CN109943508A - A kind of microbial germ powder evaluation method promoting garden waste degradation-type - Google Patents
A kind of microbial germ powder evaluation method promoting garden waste degradation-type Download PDFInfo
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Abstract
The present invention discloses a kind of microbial germ powder evaluation method for promoting garden waste degradation-type, it is to afforestation waste microbe inoculation bacterium powder, it is placed in 30 ± 1 DEG C of constant temperature incubations, the quality change situation of results of regular determination afforestation waste degradation residue evaluates microbial germ powder to the degradation efficiency of afforestation waste by degradation weight-loss ratio；When cultivating 15d, microbial germ powder is not less than 30% to the degradation weight-loss ratio of afforestation waste；The microbial germ powder is by Cellumomonas flavigena thallus and adsorptive support, protective agent, with mass ratio 1:(0.5-1.5): (0.5-1.5) is mixed.Adsorptive support selects dextrin, diatomite, talcum powder or calcium carbonate；Wetting agent selects Tween 80, polyethylene glycol, soluble starch or lauryl sodium sulfate；Protective agent selects turf, kaolin, sodium alginate or sodium cellulose glycolate.Microbial germ powder of the invention is used for afforestation offal treatment, can meet the needs of scale compost disposition afforestation waste.
Description
Technical field
The invention belongs to the efficient technique of rainwater utilization field of agricultural organic resource, it is related at afforestation waste degradation
A kind of microbial bacterial agent set, and in particular to microbial germ powder applicating evaluating method for promoting afforestation waste degradation.
Background technique
In recent years, with the beautiful Chinese in-depth of theory and the expansion of Construction of Eco-urban scale, China's urban afforestation water
Flat, whole green amount is continuously improved, while also producing a large amount of afforestation wastes.City trees and shrubs waste is mainly
Refer to the lawn generated in green plants nature or maintenance processes in urban green space, shade tree and outskirts of a town forest land, dry branches and fallen leaves and
Forest lop etc..These city trees and shrubs wastes contain a large amount of starch, cellulose, lignin and nutrient, are a kind of
The important component of recycling organic resource and ecosystem substance circulation.However, as urban solid garbage
One of main source has nearly 50,000,000 tons of garden waste to enter environmental sanitation landfill system every year, not only causes gardens biological
The matter wasting of resources and refuse landfill storage ability decline, also easily lead to the secondary pollution of surrounding water, atmosphere and soil environment.
The resource utilization mode of city trees and shrubs waste mainly crushes on the spot and is used as green soil covering or simple
Compost exists as plant growth medium and crushes the problems such as being also easy to produce the not thorough easily twin germ of fugitive dust, compost and foul smell.And
By centainly pre-processing, under optimum conditions using aerobic microbiological carry out compost fermentation, promote larger molecular organics degradation and
It is to realize afforestation changing rejected material to useful resource, the efficient main path utilized that humus material, which is formed,.Studies have shown that fiber
Plain enzyme is that catalytic cellulose fast hydrolyzing and induction lignin-degrading enzymes (extracellular peroxidase and extracellular phenol oxidase) generate
Core enzyme system, mainly includes endoglucanase, excision enzyme and glycosides enzyme, and cellulose degradation is cellooligosaccharide by synergistic effect
And glucose.Therefore, fast degradation how is constructed, the thoroughly decomposed and convenient microbial bacterial agent of application is to promote gardens cellulose
The key that equal macromolecular substances decompose, convert, synthesize the soil organism and plant growth nutriment.
