CN105112311A - Pichia yeast YPD-YL2 and application of same in biological deodorization - Google Patents
Pichia yeast YPD-YL2 and application of same in biological deodorization Download PDFInfo
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- 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
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
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- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/59—Biological synthesis; Biological purification
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- 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
- Y02W30/20—Waste processing or separation
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- 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 provides a Pichia yeast YPD-YL2 and application of the same in biological deodorization. The Pichia yeast YPD-YL2 has a Latin name of Pichia sp. and an accession number of CCTCC M 2015225. The Pichia yeast YPD-YL2 provided by the invention is separated from an environment of a refuse landfill, is capable of removing foul gas produced by garbage decomposition and can decompose garbage and produce slightly smelly gas. Repeated addition of the Pichia yeast YPD-YL2 is not needed in deodorization with the Pichia yeast YPD-YL2, so workload is mitigated. Due to security of the Pichia yeast YPD-YL2, the Pichia yeast YPD-YL2 can also be used for treating substances like animal feeds and additives so as to allow the substances to be edible to animals.
Description
Technical field
The present invention relates to field of deodorization, especially, relate to a kind of pichia spp YPD-YL2 and the application in biological deodorizing thereof.
Background technology
Odor pollution is one of universally acknowledged seven overall situation public hazards, and public lavatory, livestock and poultry farm, refuse landfill and refuse collection station often give out stench niff, become the main place of odor pollution.Containing multiple toxic gas in foul gas, wherein main component is ammonia and hydrogen sulfide, causes great impact to the health of the resident of periphery, citizen and sanitationman.Effective process odor pollution, reduces the impact protection people healthy of odor pollution on ecotope and has become a social concern urgently solved.
At present for the solution of odor pollution problem, conventional method has traditional physico-chemical process and the large class of biological deodorizing two.Typical physical chemistry deodorizing method antibacterial solid deodorizer disclosed in CN20091173160.5, generation due to foul gas produces when mostly being all kinds of refuse of microbial decomposition, thus the application of this China provides a kind of composition containing tensio-active agent, in said composition, tensio-active agent used can realize absorption to foul gas molecule, can play antibacterial effect again to the microorganism producing foul gas.Thus realize the concentration reducing foul gas.But this solid deodorizer in use, can there is saturated problem in adsorption effect, and along with the prolongation of duration of service, antibacterial effect also can weaken along with the reduction of effective constituent concentration.Effect is difficult to lastingly.The deodorizing method also had is for carrying out oxidative degradation to stench composition by introducing external source Strong oxdiative thing or aromatoising substance or cover, reach deodorizing effect, by force pointed, the feature of instant effect, but it is little also to have range of application, easily cause the shortcoming of secondary pollution.
Biological deodorizing principle be the biological enzyme that produces of function bacterial classification by filtering out from stench environment or bacterial classification to niffy gas or material directly degrade, adsorb or stench is removed to the suppression of producing stench microorganism.Relative to the physico-chemical process of conventional process stench, microorganism constantly breeds, and biological deodorizing effect is comparatively lasting, non-secondary pollution, and easy to use, with low cost, becomes the study hotspot that stench controls.Report about biological odor removal method sees nineteen fifty-seven American scientist the earliest and utilizes soil microorganisms process H
2s waste gas, and patented.Through the development of decades, different kind organism solid deodorizer has been applied to many aspects such as production and life, and achieves obvious effect.Such as, provide strain deamination deodorizing bacterial strain QDN01 and the application in biological deodorizing thereof in CN201410344493.0, this bacterium has stronger Decomposition for containing ammoniacal substance, also to the stink substance that the ight soil of poultry produces, there is stronger Decomposition simultaneously, find that this bacterial strain can decompose the amine substance in starch-containing chicken manure preferably through experiment, reduce stink.But this bacterium is only when containing nutritive substances such as starch, just can reaches optimum discomposing effect, add use cost.
Summary of the invention
An object of the present invention is to provide a kind of pichia spp removing nitrogen and hydrogen sulfide.
An object of the present invention is to provide the application of pichia spp YPD-YL2 in biological deodorizing.
An object of the present invention is to provide a kind of solid deodorizer containing this thalline.
An object of the present invention is to provide a kind of composite fungus agent containing this bacterium.
An object of the present invention is to provide the application of this composite fungus agent a kind of in deodorizing.
According to an aspect of the present invention, provide a kind of pichia spp, pichia spp is pichia spp YPD-YL2, and the Latin of pichia spp is called Pichiasp., and preserving number is CCTCCM2015225.
Additionally provide the application of above-mentioned pichia spp in biological deodorizing according to a further aspect in the invention.
