CN108486017A - A kind of garden waste degradation bacterial agent - Google Patents
A kind of garden waste degradation bacterial agent Download PDFInfo
- Publication number
- CN108486017A CN108486017A CN201810408825.5A CN201810408825A CN108486017A CN 108486017 A CN108486017 A CN 108486017A CN 201810408825 A CN201810408825 A CN 201810408825A CN 108486017 A CN108486017 A CN 108486017A
- Authority
- CN
- China
- Prior art keywords
- parts
- garden waste
- degradation
- bacterium
- bacterial agent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
- 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
- C12N1/14—Fungi; Culture media therefor
Abstract
The present invention provides a kind of garden waste degradation bacterial agents, including microbial inoculum and basic culture solution, bacterium group includes Promicromonospora, Burkholderia, dredges the thermophilic hyphomycete of cotton like, Hayes bacterium and streptomycete, basic culture solution includes wheat bran, starch, xylose, beef extract, ammonium nitrate and sterile water, and the weight ratio of bacterium group and basic culture solution is 1：1.5‑1：2.This programme has the beneficial effect that：Garden waste is carried out after fermentation using the degradation bacterial agent of this programme, the degradation rate of hemicellulose, cellulose and lignin is all remarkably higher than a certain bacterium of exclusive use；The present invention program adds sugared degradation bacteria Hayes bacterium and streptomycete, can other polysaccharide materials in garden waste be carried out efficient degradation, promote whole fermentation process.The composite degradation microbial inoculum of the present invention, lignin degradation are efficient；It not only economizes the land resource, improves soil and ecology, alleviates environmental pollution, the matrix after degradation can also be turned waste into wealth, realize resource reutilization.
Description
Technical field
The invention belongs to garden waste process fields, and in particular to a kind of garden waste degradation bacterial agent.
Background technology
As city's green areas constantly expands, the fast development of Urban Greening and people require ecological environment
It improves, the yield of garden waste increases sharply and rises year by year.But most wastes are all not carried out dataization profit
With the overwhelming majority, equally using processing modes such as burning, landfills, is not only brought serious environmental problem, caused with house refuse
The wasting of resources can also increase garbage disposal cost, not meet the requirement of ecological environment.Most of garden wastes contain abundant
Organic matter, some areas make garden waste form matrix, answer again with reference to the composting mode of agricultural wastes, by compost
For gardens and urban afforestation, the pollution to environment was both reduced, had also been economized on resources, and had been a kind of disposition side of suitable sustainable development
Formula.
Garden waste main body is plant tissue, and main constituents have cellulose, lignin and hemicellulose, seldom contain
There is mankind's house refuse ingredient, more rich lignin is contained compared to agricultural wastes.These lignin are primarily present in plant
In the micro-pipe tissue of object cell wall, lignin molecule chain aggregation at microfibril be interweaved and inlay arrangement form crystalline phase and non-
Crystalline phase.This exquisite closely crystal structure has superpower defense reaction, chemical reagent or biological enzyme to be difficult to effective contact table
Face acts on therewith, so be difficult to be degraded.Composting process is mainly driven by microbiologic population therein, and multiple-microorganism is needed
Common cooperation is completed, and the leading microorganism of different compost waste degradations is also different.Due to the plant tissue of garden waste
With higher structural heterogeneity, higher silicone content and content of lignin, in the prior art about agricultural or house refuse
Compost treatment mode be simply not proposed to garden waste.Therefore the microbial bacteria for capableing of efficient degradation cellulose is filtered out
Kind, and the composite degradation microbial inoculum suitable for garden waste is researched and developed, the processing and environmental protection for garden waste have weight
Want meaning.
Invention content：
Aiming at the deficiencies in the prior art, the present invention provides a kind of garden waste degradation bacterial agent, can efficiently drop
Solve lignin so that garden waste is re-applied by compost and biodegradation in gardens and urban afforestation.
In order to achieve the above object, the present invention adopts the following technical scheme that：A kind of garden waste degradation bacterial agent, including bacterium group
And basic culture solution, bacterium group include Promicromonospora (Bacillus mojavensis), Burkholderia (Burkholderia
Sp.), dredge the thermophilic hyphomycete of cotton like (Thermomyces lanuginosus), Hayes bacterium (Shigella Castellani) and
Streptomycete (streptomyces), basic culture solution include wheat bran, starch, xylose, beef extract, ammonium nitrate and sterile water,
Bacterium group and the weight ratio of basic culture solution are 1：1.5-1：2.
Preferably, bacterium group includes following component in parts by weight in above-mentioned garden waste degradation bacterial agent：Promicromonospora
40-55 parts, 10-25 parts of Burkholderia, dredge cotton like is hyphomycete 15-25 parts thermophilic, 5-10 parts of Hayes bacterium, 5-10 parts of streptomycete.
Preferably, in above-mentioned garden waste degradation bacterial agent, basic culture solution pH value is 7.0-7.2, beef extract and sulfuric acid
Ammonium forms the mixture that mass fraction is 0.5-1%.
Preferably, in above-mentioned garden waste degradation bacterial agent, basic culture solution includes following component in parts by weight：Wheat
25-40 parts of 30-35 parts of wheat bran, 5-15 parts of starch, 5-15 parts of xylose, beef extract and ammonium nitrate mixture, 25-55 parts of sterile water.