Currently, the domestic microbial bacterial agent relation technological researching about degradation agriculture and forestry organic waste material cellulose be concentrated mainly on
Under several aspects:
(1) straw decomposing microbial inoculum
Cellulose is the main material composition ingredient of stalk, because its chemical structure is complicated, property is stablized, while stalk surface
Water-insoluble wax coat is easily formed, causes its natural decomposition speed slow.Separately since the C/N of stalk is than excessively high, direct returning to farmland meeting
There is the phenomenon that edaphon and crop contention nitrogen source, to influence seedling growth and crop yield.Therefore, fast prompt drop is developed
The effective technology means that solution, efficiently decomposed stalk microbe microbial inoculum are agricultural waste disposal of resources and utilize.Wang Yuanming
It screens and is prepared within (2013) to be suitable under hot conditions, the microbial bacterial agent of degradation and decomposed rice straw.It studies knot
Fruit are as follows: two kinds of high temperature strepto- category zymocyte liquids are mixed by equal proportion, are prepared into the decomposed bacterium solution of instant, high temperature aerobic composting 20
Decomposed effect of preferably degrading can be realized in its rice straw, and the percentage of seedgermination of decomposed product reaches 85.96% (Wang Yuan
It is bright, research of the screening and its composite bacteria agent of high temperature fiber element degradation bacteria to straw degradative effect, Agricultural University Of Nanjing, 2013).
This has many advantages, such as degradation rate height, decomposed thorough, adaptation compost hot environment using microbial inoculum, but the microbial inoculum is liquid form, is deposited
The problems such as being not easy preservation and Applicable temperature condition is limited to, therefore it is unable to satisfy technical grade, large-scale production demand.Yin Zhongwei
(2010) the microbial composite bacteria systems screened and be prepared for suitable for wheat stalk of degrading.Its result of study are as follows: in Wheat Straw
During stalk Liquid Culture, composite microbial system Y2b to the degradation rate of cellulose and hemicellulose up to 40% or more, and its cellulose
Enzyme activity can reach highest in 2d, be 61.2U/mL (Yin Zhongwei, the screening of stalk cellulose efficient degrading bacterial strain and to straw
Stalk degradation effect Primary Study, China Agricultural University, 2010).The composite microbial system has flora multiplicity, inulinase-producing activity height, degradation
The advantages that efficiency is significant, but its contained strain INFORMATION OF INCOMPLETE, microbial inoculum form are liquid, are only applicable to laboratory research analysis,
There may be the risks such as pathogen infection for industrial application.
(2) gardens biomass compost bacterium
The Composting treatment of gardens biomass keeps biological organic matter real under the action of microorganism by hot fermentation
The process of existing mineralization, humification and innoxious decomposed product, wherein microorganism (bacterial strain or fungus strain) can improve the physics and chemistry of windrow
Property accelerates decomposed, the rush conversion of compost to form humus.It is bent that Tian Yun (2012) has studied external source fermentation of organic wastes bacterium
The influence to gardens pruning object compost maturity effect is added with bamboo vinegar liquid ratio.Its result of study are as follows: to garden waste compost
Middle addition 0.5% fermentation of organic wastes bacterium song+1000 times of dilution bamboo vinegar liquid 2L, can effectively improve the rising of compost initial stage temperature
Amplitude accelerates compost maturity speed, promotes composting production quality, while the addition of certain extension rate bamboo vinegar liquid can effectively reduce
Nitrogen loss improves micro- bacteria-promoting agent activity (Tian Yun, the application study of garden waste Composting treatment and products thereof, Beijing woods
Sparetime university is learned, and 2012).It is discarded in compost disposition in gardens, the addition of zymophyte song and bamboo vinegar liquid can effectively improve composting production
Full nitrogen, full phosphorus, full potassium and iron and sulphur mass fraction, promote the survival rate of Calathea orbifolia cultivation, and improve its plant height with
Biomass.But the kind information of the biological effectiveness of bacterium song and wherein microorganism is simultaneously indefinite.It sieves within Shi Longxiang (2015)
Select and assembled the microbial flora of suitable degradation fruit tree cellulose.Its result of study are as follows: Penicillium notatum (Penicillium
Sp.), Acremonium chrysogenum (Acremonium alternatum), yellow grey Penicillium notatum (Penicillium aurantioqriseum)
Lead to bacterium (Ralstoinia sp.) equal proportion with Rolls and assemble gained liquid microbe flora, fruit tree heap can be effectively improved
The decomposition temperature of body extends the high temperature decomposition phase, promotes material decomposed, and cellulase, laccase, manganese peroxidase and lignin
The activity of peroxidase be significantly increased (Shi Longxiang, cellulose-degrading bacteria screening and its answering in fruit tree decomposition
With Xibei Univ. of Agricultural & Forest Science & Technology, 2015).The experiment sieving and assembled one group can efficient degradation fruit tree cellulose it is micro-
Biological fungus strain, wherein the cellulase activity of Penicillium notatum is apparently higher than combination fungus strain, and plays a significant role in the compost megathermal period.So
And the equal proportion that the fungus strain is still each bacterium solution assembles, which kind of bacterial strain plays a crucial role and how to participate in degradation process not
It is clear, while the biological effectiveness of liquid fungus strain is not also studied.