Further, biological deodorizing comprises the deodorizing process carried out feed, rubbish or organic fertilizer.
Additionally provide a kind of solid deodorizer according to a further aspect in the invention, the bacterium liquid containing above-mentioned Pichi strain is adsorbed on carrier and obtains.
Further, be 1: 1 ~ 20 to mix by the volume of bacterium liquid and the mass ratio of carrier.
Additionally provide a kind of composite fungus agent according to a further aspect in the invention, comprise the fermented liquid of above-mentioned pichia spp YPD-YL2, lactobacillus and subtilis.
Further, pichia spp YPD-YL2 fermented liquid, lactobacillus and subtilis are 2 ~ 7: 1 ~ 3: 1 mixing by biomass ratio.
Further, pichia spp YPD-YL2 fermented liquid, lactobacillus and subtilis are 3: 2: 1 mixing by biomass ratio.
Further, in composite fungus agent liquid fermentation medium used, at least comprise corn stalk powder 50 ~ 150g/L, wheat bran 20 ~ 70g/L, brown sugar 80 ~ 120g/L, tomato powder 40 ~ 60g/L.
Additionally provide the application of a kind of above-mentioned composite fungus agent in deodorizing according to a further aspect in the invention.
The present invention has following beneficial effect:
Pichia spp YPD-YL2 provided by the invention fills up the environment of field to be separated from rubbish to obtain, the foul gas that this bacterium produces for garbage decomposition has good removal effect on the one hand, this thalline itself can also decompose rubbish simultaneously, produce the less gas of stink simultaneously, when using this thalline to carry out deodorizing, without the need to repeatedly adding, alleviate workload.Simultaneously due to the security of this bacterium, for the treatment of the such as material such as animal-feed, additive, animal edible can also be more suitable for make it.
Except object described above, feature and advantage, the present invention also has other object, feature and advantage.Below with reference to experimentation, the present invention is further detailed explanation.
In the present invention, pichia spp used (Pichiasp.) YPD-YL2 bacterial strain is kept at China typical culture collection center (abbreviation on April 12nd, 2015, CCTCC), depositary institution address: Hongshan District, Wuhan City, Hubei Province Bayi Road Wuhan University China typical culture collection center, postcode 430072.Deposit number is the Classification And Nomenclature of this bacterium of CCTCCM2015225 is pichia spp (Pichiasp.).
Embodiment
Below in conjunction with experimentation, embodiments of the invention are described in detail, but the multitude of different ways that the present invention can be defined by the claims and cover is implemented.
If do not specialize, the conventional means that technique means used in embodiment is well known to those skilled in the art.
The percentage sign " % " related to herein, if not specified, refers to mass percent; But the per-cent of solution, unless otherwise specified, refers in solution 100ml containing solute some grams; Per-cent between liquid, refers to the ratio of capacity 20 DEG C time.
This bacterial strain is obtain from screening and separating the percolate deriving from the refuse landfill of Changsha Mark Reserved.Screening used medium is YPD substratum (YeastExtractPeptoneDextroseMedium, yeast extract powder peptone dextrose culture-medium).After gained thalline is bred on YPD substratum; gained bacterium colony outward appearance is that oyster white, bacterium colony are large and thick; local protuberance local, surface is smooth; moistening; through Environmental Biotechnology institute of Changsha Environmental Protection College, Molecular Identification is pichia spp; be numbered: YPD-YL2; utilize pcr amplification fungi Internal Transcribed Spacer (the transcribed spacer ITS1 between 28S and 18S gene); its upstream primer is: TCCGTAGGTGAACCTGCGG, and downstream primer is: TCCTCCGCTTATTGATATGC.Gel electrophoresis after amplification, reclaim product band, genome company is sent to check order, its sequencing result carries out sequence alignment in NCB1 gene library, find DNA sequence dna and its homology of only having a strain Pichi strain, homology is 77%, and does not retrieve DNA sequence dna and its homology of other microorganism.Illustrate that this bacterial strain is with pichia spp homology and for newfound bacterial strain.Gained gene order lists in gene order matrix section.The DNA sequence dna of this bacterial strain is DNA sequence dna: CATTGGCATCTACTGATCTGAGTCGAGCTCATATGGCTCCGAACCCCAAGCGTCCT GGTCCAATTCCCTCCGGGCCTCCTTTCCTTTCCGCGGGGCCGTGGCCCGGCCCACT CTGCCCAACTCCCCTCCTGGCAGAAAGAAACCACCCTCCAAACGGCCTGGCCCCCC GAATGGCCAACGGCGCCATGGGCGTTCCAGAACTCCATGAATCCCCATGGCTGGCA TTCCCACTAAGTATCCCATTTCCCTGCCCTCCTCCTCCATGCCAAAACCCAAAAAT CCCTTGGTGAAAGTTTTGGGGTAAAATAAAAACTCCTGAACTAATATACCTGGTTG GGTGGTGGGGGCCCTCCCCCCATGGGGAACAAAAAACCCACTAAAGGTCCCTTCCC AAGGTTCCCCACAGAAAAG.