Preferably, in above-mentioned garden waste degradation bacterial agent, garden waste degradation bacteria group includes as follows in parts by weight
Component：50-55 parts of Promicromonospora, 15-25 parts of Burkholderia, dredge cotton like is hyphomycete 20-25 parts thermophilic, 5-10 parts of Hayes bacterium,
5-10 parts of streptomycete.
Preferably, in above-mentioned garden waste degradation bacterial agent, basic culture solution includes following component in parts by weight：Wheat
35 parts of 35 parts of wheat bran, 10 parts of starch, 10 parts of xylose, beef extract and ammonium nitrate mixture, 35-50 parts of sterile water.
Preferably, in above-mentioned garden waste degradation bacterial agent, garden waste degradation bacteria group includes as follows in parts by weight
Component：50 parts of Promicromonospora, dredges 25 parts of the thermophilic hyphomycete of cotton like, 10 parts of Hayes bacterium, 10 parts of streptomycete at 20 parts of Burkholderia.
The present invention also provides a kind of processing methods using garden waste degradation bacterial agent degradation garden waste, including with
Lower step：S1, garden waste carry out just processing, are substantially shredded and are shakeout, 5-15 centimetres of thickness；S2, gardens are discarded
The microbial inoculum and basic culture solution of object degradation bacterial agent are uniformly mixed；S3, it by mixed degradation bacterial agent but broadcasts sowing and is seeded to gardens and gives up
The weight ratio of gurry surface, degradation bacterial agent and garden waste is 1：200-1：300；S4, it the gardens after bacterium will be connect gives up
Gurry stirs evenly, and is covered with plastic foil and ferments, and keeps 20-35 DEG C of temperature, and processing after a week is completed.
The principles and methods of the present invention program are as follows：
Cellulose be by glucose group at macromolecular polysaccharide, be the main component of plant cell wall.Hemicellulose be by
Heteromultimer that several different types of monosaccharide are constituted, including xylose, Ah 's sugar, mannose and galactolipin etc..Lignin is
It is only second to the organic matter the abundantest of cellulose in nature, does not contain the repetitive unit of facile hydrolysis.In general timber, fiber
Element accounts for 40~50%, also 10~30% hemicellulose and 20~30% lignin.Microorganism by generate cellulase,
The biology enzyme such as lignin peroxidase, is hydrolyzed into small-molecule substance by cellulose and lignin, completes degradation process.Single
Component of enzyme system is unable to final degradation of the complete independently to natural lignin's substrate, is glucose etc. natural lignin's degradation of substrates
Monosaccharide, it is necessary to could be completed under the collective effect of a few fiber element component of enzyme system.It can improve body using composite bacteria agent one
The vigor and stability of various enzyme components in system；Two come by other related with lignin degradation in abundant co-culture system
The small molecule active factor and the type of organized enzyme are realized.
Degradation bacterial agent in the present invention program includes microbial inoculum and basic culture solution, and bacterium group includes Promicromonospora, primary kirschner
Bacterium dredges the thermophilic hyphomycete of cotton like, Hayes bacterium and streptomycete.Find that Promicromonospora coordinates primary gram by the complicated experiment of inventor
Salmonella, the two are very high to the degradation capability and efficiency of lignin.And hemicellulose present in natural lignin imitates degradation
Rate has important influence, experiment to find that during lignin degradation, the wooden poly oligosaccharide of high concentration can inhibit cellulase
Activity, and it is very efficient hemicellulose degradation bacterium to dredge the thermophilic hyphomycete of cotton like, is added and dredges the thermophilic hyphomycete of cotton like on the one hand drop
The hemicellulose in garden waste is solved, while the degradation efficiency of cellulose-degrading bacteria can also be improved.Namely in natural lignin
Degradation process in, synergistic effect has occurred between hemicellulase and cellulase, promotes cellulose and hemicellulose
Conversion ratio.Hayes bacterium and streptomycete are sugared degradation bacterias, and have abundant sugared content in the plant tissue of garden waste, are added and congratulate
Salmonella and streptomycete can accelerate entire compost degradation treatment process.
Because in the compost composting process of garden waste, need to form stable microbiologic population's knot as early as possible
Structure, to complete degradation treatment as early as possible.The present invention microbial inoculum further include wheat bran, starch, xylose, beef extract, ammonium nitrate and
The weight ratio of the basic culture solution of sterile water composition, bacterium group and basic culture solution is 1：1.5-1：2.Shallow lake in basic culture solution
Powder, beef extract, ammonium nitrate and sterile water help the various bacterium in bacterium group preserve, maintenance is active, are to aid in sprinkle to be discarded in gardens
The auxiliary agent quickly bred behind object surface.Xylose and wheat bran in basic culture solution are the excellent derivants of zytase,
Can acute activation and induction the present invention bacterium group carry out degradation relevant enzyme secretion.
The present invention program's has the beneficial effect that：It is demonstrated experimentally that using this programme degradation bacterial agent to garden waste into
After fermentation, the degradation rate of hemicellulose, cellulose and lignin is all remarkably higher than a certain bacterium of exclusive use to row；The present invention
Scheme adds sugared degradation bacteria Hayes bacterium and streptomycete, can efficiently be dropped other polysaccharide materials in garden waste
Solution promotes whole fermentation process.The present invention finds Suitable strains combination by experiment and prepares composite degradation microbial inoculum, significantly improves
The degradation efficiency of lignin；It not only economizes the land resource, improves soil and ecology, alleviates environmental pollution, it can also will be after degradation
Matrix turn waste into wealth, realize resource reutilization.