Summary of the invention
As urban afforestation level greatly improves, a large amount of afforestation waste generates therewith, however its biomass provides
The inefficient disposition in source is city management and the main problem that Green Development faces.Cellulose is mainization of city trees and shrubs waste
One studied point.How in urban landscaping the waste fast degradation of cellulose and gardens biomass resource efficient is realized
Using being the present patent application Key technique problem to be solved.
Present patent application inventor obtained by a large amount of full and accurate tests, Cellumomonas flavigena (Cellulomonas
Flavigena the cellulose that) can effectively degrade in urban landscaping waste.To highlight the Cellumomonas flavigena
(Cellulomonas flavigena) efficiently, the application advantage of low energy consumption processing afforestation waste, the present invention is further
Cellumomonas flavigena (Cellulomonas flavigena) is prepared into the microbial germ powder for facilitating storage and using, and is commented
Its effect in the disposition of afforestation waste degradation of valence.
Specifically, a kind of evaluation method of microbial germ powder, afforestation waste are inoculated with certain agent after physical treatment
The microbial germ powder of amount is placed in (30 ± 1 DEG C) of constant temperature in suitable environment temperature cultures, results of regular determination afforestation waste drop
The quality change situation for solving residue, by degrading, weight-loss ratio evaluates degradation of the microbial germ powder to afforestation waste
Efficiency；
When cultivating 15d, the microbial germ powder is not less than 30% to the degradation weight-loss ratio of afforestation waste；
The microbial germ powder the preparation method comprises the following steps: by Cellumomonas flavigena (Cellulomonas flavigena) bacterium
Body and adsorptive support are mixed with the ratio of quality 1:(0.5-1.5)；Add wetting agent, the Cellumomonas flavigena
The ratio of (Cellulomonas flavigena) thallus and wetting agent is 1:(0.5-1.5)；Protective agent is added, the production is yellow
Cellulomonas cartae (Cellulomonas flavigena) thallus and protectant ratio are 1:(0.5-1.5)；It stirs and evenly mixs, very
Sky is drying to obtain the microbial germ powder；
The adsorptive support selects dextrin, diatomite, talcum powder or calcium carbonate；
The wetting agent selects Tween 80, polyethylene glycol, soluble starch or lauryl sodium sulfate；
The protective agent selects turf, kaolin, sodium alginate or sodium cellulose glycolate.
The adsorptive support, wetting agent and protectant selection can effectively ensure to produce yellowish fiber list in microbial germ powder
The living bacteria count amount of born of the same parents bacterium.Verified, the key technical indexes of microbial germ powder of the present invention meets " agricultural microorganism
Microbial inoculum (GB 20287-2006) " in organic matter decomposing inoculant product pulvis index request.This microbial germ powder is stored up in 390d
It deposits in the phase, the viable count of Cellumomonas flavigena is not less than 0.5 × 108A/g, cellulase activity are not less than 30U/g.
The quality such as the microbial germ powder by above-mentioned preparation are sub-packed in brown Envelope bag, and sealing is closed, and room temperature (10-30 DEG C)
It deposits in medicine storage cabinet.Living bacteria count through different Storage periods (30d, 90d, 180d, 270d, 330d, 390d, 450d) bacterium powder
Show that in 390d Storage period, the key technical indexes meets " agricultural microbial agent (GB with cellulase activity measurement
20287-2006) " under item organic matter decomposing inoculant product pulvis index request, i.e. living bacteria count >=0.50 × 108A/g,
Cellulase activity >=30U/g.
As one of the embodiment of the specific microbial germ powder evaluation method of the invention, it is protected from light used in 10-30 DEG C
Store the microbial germ powder degradation afforestation waste of 390d, drop of the microbial germ powder to afforestation waste
It solves weight-loss ratio and is not less than 30%.