Following substratum uses as required.Subsequent experimental is the deodorizing effect illustrating that pichia spp YPD-YL2 has, and carries out containing the solid deodorizer of pichia spp YPD-YL2, the operation strategies of composite fungus agent and effect thereof, and real protection scope is not limited to listed technical scheme in experiment.
The substratum of this bacterial strain:
YPD liquid nutrient medium: in the substratum in 1000ml, containing yeast extract powder 5g, peptone 10g, glucose 20g, adjust ph is about 5.4, room temperature preservation after sterilizing, obtains YPD liquid nutrient medium;
YPD solid medium: in the substratum in 1000ml, containing yeast extract powder 5g, peptone 10g, glucose 20g, after adjust ph is about 5.4,14g agar powder, after sterilizing, Bechtop is down flat plate, and rear enclosed is preserved.
YPD-YL2 strain culturing:
Strains separation is cultivated: with transfering loop or inoculating needle streak inoculation in containing in YPD solid medium flat board, culture temperature is 30 DEG C, and incubation time is 2-3 days, obtains mono-clonal bacterium colony;
Inoculation is cultivated: picking mono-clonal bacterium colony, is inoculated in the sterile test tube containing 5ml, 30 DEG C, 200rpm shaking culture 48 ~ 72 hours, muddy to substratum;
Bacterium liquid is preserved: by above-mentioned containing the liquid nutrient medium of bacterial strain and the glycerine of sterilizing in 7: 3 ratio mix in Bechtop after, be kept in-80 DEG C of Ultralow Temperature Freezers;
Fermented liquid is produced: bacterial classification, after spreading cultivation, is inoculated in 50L fermentor tank by 1%, aerobic fermentation, cultivates about 3 days for 30 DEG C.
Zymocyte liquid used herein: with transfering loop or inoculating needle streak inoculation in containing in YPD solid medium flat board, culture temperature is 30 DEG C, incubation time is 3 days, obtain mono-clonal bacterium colony, picking mono-clonal bacterium colony, is inoculated in the sterile test tube containing 5ml, 30 DEG C, 200rpm shaking culture 48 ~ 72 hours, muddy to substratum.Bacterial classification, after spreading cultivation, is inoculated in 50L fermentor tank by 1%, aerobic fermentation, cultivates about 3 days, obtains pichia spp YPD-YL2 fermented liquid for 30 DEG C.If can be undertaken by the production method of a small amount of deodorant liquid when required fermentation liquid measure is less: by bacterial classification after slant medium activation, be inoculated in the triangular flask that 500ml liquid culture medium is housed respectively, be placed in constant-temperature table incubator, under 30 DEG C of constant temperatures, within 200r/ minute, cultivate and both obtained pichia spp YPD-YL2 fermented liquid in 3-5 days.Bacterial classification increasing expansion culturing step can carry out amplification cultivation with reference to method disclosed in CN201210030324.0 and obtain corresponding and fermented liquid.
Cadmium hydroxide absorption liquid herein: weigh 4.3g Cadmium Sulphate (3CdSO
48H
2o), 0.3g sodium hydroxide and 10g ammonium alcohol polyvinyl phosphate be soluble in water respectively, faces the used time, and by three kinds of molten liquid-phase mixing, strong jolting to complete miscibility, then is diluted with water to 1L and takes.This solution is white suspension, and each used time wants strong jolting evenly to measure again, stores in refrigerator and can preserve one week.
Sensory evaluation method used is for convening 100 osmatic volunteers herein, carries out human body sense of smell identification respectively to different treatment sample.Statistics averaged after impression is marked.Wherein standards of grading are: 5 points represent ordorless; 4 points represent slight stench; There is stench in 3 points of representatives; 2 points of representatives do not have the stench of change; 1 point represents stench and strengthens.Statistics is divided into three grades: +++ represent strong foul smell, obatained score is 1 ~ 2 point; ++ represent foul smell, obatained score is 3 ~ 4 points; + representing slight foul smell, obatained score is 4 ~ 5 points.