Description of the drawings
Fig. 1 is that five kinds of bacterial strains of experimental example of the present invention produce the analysis chart of filter paper enzyme activity；
Fig. 2 is that five kinds of bacterial strains of experimental example of the present invention produce the analysis chart of CMC enzyme activity；
Fig. 3 is that five kinds of bacterial strains of experimental example of the present invention produce the analysis chart of circumscribed enzyme activity.
Specific implementation mode
The selected embodiment of the present invention is illustrated referring now to Figure of description, those skilled in the art are according to the sheet of the disclosure
Subordinate's explanation of the embodiment of invention is merely exemplary, the scheme being not meant to limit the present invention.
Embodiment 1
A kind of garden waste degradation bacterial agent, including bacterium group and basic culture solution, bacterium group include 40 parts of Promicromonospora, primary
10 parts of kirschner bacterium dredges 15 parts of the thermophilic hyphomycete of cotton like, 5 parts of Hayes bacterium, 5 parts of streptomycete；Basic culture solution includes in parts by weight
Following component：25 parts of 30 parts of wheat bran, 5 parts of starch, 5 parts of xylose, beef extract and ammonium nitrate mixture, 25 parts of sterile water, ox
Meat extract and ammonium sulfate form the mixture that mass fraction is 0.5%；Bacterium group and the weight ratio of basic culture solution are 1：1.5；Basis
Medium pH value is 7.0.
Embodiment 2
A kind of garden waste degradation bacterial agent, including bacterium group and basic culture solution, bacterium group include such as the following group in parts by weight
Point：55 parts of Promicromonospora, dredges 20 parts of the thermophilic hyphomycete of cotton like, 8 parts of Hayes bacterium, 8 parts of streptomycete at 15 parts of Burkholderia；Basis
Culture solution includes following component in parts by weight：32 parts of wheat bran, 15 parts of starch, 15 parts of xylose, beef extract and ammonium nitrate are mixed
30 parts of object, 40 parts of sterile water are closed, beef extract and ammonium sulfate form the mixture that mass fraction is 1%；Bacterium group and basic culture solution
Weight ratio be 1：1.8；Basic culture solution pH value is 7.0.
Embodiment 3
A kind of garden waste degradation bacterial agent, including bacterium group and basic culture solution, bacterium group include such as the following group in parts by weight
Point：50 parts of Promicromonospora, dredges 25 parts of the thermophilic hyphomycete of cotton like, 10 parts of Hayes bacterium, 10 parts of streptomycete at 20 parts of Burkholderia；Base
Plinth culture solution includes following component in parts by weight：35 parts of wheat bran, 10 parts of starch, 10 parts of xylose, beef extract and ammonium nitrate
35 parts of mixture, 50 parts of sterile water, beef extract and ammonium sulfate form the mixture that mass fraction is 1%；Bacterium group is cultivated with basis
The weight ratio of liquid is 1：2；Basic culture solution pH value is 7.2.
The present invention also provides a kind of methods of garden waste degradation, specifically include following steps：S1, garden waste into
Row just processing, is substantially shredded and is shakeout, 5-15 centimetres of thickness；S2, by the microbial inoculum of garden waste degradation bacterial agent and basis
Culture solution is uniformly mixed；S3, it by mixed degradation bacterial agent but broadcasts sowing and is seeded to garden waste surface, degradation bacterial agent and gardens
The weight ratio of waste is 1：200-1：300；S4, it the garden waste after bacterium will be connect stirs evenly, be covered with plastic foil
It ferments, keeps 20-35 DEG C of temperature, processing after a week is completed.
The detection method of three kinds of cellulose degradation experimental datas：The degradation bacterial agent in above-described embodiment 1,2 and 3 is taken, and
The hereinafter degradation bacterial agent in comparative example 1,2 and 3, carries out degradation experiment as steps described below respectively：S1, culture medium is made：It receives
The processing sample as garden waste such as hedgerow branches and leaves, camphor tree fallen leaves after collection trimming, a processing sample is 20g, is crushed
It is mixed afterwards with the basic culture solution in degradation bacterial agent, solid-state fermentation culture medium is made after 12l DEG C of sterilizing 20min；S2, preparation bacterium are outstanding
Supernatant liquid：Bacteria suspension in Example 1,2 and 3 and comparative example 1,2 and 3 makes its concentration control 10 by adjusting turbidity7A/
1ml bacterium solutions are added in the ml orders of magnitude, each handle in sample culture medium；S3, inoculation：By bacterial suspension inoculation to each culture medium, often
Kind bacteria suspension is inoculated with 3 culture mediums respectively, and the culture medium of 3 blank is arranged as a contrast；S4,25-30 DEG C of temperature, place are kept
Reason takes out the content for measuring hemicellulose, cellulose and lignin in each processing culture medium, the survey of every 3 culture mediums after a week
Fixed number value is averaged, and calculates degradation rate.