As a preferred embodiment, the afforestation waste is reed (Phragmites
Australis), tulip (Tulipa gesneriana), silver wire grass (Chloranthus japonicus), iris (Iris
Tectorum one or more of withered stalks).
Another level, the present invention give application of this microbial germ powder in degradation afforestation waste.As
The further explanation of the application, the microbial germ powder is with Cellumomonas flavigena (Cellulomonas flavigena) bacterium
Body is as active bacteria, for the cellulose in afforestation waste of degrading.
Compared with prior art, beneficial effects of the present invention or advantage major embodiment be in the following areas: the present invention is to produce Huang
Effective active microorganism of the cellulomonas cartae (Cellulomonas flavigena) as microbial germ powder, the microbial germ powder
The degradation of gardens cellulose can efficiently be promoted.In 390d Storage period, major technique refers to microbial germ powder prepared by the present invention
Reference symbol closes the index request of organic matter decomposing inoculant product pulvis in " agricultural microbial agent (GB 20287-2006) ", that is, has
Imitate viable count >=0.50 × 108A/g, cellulase activity >=30U/g.Using condition storage microbial germ powder of the present invention
When to 450d, 5 ‰ bacterium powder suspensions to reed stalk constant temperature incubation to 5d, 10d, 15d when degradation weight-loss ratio be respectively
16.8%, 22.2%, 29.6% (being averaged), opposite blank control group (CK:1.01%2.93%3.59%) are respectively increased
1563%, 657% and 724%.
Specific embodiment
Below with reference to embodiment, the present invention is further illustrated, and following each embodiments are merely to illustrate the present invention, right
The present invention is not restricted.
Embodiment 1
The present embodiment provides a kind of microbial germ powder of degradable gardens cellulose, effective viable bacteria is with extremely strong degradation
The Cellumomonas flavigena (Cellulomonas flavigena) of cellulose ability.Cellumomonas flavigena used in the present embodiment
Thallus is Cellulomonas type culture, and the strain purchase is from China Microbial Culture Preservation Commission's common micro-organisms
The heart.For the solid-like microbial germ powder for preparing shelf-stable, facilitating application, the present embodiment is by the liquid fermentation of Cellumomonas flavigena
Thallus is mixed with suitable adsorptive support, wetting agent and the protective agent of special ratios.Specifically, adsorptive support select dextrin,
One of diatomite, talcum powder or calcium carbonate are a variety of, and by quality ratio, the Cellumomonas flavigena body and adsorptivity carry
The ratio of body is 1:(0.5-1.5)；Wetting agent is selected in Tween 80, polyethylene glycol, soluble starch or lauryl sodium sulfate
It is one or more, by quality ratio, the ratio of the Cellumomonas flavigena body and wetting agent is 1:(0.5-1.5)；Protection
One of turf, kaolin, sodium alginate or sodium cellulose glycolate or a variety of are selected in agent, and by quality ratio, the production is yellow
Cellulomonas cartae body and protectant ratio are 1:(0.5-1.5).Further, the present embodiment will be combined and specifically be tested
Journey illustrates adsorptive support, wetting agent and protectant selection type and dosage.
The determination of adsorptive support type, by measuring the suction bacterium amount of different adsorption cycle used carriers, i.e. living bacteria count
As evaluation index.Specific measuring method are as follows: zymocyte liquid and adsorptive support are with the ratio of 10:1 (volume mass ratio, v/m)
Be suspended, stand 30min, be centrifuged 5min through 3500rpm revolving speed, remove surface sediments and be placed in 35 DEG C of incubators, respectively the 1st,
3, its living bacteria count is measured when 5h and the 1st, 3,5d, sort screening best adsorption from more to less according to living bacteria count quantity
Carrier.
The selection of different adsorptive supports and each adsorptive support different adsorption cycles living bacteria count referring to table 1.This
Experiment investigation dextrin, diatomite, talcum powder, calcium carbonate are as adsorptive support to the suction bacterium amount of Cellumomonas flavigena.From
Table 1 is as can be seen that dextrin, diatomite, talcum powder, calcium carbonate can effectively adsorb Cellumomonas flavigena, wherein dextrin exists
The living bacteria count amount that Cellumomonas flavigena is adsorbed when the 1st, 3,5h and the 1st, 3,5d is respectively (3.15-3.21) × 108A/
g、(3.06-3.12)×108A/g, more than diatomite, talcum powder, calcium carbonate corresponding period living bacteria count.Therefore,
Dextrin can be selected as preferred adsorptive support.