Experiment 1, pichia spp YPD-YL2 fermented liquid are to ammoniacal liquor and hydrogen sulfide degradation effect
Get 18ml bacterium liquid in 2L large beaker, and add the ammoniacal liquor 2ml that concentration is 1000ml/L, then put in large beaker small beaker that 20ml concentration is the sulfuric acid absorption liquid of 0.005mol/L (sulfuric acid absorption liquid: measuring 2.8ml density is that the vitriol oil of 1.84g/ml is added to the water dilution and is settled to 1L is housed.Face used time redilution 10 times), cover double-plastic by large beaker sealing sealing, carry out falling ammonia experiment.
Get 18ml bacterium liquid and put into 2L large beaker, and add the hydrogen sulfide solution that 2ml concentration is 100mg/L, in large beaker, put into the 50ml small beaker that 20ml cadmium hydroxide absorption liquid is housed afterwards again, cover double-plastic sealing, carry out and fall hydrogen sulfide production test.
When carrying out two experiments above, all using the sterilized water of equivalent, alternative cadmium hydroxide absorption liquid and sulfuric acid absorption liquid, as blank group, are tested under the same conditions respectively.Experiment condition is: the incubator being placed in 30 DEG C is cultivated, each process repetition 3 times.After 24 hours, take out the small beaker in large beaker respectively, adopt reagent colorimetric method to measure the clearance of ammonia, adopt methylene-blue colorimetric method to measure the clearance of hydrogen sulfide, acquired results is listed in table 1.
Table 1 deodorant liquid ammonia and hydrogen sulfide removal rate result table
| Detect number of times | 1 | 2 | 3 | Blank group | On average |
| Ammonia removal rate (%) | 96.1 | 95.5 | 95.3 | 0 | 95.6 |
| Hydrogen sulfide removal rate (%) | 72.5 | 72.1 | 73.1 | 0 | 72.6 |
Therefore use pichia spp YPD-YL2 provided by the invention can ammonia effectively in decomposition gas and hydrogen sulfide, the clearance on average for ammonia can reach 95.6%, can reach 72.6% to the clearance of hydrogen sulfide.Relatively do not use the removal effect of blank sample two kinds of gases of fermented liquid obvious.This may be higher with the institute's utilization ratio of mycetome to ammonia and hydrogen sulfide in fermented liquid, thus improve the removal efficiency to Ammonia in Air gas and hydrogen sulfide.In experimentation, beaker used is all in sealed state, and this is conducive to the loss preventing ammonia and hydrogen sulfide, is conducive to the interference preventing other gases to fermented liquid decomposition result.Experimental results is reliable.
Experiment 2, pichia spp YPD-YL2 are to the deodorizing effect of domestic refuse
The deodorizing effect of domestic refuse detects: domestic refuse derives from the food waste of school lunch service, and main component is the materials such as leftover, fruits and vegetables scurf, plastics bag and wood fibre.Be be mixed to get deodorizing diluent at 1: 3 by pichia spp YPD-YL2 fermented liquid and water volume ratio, get the deodorizing diluent of Fresh Life rubbish total mass 10%, it is evenly sprayed in domestic refuse, realize inoculation.Simultaneously with the same terms, to dilute deodorant liquid water used in contrast, under same experimental conditions, carry out blank assay.Respectively at latter 4,8,24 hours of inoculation, sensory evaluation method is adopted to measure foul smell grade.The results are shown in Table 2.
Table 2 pichia spp YPD-YL2 fermented liquid is to domestic refuse deodorizing result table
| Handling duration (hour) | 0 | 4 | 8 | 24 |
| Pichia spp YPD-YL2 fermented liquid | +++ | ++ | ++ | + |
| Water | +++ | +++ | +++ | +++ |
Note: +++ represent strong foul smell; ++ represent foul smell; + represent slight stench taste
From table 2, before process, domestic refuse stench is strong, employ the domestic refuse after pichia spp YPD-YL2 fermentation liquor treatment provided by the invention, along with the increase of handling duration, the odor strength that rubbish produces reduces, gradually when inoculation is after 24 hours, the stink of rubbish is down to slightly, substantially reaches human body tolerance interval.It can thus be appreciated that, in the fermented liquid of pichia spp YPD-YL2 provided by the invention, institute's mycetome itself can make full use of on the one hand each type organic in rubbish as Carbon and nitrogen sources needed for self, carry out decomposition to rubbish to utilize, the enzyme classes that thalline produces in process of growth on the other hand can also be decomposed the material that thalline can not directly utilize, be beneficial to follow-up thalline utilize, thus when without the need to adding any extra carbon and nitrogen sources, improve pichia spp YPD-YL2 fermented liquid to the capacity of decomposition of rubbish.Realize the decomposition to the material producing foul gas in rubbish.On the other hand, pichia spp YPD-YL2 enzyme classes that thalline produces, can also decompose the main component in the foul gas produced, thus reduces the concentration of the foul gas produced.Two kinds of effect synergies, improve the sustainability of the effect that garbage odor after treatment reduces, and without the need to repeatedly adding fermented liquid, new stench can also be suppressed to be formed.