Comparative example 1
A kind of garden waste degradation bacterial agent, including bacterium group and basic culture solution, bacterium group include such as the following group in parts by weight
Point：20 parts of Burkholderia dredges 25 parts of the thermophilic hyphomycete of cotton like, 10 parts of Hayes bacterium, 10 parts of streptomycete；Basic culture solution is by weight
Number includes following component：35 parts of wheat bran, 10 parts of starch, 10 parts of xylose, beef extract and 35 parts of ammonium nitrate mixture, sterile water
50 parts, beef extract and ammonium sulfate form the mixture that mass fraction is 1%；Bacterium group and the weight ratio of basic culture solution are 1：2；Base
Plinth medium pH value is 7.2.
Comparative example 2
A kind of garden waste degradation bacterial agent, including bacterium group and basic culture solution, bacterium group include such as the following group in parts by weight
Point：50 parts of Promicromonospora dredges 25 parts of the thermophilic hyphomycete of cotton like, 10 parts of Hayes bacterium, 10 parts of streptomycete；Basic culture solution is by weight
Number includes following component：It is 35 parts of 35 parts of wheat bran, 10 parts of starch, 10 parts of xylose, beef extract and ammonium nitrate mixture, sterile
50 parts of water, beef extract and ammonium sulfate form the mixture that mass fraction is 1%；Bacterium group and the weight ratio of basic culture solution are 1：2；
Basic culture solution pH value is 7.2.
Comparative example 3
A kind of garden waste degradation bacterial agent, including bacterium group and basic culture solution, bacterium group include such as the following group in parts by weight
Point：50 parts of Promicromonospora, 20 parts of Burkholderia, 10 parts of Hayes bacterium, 10 parts of streptomycete；Basic culture solution includes in parts by weight
Following component：35 parts of 35 parts of wheat bran, 10 parts of starch, 10 parts of xylose, beef extract and ammonium nitrate mixture, 50 parts of sterile water,
Beef extract and ammonium sulfate form the mixture that mass fraction is 1%；Bacterium group and the weight ratio of basic culture solution are 1：2；Basis training
Nutrient solution pH value is 7.2.
The degradation rate of the three kinds of celluloses of different embodiment and comparative examples pair of table 1
| Microbial inoculum classification | Hemicellulose degradation rate | Cellulose degradation rate | Lignin degradation rate |
| Embodiment 1 | 63.1% | 77.2% | 74.4% |
| Embodiment 2 | 66.3% | 80.5% | 72.1% |
| Embodiment 3 | 72.7% | 86.4% | 77.9% |
| Comparative example 1 | 68.4% | 53.5% | 61.1% |
| Comparative example 2 | 65% | 69.4% | 64.3% |
| Comparative example 3 | 37.9% | 82.3% | 59.9% |
By the statistical result of upper table 1 it is found that after fermentation, microbial inoculum ingredient in embodiment 1,2 and 3 to hemicellulose,
The degradation rate of cellulose and lignin is higher than comparative example 1,2 and 3.This illustrates the combination energy of various mushrooms in the present invention program
The degradation rate of larger higher hemicellulose, cellulose and lignin.Wherein Promicromonospora is substantially carried out cellulose and wooden
The degradation of element, is not added Promicromonospora in comparative example 1, thus after fermentation in its culture medium cellulose and lignin drop
Solution rate only has 53.5% and 61.1%；Burkholderia is substantially carried out the degradation of cellulose and lignin, and primary is not added in comparative example 2
Kirschner bacterium, therefore the degradation rate of cellulose and lignin only has 69.4% and 64.3% in its culture medium after fermentation；Dredge cotton
Shape is thermophilic, and hyphomycete is substantially carried out the degradation of hemicellulose, therefore the thermophilic hyphomycete of thin cotton like is not added in comparative example 3, therefore ferments
After in its culture medium the degradation rate of hemicellulose there was only 37.9%.
Experimental example
The cellulose degradation capability analysis of Promicromonospora：The single bacterium colony of picking Promicromonospora is inoculated in carboxylic with point connection
On sodium carboxymethylcellulose pyce solid medium, repeated inoculation 3.After cultivating 3d, the rigid of 10mL 1mg/mL is added into each tablet
Arnotto solution, Congo red dye liquor is abandoned after standing dyeing 15min, 1mol/L NaCl solutions washing flat board is then added 3~5 times,
Periphery of bacterial colonies is observed whether there is or not transparent circle appearance, illustrates that the bacterial strain can generate cellulase if occurring, and use right-angled intersection
Method measures colony diameter (d) and transparent diaphragm diameter (D), according to the cellulose of transparent circle bacterial strain compared with the diameter of bacterium colony is than coming
Degradation capability, i.e. cellulase relative activity=D/d.In the experimental example, the diaphragm diameter D values of Promicromonospora are 0.25cm,
Colony diameter d values are 0.12cm, and enzyme relative activity (D/d) value is 2.08, show that this kind of colony morphological observation is easy, and it is fine to degrade
The ability for tieing up element is stronger.Promicromonospora is Gram-positive, not antiacid, catalase positive.Aerobic, medium temperature, change can have
Machine nutrition.Branch mycelia gos deep into culture medium, fragments into different size of segment, and chlamydospore shape base silk is fractured into corynebacterium diphtheriae shape
With coccus shape cell, some generates single spore on Ji Si or short handle.