Living bacteria count of the different adsorptive supports of table 1 in different adsorption cycles
The determination of wetting agent type sinks to the bottom time as evaluation index using measuring graduates method measurement bacterium powder sample.
Specific evaluation method are as follows: screen the centrifugal deposition object of gained adsorptive support, according to the above method with 1:1's (mass ratio, m/m)
Ratio adds different type wetting agent, and for 24 hours, grind into powder weighs 1g sample and is quickly poured into and fills 500mL for 35 DEG C of vacuum drying
It in the 500mL measuring graduates of water, does not stir, at once manual time-keeping, 99% sample sinks to the bottom time and shorter shows wetability
Better.Different wetting agent corresponds to the sedimentation time of bacterium powder sample, referring to table 2.
2 different wetting agent of table corresponds to the sedimentation time of bacterium powder sample
The wetting of Tween 80, polyethylene glycol, soluble starch, lauryl sodium sulfate to bacterium powder sample of this experiment investigation
Performance.From table 2 it can be seen that Tween 80, polyethylene glycol, soluble starch, lauryl sodium sulfate have profit to bacterium powder sample
It is moist, but wetability each other has differences, wherein when polyethylene glycol sinks to a tin bottom to bacterium powder sample in 3 tests
Between measurement result be respectively 35s, 38s, 36s, the sedimentation time is significantly shorter than Tween 80, soluble starch, dodecyl sulphate
Sedimentation time of the sodium to bacterium powder sample.Therefore, polyethylene glycol can be selected as preferred wetting agent.
The determination of protective agent type, by bacterium powder living bacteria count under 50 DEG C of hot conditions of measurement as evaluation index.Tool
Body measuring method are as follows: zymocyte liquid centrifugation gained thallus and protective agent are puddled with the ratio of 1:1 (mass ratio, m/m), are placed in 50 DEG C
In incubator, its living bacteria count is measured at the 1st, 3,5h and the 1st, 3,5d respectively, according to living bacteria count quantity by more to
Few sequence screening best protection agent.Different protectant selections and each protective agent different adsorption cycles living bacteria count referring to
Table 3.
Living bacteria count of the different protective agents of table 3 in different adsorption cycles
This experiment investigation turf, kaolin, sodium alginate, sodium cellulose glycolate are as protective agent to producing yellowish fiber list
Living bacteria count of the born of the same parents bacterium in different adsorption cycles.From table 3 it can be seen that turf, kaolin, sodium alginate, hydroxymethyl cellulose
Sodium can effectively make Cellumomonas flavigena keep effective status, wherein kaolin is at the 1st, 3,5h and the 1st, 3,5d
The distribution of Cellumomonas flavigena living bacteria count amount is respectively (2.23-2.25) × 108A/g, (2.19-2.21) × 108A/g,
More than turf, sodium alginate, sodium cellulose glycolate as protective agent corresponding period living bacteria count.Therefore, it can be selected
Kaolin is as preferred protective agent.
The determination of selected adsorptive support, wetting agent and protective agent dosage, using L9(34) orthogonal experiment, with bacterium powder
Living bacteria count is as evaluation index.On the basis of above-mentioned single factor exploration, with dextrin (A), polyethylene glycol (B), kaolin
(C), temperature (D) is main investigation object, and table 4 gives L9(34) factor of orthogonal test and horizontal.
4 L of table9(34) orthogonal test factor and horizontal
Thallus involved in table 4 and adsorptive support, wetting agent and protective agent dosage are mixed with the ratio of mass ratio (m/m)
It mixes.According to the factor level of table 4, to prepare the living bacteria count of bacterium powder as evaluation index, using L9(34) orthogonal experiment is true
Determine optimal proportion relationship.Table 5 gives L9(34) orthogonal experiments and intuitively analyze result.