Experiment 3, pichia spp YPD-YL2 fermented liquid obtain the desulfurization of solid deodorizer
Pichia spp YPD-YL2 fermented liquid obtains the making of solid deodorizer: after getting pichia spp YPD-YL2 fermented liquid 100ml and 200g wood chip Homogeneous phase mixing, at 35 DEG C, drying is carried out to gained wood chip, when being 60% to its water ratio, obtain solid deodorizer, in follow-up desulfurization.Simultaneously obtain blank wood chip, as blank experiment blank deodorizing microorganism used with aqua sterilisa according to as above method drying.Live body bacterium amount in gained solid deodorizer can reach 400,000,000/more than g.
Solid deodorizer desulfurization: by above-mentioned solid deodorizer and blank deodorizing microorganism, evenly be laid in 2 2L large beakers respectively, and add the hydrogen sulfide solution 2ml that concentration is 100mg/L, the 50ml small beaker that 20ml cadmium hydroxide absorption liquid is housed is put into again in large beaker, cover double-plastic sealing, carry out and fall hydrogen sulfide production test.The incubator being placed in 30 DEG C is cultivated.Each process repetition 3 times.After 24 hours, take out small beaker, adopt the clearance of methylene radical prison colorimetric method for determining hydrogen sulfide.The results are shown in Table 3.
Table 3 solid deodorizer is to the clearance result table of hydrogen sulfide
| Test item | 1 | 2 | 3 | Blank | On average |
| Hydrogen sulfide stripping rate (%) | 56.2 | 55.8 | 57.3 | 0 | 56.4 |
From table 3, after solid deodorizer made by pichia spp YPD-YL2 fermented liquid provided by the invention, although decrease for the relative fermented liquid of removal efficiency of hydrogen sulfide itself, but still higher than blank experimental result.It can thus be appreciated that, after pichia spp YPD-YL2 fermented liquid provided by the invention can make solid deodorizer as stated above, for the removal to foul smell in the environment being unfavorable for this growth.The biological desulphurization deodorizing of such as ight soil or biological deodorizing filter tank, bio-trickling filter.In table 3, hydrogen sulfide stripping rate is lower than the result in table 1, and this may be because institute's mycetome quantity in thalline fermented liquid is higher than the thalline content in solid deodorizer obtained after treatment.In the deodorizing of obvious gained solid, used carrier can be not limited to wood chip, can also be that other biological acceptable carrier is as wilkinite, diatomite etc.The blending ratio of carrier and bacterium liquid can for bacterium liquid and described carrier by volume mass ratio be 1: 1 ~ 20 to mix.
Preferably, when solid deodorizer is used for domestic refuse or night soil-treatment, can according to 1 ~ 2% (V/V) of pending rubbish or ight soil, by even for solid deodorizer spreading to rubbish or ight soil surface.This solid deodorizer be used for biological deodorizing filter tank, bio-trickling filter biological desulphurization deodorizing time can the add-on of sweetening agent routinely add.
Experiment 4, composite fungus agent are used for rubbish Deodorization Experiment
One aspect of the present invention additionally provides a kind of composite fungus agent containing above-mentioned pichia spp YPD-YL2.It is 2 ~ 7: 1 ~ 3: the 1 pichia spp YPD-YL2 fermented liquid mixed, lactobacillus and subtilis that this microbial inoculum at least comprises by biomass ratio.Obviously this composite fungus agent may be used in other deodorization composite bacteria agent, as the composite fungus agent obtained to this cooperation being used for disclosing in CN201310293372.3 in biological deodorizing microbial inoculum.Be more preferably 3: 2: 1.Below be mixed to get composite fungus agent by biomass than 3: 2: 1 to test.
Composite fungus agent used medium: at least comprise corn stalk powder 50 ~ 150g/L, wheat bran 20 ~ 70g/L in this composite fungus agent liquid nutrient medium used, brown sugar 80 ~ 120g/L, tomato powder 40 ~ 60g/L.
To contain corn stalk powder 100g, wheat bran 50g in 1000ml substratum, brown sugar 100g, tomato powder 50g are formula, and dilute with water obtains liquid nutrient medium.The substratum of this formula can also be used in the process of this composite fungus agent of suitability for industrialized production, and effect is more excellent.
