 2, is characterized in that: described bacterial concentration is 5 × 10 9cFU/mL, wheat bran and Semen Maydis powder 2:1 mix as bacterium liquid sorbent material, mix, be made into solid-state microbial inoculum by bacterium liquid with sorbent material 1:1.
4. the application of During High-Temperature Composting decomposing agent in compost maturity as described in Claims 2 or 3.
5. apply as claimed in claim 4, it is characterized in that: be main composting material with garden waste, add animal excrement and water, mixture carbon-nitrogen ratio is made to be 25-40:1, water ratio 50-60%, solid-state decomposing agent is inoculated in compost material in the 2.5%-5% ratio of weight of material, carries out During High-Temperature Composting.
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CN106350075A (en) * 2016-08-28 2017-01-25 何晓东 Soil improvement agent with sterilizing effect and preparation method of soil improvement agent
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CN114507090A (en) * 2021-12-28 2022-05-17 镇江贝思特有机活性肥料有限公司 Tobacco stem waste fermentation treatment device and treatment method
CN115197023A (en) * 2022-05-30 2022-10-18 北京林业大学 Bio-organic fertilizer with landscaping waste as matrix and preparation method thereof

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106350075A (en) * 2016-08-28 2017-01-25 何晓东 Soil improvement agent with sterilizing effect and preparation method of soil improvement agent
CN107760616A (en) * 2017-07-25 2018-03-06 青岛海芬健康产业科技有限公司 A kind of microbial bacterial agent for rubbish from cooking of degrading and preparation method thereof
CN114507090A (en) * 2021-12-28 2022-05-17 镇江贝思特有机活性肥料有限公司 Tobacco stem waste fermentation treatment device and treatment method
CN115197023A (en) * 2022-05-30 2022-10-18 北京林业大学 Bio-organic fertilizer with landscaping waste as matrix and preparation method thereof
CN115197023B (en) * 2022-05-30 2024-04-19 北京林业大学 Bio-organic fertilizer with landscaping waste as matrix and preparation method thereof

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