5 L of table9(34) orthogonal test and intuitively analyze result
The intuitive analysis K value and very poor R value of table 5 show that carrier dextrin is main investigation factor, and bacterium powder, which is matched, when to be stored
The optimum condition of temperature is A3B1C1D2, separately consider the K of dextrin (A)2With K3The K of value and polyethylene glycol (B)1With K2Value difference is anisotropic
Not significant, i.e. the optimal proportion condition of selection factor A, B, C is determined as 1:1.2,1:0.8,1:0.5.
Embodiment 2
The present embodiment provides a kind of preparation methods of the microbial germ powder of degradable gardens cellulose, comprising steps of 1) bacterium
Liquid fermentation, carries out liquid fermentation Cellumomonas flavigena, stops the viable bacteria of Cellumomonas flavigena in zymocyte liquid when fermentation
Number is not less than 2.0 × 108A/mL；2) thallus separates, and removes the moisture in zymocyte liquid, obtains Cellumomonas flavigena thallus；
3) prepared by bacterium powder, in mass ratio addition adsorptive support, wetting agent and protective agent, stirs and evenly mixs, is dried in vacuo up to described micro-
Biological bacteria powder.
1) bacterium solution is fermented
Reach 2.0 × 10 with living bacteria count8Cellumomonas flavigena (the Cellulomonas of a/mL or more
Flavigena cultivation and fermentation liquid) is seeded in liquid fermentation fermentor as seed liquor with 10% inoculation volume, is led to
It crosses industrial condition of culture to carry out liquid fermentation Cellumomonas flavigena, when bacterium solution living bacteria count reaches 2.0 × 108A/mL
Stop fermentation when above.
It with the determination of carbon source, nitrogen source in culture medium prescription is using cellulase-producing vigor as index screening about liquid fermentation
Gained.Using single-factor variable test method(s), using alternative carbon and nitrogen sources as the factor of changeability combine different carbon and nitrogen sources combinations (because
6) prime information is shown in Table, other culture medium prescriptions and fermentation condition are constant, and by DNS colorimetric method for determining, its corresponding fermentation liquid produces fiber
Plain enzyme activity, the highest carbon and nitrogen sources combination of sequence screening producing enzyme vigor.Measurement result is shown in Table 7.
The different carbon and nitrogen sources factor information of table 6
The different carbon and nitrogen sources combinations of table 7 and corresponding fermentation liquid cellulase-producing vitality test result
In conjunction with table 6 and table 7 it can be concluded that, using the alternative special culture media of carbon and nitrogen sources to Cellumomonas flavigena into
Row liquid fermentation, liquid fermentation culture medium carbon source, nitrogen source select sodium carboxymethylcellulose respectively and ammonium nitrate is best carbon, nitrogen
Source combination, and the feed ratio (mass ratio) of sodium carboxymethylcellulose (CMC-Na) and ammonium nitrate is 5:1.
Based on above-mentioned test result, the present embodiment provides a kind of preferred liquid fermentation medium formula: CMC-Na 5g,
NH4NO3 1g、KCl 0.5g、MgSO4·7H2O 0.5g、KH2PO40.9g, yeast extract 0.5g, distilled water are settled to
1000mL, pH 7.0-7.2.This culture medium is suitable for seed fermentation and scale fermentation.
The condition of culture of seed liquor used in liquid fermentation described in the present embodiment: the fluid nutrient medium of preparation is sub-packed in
In 500mL triangular flask, per bottled 200mL, (121 DEG C, 103MPa, 30min) are handled through moist heat sterilization, it is every under aseptic technique
Bottle inoculation bacteria cake (diameter 1.5cm) 2,30 DEG C of constant temperature, oscillation revolving speed 160rpm, persistently cultivates 36-48h, until seed liquor is effective
Viable count reaches 2.0 × 108Stop fermentation when a/mL or more.
Liquid fermentation culture conditions described in the present embodiment: preparing above-mentioned fermentation culture medium, be placed in automatic fermenter,
Sterilization treatment (121 DEG C, 103MPa, 30min) are carried out according to routine operation, are aseptically accessed with 10% volume fraction
Seed liquor, 30 ± 1 DEG C of constant temperature, mixing speed 250-300rpm, ventilation quantity 0.25-0.30M3/ min, tank press 0.035-
0.055MPa, continuing fermentation 36-48h carry out living bacteria count measurement by blood counting chamber, when bacterium solution living bacteria count reaches
2.0×108Stop fermentation when a/mL or more.