The production of composite fungus agent: by the composite fungus agent after spreading cultivation, being inoculated in the aforesaid liquid substratum of 50L by kind of amount by 1wt.%, utilizes this liquid nutrient medium at 30 DEG C, carry out aerobic fermentation and cultivates 3 ~ 5 days.
The deodorizing of domestic refuse: domestic refuse used and laboratory facilities identical with experiment 2.Carry out subjective appreciation.Acquired results is listed in table 4.
Table 4 composite fungus agent is used for domestic refuse deodorizing sensory evaluation scores result table
| Handling duration (hour) | 0 | 4 | 8 | 24 |
| Composite fungus agent | 1 | 4 | 4 | 5 |
| Blank test (water) | 1 | 1 | 1 | 1 |
Note: +++ represent strong foul smell; ++ represent foul smell; + represent slight stench taste
From table 4, composite fungus agent provided by the invention has good deodorizing effect for domestic refuse, can along with the increase of handling duration, and the stink of handled domestic refuse reduces gradually.And owing to employing composite fungus agent, the deodorizing effect of rubbish is owing to testing the result in 2.After treated 24 hours, stench, for reaching close to not having, can also produce the fragrance of similar wine simultaneously.Original residual odors taste is carried out comprehensively, thus reaches 5 grades of ordorless appraisal result.After being coordinated by above-mentioned formula with bacterial classification provided by the invention by multiple bacterium, the living environment of pichia spp YPD-YL2 bacterium can be improved on the one hand, it is made better to discharge enzyme repugnant substance to Decomposition, on the other hand can lactobacillus and subtilis itself also can under pichia spp YPD-YL2 produces the inducing action of enzyme, produce the enzyme to decomposing further through pichia spp YPD-YL2 decomposition after product, form aroma, carry out comprehensively, finally reaching the effect of odorless with residual stench.
Test 5, utilize composite fungus agent to produce fertilizer experiment to livestock manure fermented
Experimental technique: the method empirically in 5 obtains composite fungus agent, be be mixed to get dilution composite fungus agent at 1: 3 by volume by composite fungus agent and water, compound is obtained afterwards by after feces of livestock and poultry (as chicken manure) and straw powder (dry weight 1: 3) mixing, be 1: 10 mixing by compound and dilution composite fungus agent volume ratio, the backsight humidity increase and decrease water yield or straw powder amount, maintaining fermentation system humidity is 60%.Carry out compost fermentation 3 ~ 7 days, gained fermentation fertilizer acid fragrance weight, the stink of chicken manure alleviates.
Adopt above-mentioned composite fungus agent to ferment to feces of livestock and poultry, the stink in ight soil can be alleviated on the one hand, improve the acceptability of operator when using.Find through experiment, after gained fertilizer being used for infertile field, the growing state of plant is identical with other fertilizer growing ways that there is strong foul smell of use simultaneously.Illustrate that gained fertilizer is after removing stink, can't impact the performance of material infertile field effect in wherein contained ammonia.
The preparation of experiment 6, deodorant addtives
The present invention additionally provides a kind of deodorant addtives containing above-mentioned composite fungus agent on the other hand.The fermentation material of concrete composite fungus agent comprises after the impurity elimination respectively of the raw materials such as maize straw, wheat bran, Semen Maydis powder, tomato powder, rice bran and dregs of beans, pulverizing, by the mixing of 20 ~ 40 parts by weight Corn stalks, 5 ~ 15 weight part wheat bran, 20 ~ 35 parts by weight Corn powder, 1 ~ 10 weight part tomato powder, 10 ~ 20 weight part rice brans and 10 ~ 20 weight part dregs of beans, obtain fermentation material.
The preparation of composite fungus agent stoste: getting in experiment 5 by biomass ratio is 3: 2: 1 composite fungus agents be mixed to get, and adds in the brown sugar water of 1wt.%.By composite fungus agent: brown sugar water gross weight ratio is 8: 100, keep 30 DEG C of water temperatures 1 ~ 2 hour, bacterial classification is rejuvenated.Obtain composite fungus agent stoste.
The preparation of deodorant addtives: by composite fungus agent stoste by composite fungus agent stoste and the weight ratio of fermentation material be 4: 1000 mix after, to drop in horizontal single paddle mixer mixing 15 minutes, after mixing, the uniformity coefficient variation coefficient reaches less than 10%.Gained mixture is loaded in ferment at constant temperature bucket, sealing thermal insulation 30 DEG C, send out alcohol and obtain deodorizing biotic additives after 3 days, pack packing.