Cellulase (β-Isosorbide-5-Nitrae-glucan -4- glucan hydrolases) is one kind of enzyme, in decomposition of cellulose from biology
Catalytic action, can be by cellulose decomposition at glucose.Cellulase is the total of the glucogenic one group of enzyme of degraded cellulose
Claim, its not instead of monomeric enzyme plays the multicomponent enzyme system of synergistic effect, is a kind of complex enzyme, mainly by circumscribed 1,4 beta-glucanase,
The compositions such as Endo-β-glucanase and beta-glucosidase.
Principle based on the Congo red plate screening High Cellulase Production bacterial strains of CMC-Na in above-described embodiment.Congo red method
(congo red) is that a kind of comparison generally acknowledged at present efficiently separates method.There is β -1,4-D- pyranoid forms in Congo red and structure
The polysaccharide of glucose has strong interaction, with β -1, lactose-Portugal's mannose polymerization of 3-D- glucans and some hemicelluloses
There is apparent interaction in sugar.It is rigid that polysaccharide hydrolysis object after Congo red same cellulose hydrolysis forms the strong polysaccharide-of color
Arnotto complex precipitate has apparent indicative function to identification polysaccharide hydrolysis object, detached for the Congo red culture medium of cellulose-,
Screen fibre element decomposer provides theoretical foundation.If bacterium colony itself can generate cellulase, the cellulose family around it
After substance is hydrolyzed, the red Polysaccharides-congo red complex that the Congo red polysaccharide product with after hydrolysis is formed is molten through sodium chloride
Periphery of bacterial colonies lighter forms transparent circle after liquid washes, and the strong red material of color is concentrated mainly on bacterium colony and does not grow
Region.It is Congo red can with cellulose formed red complex, but and get along well hydrolysis after cellobiose or glucose occur it is this
Reaction.Therefore when being added Congo red in the culture medium containing cellulose, it is Congo red can be with the formation of the cellulose in culture medium
Red complex, if after the cellulose in culture medium is decomposed by cellulase, the compound of Congo red-cellulose just can not shape
At ultimately forming red precipitate, color is strong, thick and heavy, can be chlorinated sodium solution elution, will appear in culture medium with cellulose point
The high-visible transparent circle centered on bacterium is solved, according to whether generating transparent circle carrys out screen fibre element decomposer.Transparent circle it is big
Small and cellulase activity is at positive correlation.It is test method with the advantages of cellulose Screening of Media cellulase production bacterium
Simply, the screening period is short, can be carried out just to the cellulase-producing ability of bacterium by hydrolyzing the production of transparent circle
The judgement of step.The morning that hydrolysis transparent circle occurs illustrates that strain enzyme-producing speed is fast, the bigger fibre for illustrating bacterial strain and generating of hydrolysis transparent circle
The plain enzyme of dimension is more, active higher.In test according to the time of occurrence and diameter for hydrolyzing transparent circle in cellulose culture medium
Size, the enzymatic productivity for judging bacterial strain that can be rough reduce the difficulty of screening operation a little to a certain extent.
The diversity of cellulase Function Class depends on the structural complexity of cellulosic substrate, but all follows synergistic effect
Mechanism is exactly beta-glucosidase, exoglucanase, endoglucanase generates synergistic effect becomes Portugal by cellulose degradation
Grape sugar：Endoglucanase is turned off the amorphous domain inside fibrination sugar chain, and makes a breach, then random hydrolysis
β-Isosorbide-5-Nitrae-glycosidic bond generates the reproducibility of different length and the fibrination sugar-chain end of irreducibility.Exoglucanase,
Cellobiohydrolase is can be described as, effect is the free terminal in these cellulose chains, and releases glucose or fiber
Disaccharides.In addition, exoglucanase hydrolysis can make the structure in cellulose crystallite region become loose so that be conducive to endo-glucanase
The effect of enzyme, to the end beta-glucosidase hydrolysis fiber disaccharides and oligosaccharides generate glucose, to release them to circumscribed Portugal
The feedback inhibition of dextranase and endoglucanase so that both enzymes can significantly more efficient effect.
The interpretation of result of different enzyme activity：
1.1 glucose standard curve
Standard glucose solution：Glucose configures 1.037g/ml concentration Portugal after 108 DEG C of drying in oven 30min~1h
Grape sugar juice, then 0ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml, 1.4ml glucose solution is taken to steam respectively
Distilled water is diluted to 2ml, and 3mlDNS reagent boiling water bath 10min are added, and is terminated react with cold water immediately, separately used 0ml, be in wavelength
Absorbance value (OD values) is measured under 540nm, absorbance value draws out Portugal as ordinate using glucose content as abscissa
Grape Standard for Sugars curve and fitting equation.Enzyme activity is by the definition of international unit：1 μm of ol grape is generated per min catalyzing hydrolysis
The enzyme amount of sugar is an enzyme activity unit IU.
The measurement of 1.2 filter paper enzyme activities
It takes 1ml bacterium solutions to centrifuge 10min in the centrifuge of 6000r, discards precipitation, filter paper substrate solution is added.At 50 DEG C
1h is reacted in thermostat water bath.3mlDNS solution is added after reaction, is put in boiling water bath and boils 10min, terminated immediately with cold water
Reaction.Absorbance value (OD values) is measured in the case where wavelength is 540nm.The activity of Filter paperlyase is calculated according to glucose standard curve.It is right
According to for bacterium solution boiling water 10min, cold water cools down immediately.Each sample do three it is parallel.