2) thallus separates
Above-mentioned gained zymocyte liquid is stored at room temperature, and uses high speed frozen type butterfly seperator, is turned with 3500rpm
Speed, 5 ± 1 DEG C of centrifuging temperatures carry out high speed centrifugation 10min, remove the moisture in fermentation liquid, obtain lark paste thallus.
In this step, described to be stored at room temperature, 60-100min is stood in butterfly seperator under the conditions of referring to 20-25 DEG C.
The purpose being stored at room temperature is to realize efficiently separating for thallus and moisture in fermentation liquid, and avoids the damage of thallus to the greatest extent
It loses.The paste thallus obtains the bacterium mud for containing about 10% moisture after referring to low-temperature and high-speed centrifugation.In thallus separation process, closely
The aqueous solution on bacterium mud upper layer is the bacterium solution of high concentration, causes thallus to lose if being completely separated.In addition, bacterium powder preparation test card
Bright, the bacterium mud containing about 10% moisture and adsorptive support, protectant absorption adhesiveness are preferable, and have hydrophily, easily with
Wetting agent polymerization.Therefore, in the present embodiment thallus separating step, gained thallus generally refers to the bacterium mud containing about 10% moisture.
3) prepared by bacterium powder
Adsorptive support, wetting agent and protective agent is added in above-mentioned gained thallus in mass ratio, by quality ratio, the bacterium
The ratio of body and adsorptive support is 1:(0.5-1.5), the ratio of the thallus and wetting agent is 1:(0.5-1.5), the bacterium
Body and protectant ratio are 1:(0.5-1.5).It stirs and evenly mixs, is dried in vacuo, obtain off-white color microbial germ powder.In this step
In, vacuum drying parameter is preferred are as follows: under the conditions of -0.08MPa, 35 DEG C of dry 24-36h.
Embodiment 3
The present embodiment provides the evaluation method of the microbial germ powder of Examples 1 and 2 preparation.The present embodiment is weightless using degradation
Rate method evaluates microbial germ powder to the degradation efficiency of afforestation waste, by different Storage periods (30d, 90d, 150d, 210d,
270d, 330d, 390d and 450d) microbial germ powder be each configured to 5 ‰ bacterium powder suspensions, respectively at constant temperature (30 ± 1 DEG C)
Its degradation weight-loss ratio to afforestation waste, its drop of the weightless bigger explanation of ratio are calculated when 5d, 10d and 15d of culture
It is higher to solve efficiency.
Afforestation waste described in the present embodiment refers to reed (Phragmites australis), tulip
(Tulipa gesneriana), silver wire grass (Chloranthus japonicus), iris (Iris tectorum) one kind or
Several withered stalks.Pollution and interference in order to avoid other miscellaneous bacterias to selected materials are implemented in degradation efficiency evaluation procedure
The preceding withered stalk to selection does sterilization treatment, and is dried in vacuo.Preferably, Biocidal treatment method is moist hear heat test, sterilizing
Condition is 121 DEG C, 103MPa, 30min.Vacuum drying set temperature is 75 DEG C, and vacuum degree is -0.08MPa, and drying time is
24~36h.
In order to further clarify the present embodiment the method, make its more operability and practicability.Involved by the present embodiment
Chemical reagent be analysis it is pure；Water used in compounding medicine is deionized water, and the water as used in phosphate buffer is
Deionized water；It is sterile water that sterile water, which impregnates water used in withered stalk, i.e. the deionized water of moist hear heat test processing.