Gained deodorant addtives can use as feed separately, also can use as other additives beyond animal-feed.According to existing research conditions, can predict, due to containing pichia spp YPD-YL2 provided by the invention in gained deodorant addtives, gained additive smell is more conducive to animal and accepts, pichia spp YPD-YL2 can also play the effect of the yeast that it has simultaneously, improve the intestinal health of animal body, promoting digestion absorbs, and driven interior of articles completes improvement.Thus realize it as other of fodder additives as increased meat or urge long effect.Certainly the preparation method of this additive is not limited to above-mentioned, also the composite fungus agent provided in experiment 5 can be used in the preparation of other animal additives.
Greenfeed experiment is produced in experiment 7, composite bacteria fermentation stalk
The preparation of composite fungus agent stoste: identical with experiment 6, obtains composite fungus agent stoste.
Fermentation raw material prepares: maize straw, sweet potato vine, beans bar and beanpod, Caulis arachidis hypogaeae, green grass, rice straw etc., regulate its water content between 60 ~ 70% by modes such as Exposures to Sunlight, all can be used as fermentation greenfeed raw material.
Fermentation greenfeed: composite fungus agent stoste distilled water diluting 200 is doubly made composite fungus agent diluent, then loads greenfeed raw material, once-combined microbial inoculum diluent is sprayed in every raising about 8 centimetres.Filling buttress is inner, and the add-on amount of composite fungus agent diluent is that composite fungus agent diluent and greenfeed raw materials quality ratio are about 4: 1000, until fill fermenting container, then steady and sure, suitably sprays a little composite fungus agent diluent in the top of charge level more.The ratio of filling buttress top composite fungus agent diluent and raw material is about 8: 1000, covers plastics film, sealing and fermenting about month, has when fragrance or salted vegetables taste just can enable until ensilage fermenting-ripening.
Being only with top fermentation greenfeed process and illustrating, also can be the fermenting process of other greenfeeds existing.Adopt above-mentioned fermenting agent to ferment to all kinds of plant, gained greenfeed fragrant, improve the acceptance level of animal body.Make its appetite better, thus realize the weightening finish of animal body further and increase effect.Simultaneously gained greenfeed also has listed the every of animal additive in experiment 6 and improves effect, thus improves the physique of animal body comprehensively, reaches the effect of diseases prevention.
Additive in concrete this use experiment 6 still tests the greenfeed in 7, can select according to the kind of concrete animal and eating habits thereof.Use and all can reach above-mentioned effect afterwards.
From above-mentioned experiment 1 ~ 7, pichia spp YPD-YL2 mono-aspect provided by the invention has the effect of decomposing hydrogen sulfide and ammonia, the material producing foul gas in rubbish can also be decomposed simultaneously, in the process of decomposing, avoid the generation of foul gas, thus reach the effect for the treatment of both principal and secondary aspect of disease.Pichia spp YPD-YL2 well-grown in the environment containing rubbish in its available fruits and vegetables, can not affect its growth and the discomposing effect to foul gas due to the bad environments of rubbish simultaneously.The present invention simultaneously additionally provides on the other hand and better can adapt to the composite fungus agent that treating refuse produces stench.In this composite fungus agent, contained multiple bacterium and pichia spp YPD-YL2 work in coordination with and play a role, and after using this composite fungus agent, stench removing effect is owing to being used alone pichia spp YPD-YL2.Composite fungus agent is except playing the effect of deodorizing simultaneously, can also be used in the fermentation of feed needed for animal body, by all kinds of peculiar smell removings lower for animal body acceptance, can improve animal body food-intake.Further can also improve the intestinal environment of animal body, reach increasing meat, the effect of growth promoting effects.This composite fungus agent can also be used for the fertilizer that ferments simultaneously, and in gained fertilizer, repugnant substance content reduces, and decreases the stench injury that fertilising personnel are subject in fertilising process.Although gained has niffy substances content in fertilizer to reduce, when this material is for fertilizing the soil, effect can't be had a strong impact on, and substantially can reach the result of use identical with common fertilizer.In above experimentation, concrete preparation method used is and illustrates, other are that the method for illustrating all can be used in corresponding experiment, obtain the product with above-mentioned effect.
Embodiment
In following examples, material used and instrument are commercially available.In following examples, non-detailed portion can consult the relevant discussion of elsewhere herein.
Embodiment 1
Pichia spp YPD-YL2 fermented liquid obtains the making of solid deodorizer: after getting pichia spp YPD-YL2 fermented liquid 100ml and 200g wood chip Homogeneous phase mixing, at 35 DEG C, carry out drying to gained wood chip, when being 60% to its water ratio, obtain solid deodorizer.Gained solid deodorizer is carried out deodorizing effect test according to method listed in above-mentioned experiment 3.
Embodiment 2
Be with the difference of embodiment 1, used carrier is diatomite, pichia spp YPD-YL2 fermented liquid 100ml and 100g wood chip Homogeneous phase mixing.