The measurement of 1.3CMC enzyme activity (endoglucanase)
It takes 1ml bacterium solutions to centrifuge 10min in the centrifuge of 6000r, discards precipitation, CMC substrate solutions are added.In 50 DEG C of perseverances
30min is reacted in warm water bath.3mlDNS solution is added after reaction, is put in boiling water bath and boils 10min, terminated immediately with cold water
Reaction.Absorbance value (OD values) is measured in the case where wavelength is 540nm.The activity of Filter paperlyase is calculated according to glucose standard curve.It is right
According to for bacterium solution boiling water 10min, cold water cools down immediately.Each sample do three it is parallel.
The measurement of 1.4 microcrystalline cellulose enzyme activity (exoglucanase)
It takes 1ml bacterium solutions to centrifuge 10min in the centrifuge of 6000r, discards precipitation, microcrystalline cellulose substrate solution is added.
2h is reacted in 50 DEG C of thermostat water baths.3ml DNS solution is added after reaction, is put in boiling water bath and boils 10min, it is vertical with cold water
Terminate reaction.Absorbance value (OD values) is measured in the case where wavelength is 540nm.The work of Filter paperlyase is calculated according to glucose standard curve
Property.Control is bacterium solution boiling water 10min, and cold water cools down immediately.Each sample do three it is parallel.
The canonical plotting for formulating the glucose test of this experimental example show that the equation of the standard curve of glucose experiment is
It, will be acquired when y=7.9736x+0.0002 (x OD=540, y are standard sample concentration of glucose) sample glucose content
The absorption value of light be updated in equation, acquire glucose content in sample.
Fig. 1 is that five kinds of bacterial strains of this experimental example produce the analysis chart of filter paper enzyme activity, wherein this five kinds of bacterial strains are respectively：A is former
Micromonospora, B are Burkholderias, and C is the short stalk bacterium of budding, and D is production flavomycoin, and E is Promicromonospora (similarly hereinafter).Bacterial strain is transferred
Enter in producing enzyme inducing culture, measures filter paper enzyme activity.Filter paper is more loose as substrate structure, is that the degree of polymerization and crystallinity all occupy
Medium fibrous material is easy to be degraded by restriction endonuclease and excision enzyme simultaneously, then resolves into the reduction such as glucose by the sweet enzyme of glucose
Sugar, reflection be three fermentoid component of cellulase synergistic effect total enzyme activity.From figure 2 it can be seen that the enzyme after connecing bacterial strain
Vigor is as the variation of time is in the trend that gradually rises, and when 3d, enzyme activity reaches peak.A, E and D are in third day yield of enzyme
Reach 100U or more.After incubation time is more than 3d, enzyme activity begins to decline.This illustrates that Promicromonospora is filtered with stronger production
The ability of paper enzyme activity.
Fig. 2 is that five kinds of bacterial strains of this experimental example produce the analysis chart of CMC enzyme activity.Bacterial strain is transferred into producing enzyme inducing culture
In, measure CMC (hydroxymethyl cellulose) enzyme activity.Since excision enzyme is high to the specificity of cellulose chain, and the specificity of restriction endonuclease
Relatively low, when degrading CMC, what is mainly reflected is the vigor of restriction endonuclease.From figure 3, it can be seen that increasing of the enzyme activity with the time
Add and rise, when 3d, enzyme activity reaches peak.A, C and E produced CMC enzyme activity at the 3d days and reach 140U or more, wherein C bacterium
Strain reaches 150U or more in 3d yield of enzyme.After incubation time is more than 3d, enzyme activity begins to decline.This illustrates that Promicromonospora has
There is the ability of stronger production CMC enzyme activity.
Fig. 3 is that five kinds of bacterial strains of this experimental example produce the analysis chart of circumscribed enzyme activity.Bacterial strain is connected to the induction for capableing of producing enzyme
In culture medium, from figure 3, it can be seen that enzyme activity rises as time increases, in 2d, enzyme activity reaches most by B, C and E
High level, A and D reach maximum value in 3d.Later, enzyme activity begins to decline.This explanation is transferred in the bacterial strain of cellulose-degrading bacteria
When the initial stage of bacterium solution, the active rate of rise of the enzyme of the bacterial strain B and C of cellulose-degrading bacteria is relatively high, and three plants other
The vigor rate of rise of the enzyme of the bacterial strain of cellulose-degrading bacteria is relatively slow.This illustrates that Promicromonospora has stronger production circumscribed
The ability of enzyme activity.
Finally it should be noted that above example is merely to illustrate the technical solution of the application rather than to its protection domain
Limitation, although the application is described in detail with reference to above-described embodiment, the those of ordinary skill in the field should
Understand：Those skilled in the art read the specific implementation mode of application can still be carried out after the application various changes, modification or
Equivalent replacement, but the above change, modification or equivalent replacement, the application wait authorizing or the claim of issued for approval protects model
Within enclosing.
Claims (8)
1. a kind of garden waste degradation bacterial agent, including bacterium group and basic culture solution, which is characterized in that the bacterium group includes former small
Sporangium (Bacillus mojavensis), dredges the thermophilic hyphomycete of cotton like at Burkholderia (Burkholderia sp.)