Specifically, afforestation waste degradation weight-loss ratio measuring method is as follows: the sterile water logging of afforestation waste
Bubble overnight, is dried under vacuum to constant weight, shreds to the section for being about 2cm；It is sub-packed in above-mentioned in 500mL triangular flask to degradable material,
Each bottled 5g of triangle is above-mentioned to degradable material；(NH is added into each triangular flask4)2SO4 0.2g、MgSO4·7H2O
Phosphate buffer (PBS) 40mL of 0.05g, 5mmol/L pH=7.0.Every bottle is inoculated with 5 ‰ bacterium powder suspension 10mL respectively, to connect
Kind 10mL sterile water is placed on 30 ± 1 DEG C of constant temperature incubations as blank control group (CK), mixing, and routine observation simultaneously measures withered grass
Form and quality change situation.Every processing test is repeated 3 times.The evaluation result of the present embodiment is shown in Table 8.
Degradation weight-loss ratio calculation formula: D=(M0-M1)/M0 × 100%.Wherein, D indicates afforestation after periodically culture
The degradation weight-loss ratio of waste, %；M0 indicates sterile water soaked overnight, and filtering, afforestation is useless before vacuum drying gained is handled
Gurry quality, g；After M1 indicates periodically (5d, 10d and 15d) culture, sterile water washes away thallus and spore, filters, vacuum drying
The undegraded afforestation waste quality of gained, g.
The different Storage period microbial germ powders of table 8 evaluate the degradation effect of afforestation waste
Table 8 shows, to 30d, 90d, 150d, 210d, 270d, 330d, 390d under the conditions of being stored in room temperature, being protected from light and
The microbial germ powder of 450d can effectively degrade afforestation waste.Degradation energy of the microbial germ powder to afforestation waste
Power is reduced with extending with for Storage period, degradation of the microbial germ powder to afforestation waste in 390d Storage period
It is close to the degradation weight-loss ratio of afforestation waste that weight-loss ratio is higher than the microbial germ powder in 30%, 450d Storage period
30%.In same Storage period, degradation weight-loss ratio prolonging with incubation time of the microbial germ powder to afforestation waste
It grows and increases, cultivate effective degradation of achievable afforestation waste after 15d.
Further narration is done to the present invention above in conjunction with embodiment, but present invention is not limited to the embodiments described above,
Within the knowledge of one of ordinary skill in the art, it can also make without departing from the purpose of the present invention
Various change.
Claims (5)
1. a kind of microbial germ powder evaluation method for promoting garden waste degradation-type, which is characterized in that afforestation waste
Microbe inoculation bacterium powder is placed in 30 ± 1 DEG C of constant temperature incubations, the mass change of results of regular determination afforestation waste degradation residue
Situation, by degrading, weight-loss ratio evaluates the microbial germ powder to the degradation efficiency of afforestation waste；
When cultivating 15d, the microbial germ powder is not less than 30% to the degradation weight-loss ratio of afforestation waste；
The microbial germ powder the preparation method comprises the following steps: by Cellumomonas flavigena (Cellulomonas flavigena) thallus with
Adsorptive support is mixed with the ratio of quality 1:(0.5-1.5)；Add wetting agent, the ratio of the thallus and wetting agent is 1:
(0.5-1.5)；Protective agent is added, the thallus and protectant ratio are 1:(0.5-1.5)；It stirs and evenly mixs, is dried in vacuo
Up to the microbial germ powder；
The adsorptive support selects dextrin, diatomite, talcum powder or calcium carbonate；
The wetting agent selects Tween 80, polyethylene glycol, soluble starch or lauryl sodium sulfate；
The protective agent selects turf, kaolin, sodium alginate or sodium cellulose glycolate.
2. promoting the microbial germ powder evaluation method of garden waste degradation-type according to claim 1, which is characterized in that be used in
10-30 DEG C is protected from light the microbial germ powder degradation afforestation waste for storing 390d, and the microbial germ powder is green to gardens
The degradation weight-loss ratio for changing waste is not less than 30%.
3. the microbial germ powder evaluation method according to claim 1 or claim 2 for promoting garden waste degradation-type, which is characterized in that
The afforestation waste is reed (Phragmites australis), tulip (Tulipa gesneriana), silver wire
One or more of withered stalks of careless (Chloranthus japonicus), iris (Iris tectorum).
4. application of the microbial germ powder described in claim 1 in degradation afforestation waste.
5. application according to claim 4, which is characterized in that the microbial germ powder is with Cellumomonas flavigena
(Cellulomonas flavigena) thallus is as active bacteria, for the cellulose in afforestation waste of degrading.
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