Embodiment 3
Be with the difference of embodiment 1: used carrier is wilkinite, pichia spp YPD-YL2 fermented liquid 100ml and 2000g wood chip Homogeneous phase mixing.The test result of embodiment 1 ~ 3 is listed in table 5.
The clearance result table of table 5 embodiment 1 ~ 3 pair of hydrogen sulfide
| Embodiment | 1 | 2 | 3 |
| Hydrogen sulfide stripping rate (%) | 56.2 | 56.4 | 57.3 |
From table 5, solid deodorizer made by pichia spp YPD-YL2 fermented liquid provided by the invention, and higher may be used for of the removal efficiency for hydrogen sulfide carries out deodorization process to hydrogen sulfide containing foul gas.
Embodiment 4
Composite fungus agent by pichia spp YPD-YL2 fermented liquid, lactobacillus and subtilis by biomass ratio be 2: 1: 1 mixing after obtain.Corn stalk powder 50g/L, wheat bran 20g/L is at least comprised, brown sugar 80g/L, tomato powder 40g/L in composite fungus agent liquid fermentation medium used.Gained composite fungus agent effect is detected, with reference to the step and method in experiment 4.
Embodiment 5
Be with the difference of embodiment 4: composite fungus agent by pichia spp YPD-YL2 fermented liquid, lactobacillus and subtilis by biomass ratio be 7: 3: 1 mix after obtain.Corn stalk powder 150g/L, wheat bran 70g/L is at least comprised, brown sugar 120g/L, tomato powder 60g/L in composite fungus agent liquid fermentation medium used.
Embodiment 6
Be with the difference of embodiment 4: in gained composite fungus agent, add the composite fungus agent coordinating preparation in by CN201310293372.3 disclosed in embodiment 1.
The deodorizing effect detected result of embodiment 4 ~ 6 gained composite fungus agent is listed in table 6.
Table 6 embodiment 4 ~ 6 gained composite fungus agent is to domestic refuse deodorizing sensory evaluation scores result table
| Handling duration (hour) | 0 | 4 | 8 | 24 |
| Embodiment 4 | 1 | 4 | 5 | 5 |
| Embodiment 5 | 1 | 4 | 5 | 5 |
| Embodiment 6 | 1 | 4 | 5 | 5 |
Note: +++ represent strong foul smell; ++ represent foul smell; + represent slight stench taste
From table 6, no matter composite fungus agent provided by the invention is used alone or mixes with other existing deodorizing microorganisms, all can produce good deodorizing effect to domestic refuse.
Therefore pichia spp YPD-YL2 provided by the invention may be used for the foul gas processed containing ammonia and hydrogen sulfide, reduces its odour concentration, improves the acceptable degree of human body and animal body.The beneficiating ingredient in rubbish also can be utilized simultaneously to maintain growth, shorten the interpolation frequency of reodorant.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a pichia spp, tool is characterised in that, described pichia spp is pichia spp YPD-YL2, and the Latin of described pichia spp is called Pichiasp., and preserving number is CCTCCM2015225.
2. the application of pichia spp described in claim 1 in biological deodorizing.
3. application according to claim 2, is characterized in that, described biological deodorizing comprises the deodorizing process carried out feed, rubbish or organic fertilizer.
4. a solid deodorizer, is characterized in that, the bacterium liquid containing, for example Pichi strain according to claim 1 is adsorbed on carrier and obtains.
5. solid deodorizer according to claim 4, is characterized in that, is 1: 1 ~ 20 to mix by the volume of described bacterium liquid and the mass ratio of described carrier.
6. a composite fungus agent, tool is characterised in that, comprises the fermented liquid of pichia spp YPD-YL2 as claimed in claim 1, lactobacillus and subtilis.
7. composite fungus agent according to claim 6, is characterized in that, described pichia spp YPD-YL2 fermented liquid, lactobacillus and subtilis are 2 ~ 7: 1 ~ 3: 1 mixing by biomass ratio.
8. composite fungus agent according to claim 7, is characterized in that, described pichia spp YPD-YL2 fermented liquid, lactobacillus and subtilis are 3: 2: 1 mixing by biomass ratio.
9. the composite fungus agent according to any one of claim 6 ~ 8, it is characterized in that, corn stalk powder 50 ~ 150g/L, wheat bran 20 ~ 70g/L is at least comprised, brown sugar 80 ~ 120g/L, tomato powder 40 ~ 60g/L in described composite fungus agent liquid fermentation medium used.
10. the application of the composite fungus agent according to any one of claim 6 ~ 9 in deodorizing.
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