(Thermomyces lanuginosus), Hayes bacterium (Shigella Castellani) and streptomycete (streptomyces),
The basic culture solution includes wheat bran, starch, xylose, beef extract, ammonium nitrate and sterile water, the bacterium group and the basis
The weight ratio of culture solution is 1：1.5-1：2.
2. a kind of garden waste degradation bacterial agent according to claim 1, which is characterized in that the garden waste degradation
Bacterium group includes following component in parts by weight：Described Promicromonospora 40-55 parts, Burkholderia 10-25 parts described, the thin cotton
It is shape is thermophilic 15-25 parts of hyphomycete, 5-10 parts of the Hayes bacterium, streptomycete 5-10 parts described.
3. a kind of garden waste degradation bacterial agent according to claim 2, which is characterized in that the basic culture solution pH value
For 7.0-7.2, the beef extract and ammonium sulfate form the mixture that mass fraction is 0.5-1%.
4. a kind of garden waste degradation bacterial agent according to claim 3, which is characterized in that the basic culture solution is by weight
It includes following component to measure number：Described wheat bran 30-35 parts, starch 5-15 parts described, the xylose 5-15 parts described, beef
It is 25-40 parts of cream and ammonium nitrate mixture, sterile water 25-55 parts described.
5. a kind of garden waste degradation bacterial agent according to claim 4, which is characterized in that the garden waste degradation
Bacterium group includes following component in parts by weight：Described Promicromonospora 50-55 parts, Burkholderia 15-25 parts described, the thin cotton
It is shape is thermophilic 20-25 parts of hyphomycete, 5-10 parts of the Hayes bacterium, streptomycete 5-10 parts described.
6. a kind of garden waste degradation bacterial agent according to claim 5, which is characterized in that the basic culture solution is by weight
It includes following component to measure number：35 parts of the wheat bran, 10 parts of the starch, 10 parts of the xylose, the beef extract and nitre
It is 35 parts of sour ammonium mixture, sterile water 35-50 parts described.
7. a kind of garden waste degradation bacterial agent according to claim 5, which is characterized in that the garden waste degradation
Bacterium group includes following component in parts by weight：50 parts of the Promicromonospora, 20 parts of the Burkholderia, the thin cotton like is thermophilic
25 parts of hyphomycete, 10 parts of the Hayes bacterium, 10 parts of the streptomycete.
8. a kind of processing using the garden waste degradation bacterial agent degradation garden waste as described in any one of claim 1 to 7
Method includes the following steps：S1, garden waste carry out just processing, are substantially shredded and are shakeout, 5-15 centimetres of thickness；S2、
The microbial inoculum and basic culture solution of garden waste degradation bacterial agent are uniformly mixed；S3, it by mixed degradation bacterial agent but spreads
It broadcasts and is seeded to garden waste surface, the weight ratio of the degradation bacterial agent and garden waste is 1：200-1：300；
S4, it the garden waste after bacterium will be connect stirs evenly, be covered with plastic foil and ferment, keep 20-35 DEG C of temperature, handle after a week
It completes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810408825.5A CN108486017A (en) | 2018-04-27 | 2018-04-27 | A kind of garden waste degradation bacterial agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810408825.5A CN108486017A (en) | 2018-04-27 | 2018-04-27 | A kind of garden waste degradation bacterial agent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108486017A true CN108486017A (en) | 2018-09-04 |
Family
ID=63314443
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810408825.5A Pending CN108486017A (en) | 2018-04-27 | 2018-04-27 | A kind of garden waste degradation bacterial agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108486017A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109777753A (en) * | 2019-01-24 | 2019-05-21 | 中南民族大学 | The micromonospora YG-1 bacterial strain and application thereof of one plant of production polysaccharide |
| CN110938550A (en) * | 2019-12-23 | 2020-03-31 | 山东大学 | Method for promoting thermobifida fusca to degrade agricultural wastes |
| CN111069237A (en) * | 2019-12-20 | 2020-04-28 | 武汉工程大学 | Medium-low grade phosphorite heap leaching method combining composite strain with waste biomass |
| CN112625921A (en) * | 2020-12-29 | 2021-04-09 | 中国科学院成都生物研究所 | Bacterium preparation for treating waste with high lignin content |
| CN115074295A (en) * | 2022-07-21 | 2022-09-20 | 华中农业大学 | Bacillus belgii growing by taking camphor as carbon source, high-temperature compost decomposing inoculant and application |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876754A (en) * | 2004-01-16 | 2013-01-16 | 诺维信股份有限公司 | Methods for degrading lignocellulosic materials |
| CA2859796A1 (en) * | 2011-12-19 | 2013-06-27 | Novozymes A/S | Processes and compositions for increasing the digestibility of cellulosic materials |
| WO2016026977A1 (en) * | 2014-08-22 | 2016-02-25 | Danmarks Tekniske Universitet | A process for producing a fermentation product from a lignocellulose-containing material |
| CN105567612A (en) * | 2016-03-05 | 2016-05-11 | 北京林业大学 | Preparation and application of garden waste compound degrading microbial agent |
| CN106947722A (en) * | 2017-04-28 | 2017-07-14 | 连云港中新污水处理有限公司 | Microbial bacterial agent and preparation method thereof and the application in sewage disposal |
| WO2018019707A1 (en) * | 2016-07-25 | 2018-02-01 | Metgen Oy | Method for lignin depolymerisation |
-
2018
- 2018-04-27 CN CN201810408825.5A patent/CN108486017A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876754A (en) * | 2004-01-16 | 2013-01-16 | 诺维信股份有限公司 | Methods for degrading lignocellulosic materials |
| CA2859796A1 (en) * | 2011-12-19 | 2013-06-27 | Novozymes A/S | Processes and compositions for increasing the digestibility of cellulosic materials |
| WO2016026977A1 (en) * | 2014-08-22 | 2016-02-25 | Danmarks Tekniske Universitet | A process for producing a fermentation product from a lignocellulose-containing material |
| CN105567612A (en) * | 2016-03-05 | 2016-05-11 | 北京林业大学 | Preparation and application of garden waste compound degrading microbial agent |
| WO2018019707A1 (en) * | 2016-07-25 | 2018-02-01 | Metgen Oy | Method for lignin depolymerisation |
| CN106947722A (en) * | 2017-04-28 | 2017-07-14 | 连云港中新污水处理有限公司 | Microbial bacterial agent and preparation method thereof and the application in sewage disposal |
Non-Patent Citations (2)
| Title |
|---|
| SANG MI CHOI等: "Characterization of Bacillus mojavensis KJS-3 for industrial applications", 《ARCH PHARM RES》 * |
| SAWSSEN HAJJI等: "Chitin extraction from crab shells by Bacillus bacteria. Biological activities of fermented crab supernatants", 《INT J BIOL MACROMOL》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109777753A (en) * | 2019-01-24 | 2019-05-21 | 中南民族大学 | The micromonospora YG-1 bacterial strain and application thereof of one plant of production polysaccharide |
| CN109777753B (en) * | 2019-01-24 | 2020-05-15 | 中南民族大学 | Micromonospora YG-1 strain for producing polysaccharide and application thereof |
| CN111069237A (en) * | 2019-12-20 | 2020-04-28 | 武汉工程大学 | Medium-low grade phosphorite heap leaching method combining composite strain with waste biomass |
| CN111069237B (en) * | 2019-12-20 | 2021-07-13 | 武汉工程大学 | Medium-low grade phosphorite heap leaching method combining composite strain with waste biomass |
| CN110938550A (en) * | 2019-12-23 | 2020-03-31 | 山东大学 | Method for promoting thermobifida fusca to degrade agricultural wastes |
| CN112625921A (en) * | 2020-12-29 | 2021-04-09 | 中国科学院成都生物研究所 | Bacterium preparation for treating waste with high lignin content |
| CN115074295A (en) * | 2022-07-21 | 2022-09-20 | 华中农业大学 | Bacillus belgii growing by taking camphor as carbon source, high-temperature compost decomposing inoculant and application |
| CN115074295B (en) * | 2022-07-21 | 2023-10-24 | 华中农业大学 | Bacillus bailii growing by taking camphor as carbon source, high-temperature composting and decomposing inoculant and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xia et al. | Cellulase production by solid state fermentation on lignocellulosic waste from the xylose industry | |
| CN108486017A (en) | A kind of garden waste degradation bacterial agent | |
| CN102559506B (en) | Penicillium oxalicum and application thereof | |
| CN101974436B (en) | Lignocellulose degrading bacteria and application thereof | |
| CN102453676B (en) | Straw starter and use of the straw starter in straw fermentation | |
| CN106631248A (en) | Production process for bioorganic fertilizer with high bacteria content | |
| CN104692866A (en) | Biological organic fertilizer | |
| CN104692868A (en) | Application of bioorganic fertilizer to soil improvement | |
| CN100393672C (en) | Active additive for promoting agricultural waste material compost decomposition and its application | |
| CN104692844A (en) | Preparation method of bioorganic fertilizer | |
| CN102174423B (en) | Bacillus licheniformis CH15 for degrading straws and bacterial agent thereof | |
| CN103160441B (en) | Preparation process for Penicillium oxalicum fermentation broth and application thereof in agriculture | |
| CN103045566A (en) | Method for producing cellulase by using induction and regulation of substrate | |
| CN109486881A (en) | A kind of fermentation medium and zymotechnique of kasugarnycin | |
| CN104109636B (en) | Aspergillus versicolor SD-3 and in the application prepared in chitin deacetylase | |
| CN109161495A (en) | A kind of composite bacteria agent of efficient degradation stalk cellulose | |
| CN103408335B (en) | Microbiological method based harmlessness treatment technology for agricultural wastes generated by grape cultivation | |
| CN103131639B (en) | Trichoderma longibrachiatum strain and application thereof | |
| CN102382771B (en) | Strain produced by beta-glucosidase and method for preparing Genipin therefrom | |
| CN110734322A (en) | Method for assisting fermentation of crop straws by alginate | |
| CN102268379B (en) | Trametes trogii and method for producing cellulase by using same | |
| CN113481111B (en) | Efficient biological straw fermentation inoculant and preparation method thereof | |
| CN108796027A (en) | A method of producing carotenoid | |
| CN106554224A (en) | A kind of high-efficiency fermenting type Hericium erinaceus (Bull. Ex Fr.) Pers. compost and preparation method thereof | |
| CN106047775A (en) | Stain of bacillus megatherium for producing cellulase and application method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180904 |
|
| RJ01 | Rejection of invention patent application after publication |