CN107793060A - A kind of method that false bacillus firmus DSM8715 improves regenerative bone material by concrete - Google Patents
A kind of method that false bacillus firmus DSM8715 improves regenerative bone material by concrete Download PDFInfo
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- CN107793060A CN107793060A CN201711056103.XA CN201711056103A CN107793060A CN 107793060 A CN107793060 A CN 107793060A CN 201711056103 A CN201711056103 A CN 201711056103A CN 107793060 A CN107793060 A CN 107793060A
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- bacillus firmus
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- C—CHEMISTRY; METALLURGY
- C04—CEMENTS; CONCRETE; ARTIFICIAL STONE; CERAMICS; REFRACTORIES
- C04B—LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE; CERAMICS; REFRACTORIES; TREATMENT OF NATURAL STONE
- C04B20/00—Use of materials as fillers for mortars, concrete or artificial stone according to more than one of groups C04B14/00 - C04B18/00 and characterised by shape or grain distribution; Treatment of materials according to more than one of the groups C04B14/00 - C04B18/00 specially adapted to enhance their filling properties in mortars, concrete or artificial stone; Expanding or defibrillating materials
- C04B20/02—Treatment
- C04B20/023—Chemical treatment
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- 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
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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
- Y02W30/50—Reuse, recycling or recovery technologies
- Y02W30/91—Use of waste materials as fillers for mortars or concrete
Abstract
The invention provides a kind of method that false bacillus firmus DSM8715 improves regenerative bone material by concrete：1) false bacillus firmus DSM8715 is mixed with Mineralized Culture liquid, obtains mixed-culture medium；The Mineralized Culture liquid includes the 10~45g/L propane sulfonic acid of 3 Cyclohexylamino 1,4~12g/L sodium lactate, 1~3g/L sodium nitrate, 20~40mmol/L calcium chloride, 0.5~1.5mmol/L magnesium sulfate, 0.08~0.18mmol/L potassium dihydrogen phosphate；2) regeneration aggregate is soaked in the mixed-culture medium that the step 1) obtains, strengthened regeneration aggregate.Regeneration aggregate is prepared using method provided by the invention, water absorption rate and crush index value substantially reduce, and the regeneration mortar compression strength prepared by the intensifying regenerating aggregate significantly increases.
Description
Technical field
The invention belongs to technical field of concrete, more particularly to a kind of false bacillus firmus DSM8715 to improve coagulation
The method of native regeneration aggregate.
Background technology
With urbanization progress faster in world wide, construction industry enters the high speed development stage.A large amount of old buildings are split
Remove, generate substantial amounts of building waste, wherein discarded concrete portion is maximum.Data shows, the whole world from 1991~
Between 10 years in 2000, discarded concrete (including from the underproof product of armored concrete factory) total amount is more than 1,000,000,000 tons.I
40,000,000 tons of calculating of building waste are removed by annual by state, wherein 34% is concrete block, then resulting discarded concrete is just
There are 13,600,000 tons, and with the quickening of economic construction paces, increased trend is presented.The discarded concrete of such flood tide is except processing
Expense is surprising outer, it is also necessary to takes substantial amounts of vacant lot storage, pollutes environment, waste arable land, turn into a big public hazards in city.
Discarded concrete is not only high-quality concrete aggregate, makees to gather materials with many advantages with waste concrete block, such as
After building disintegrates, concrete block and flour sand after high-quality crushing and screening can be as the regeneration of concrete, therefore utilize discarded
Building rubbish produces regeneration aggregate and prepares the important development direction that regeneration concrete is construction industry from now on.Discarded concrete
The processing method of most worthy is exactly that it is re-used production regeneration aggregate as renewable resource, is turned waste into wealth.
Regeneration aggregate is applied with before good economic benefit, social benefit, environmental benefit and the application of good market
Scape.In the prior art, or ball mill activating and regenerating aggregate is used so that the quality of regeneration aggregate greatly improves, available for producing
Reinforced concrete member；Or using the glacial acetic acid of 5% concentration and the hydrochloric acid solution of 3% concentration to regeneration aggregate processing；Or use
Cement and Separate Fine-grained Minerals slurry liquid (such as flyash, silica flour, siliceous waterproofing agent or calcium sulphoaluminate class swelling agent) are to regeneration aggregate
The processing such as immersion, dry.But because regeneration aggregate produces hole, crack etc. during broken or processing, regeneration aggregate is caused to inhale
Water rate is larger, causes regeneration concrete hydraulic performance decline, constrains the development in regeneration aggregate production regeneration concrete field.
The content of the invention
In view of this, in view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to which water absorption rate and pressure can be reduced by providing one kind
Broken index, improving the false bacillus firmus DSM8715 of mixed mortar compression strength improves the method for regenerative bone material by concrete.
The invention provides a kind of method that false bacillus firmus DSM8715 improves regenerative bone material by concrete, including such as
Lower step：
1) false bacillus firmus DSM8715 is mixed with Mineralized Culture liquid, obtains mixed-culture medium；The Mineralized Culture
The preparation raw material of liquid includes 10~45g/L 3- Cyclohexylamino -1- propane sulfonic acid, 4~12g/L sodium lactate, 1~3g/L nitric acid
Sodium, 20~40mmol/L calcium chloride, 0.5~1.5mmol/L magnesium sulfate, 0.08~0.18mmol/L potassium dihydrogen phosphate；
2) regeneration aggregate is soaked in the mixed-culture medium that the step 1) obtains, strengthened regeneration aggregate.
Preferably, the pH value of the Mineralized Culture liquid is 10.1~10.9.
Preferably, in the mixed-culture medium that the step 1) obtains, false bacillus firmus DSM8715 bacterium number for 1 ×
107~109cfu/L。
Preferably, step 1) the false bacillus firmus DSM8715 is obtained through microbiological culture media culture；
The microbiological culture media includes beef extract culture medium and 3- Cyclohexylamino -1- propane sulfonic acid buffer solutions；The beef
Cream culture medium is using water as solvent, including 2.5~4.5g/L beef extract and 8~15g/L peptone；The 3- Cyclohexylaminos-
1- propane sulfonic acid buffer solution includes 120~170g/L 3- Cyclohexylamino -1- propane sulfonic acid using water as solvent；The 3- Cyclohexylaminos-
The pH value of 1- propane sulfonic acid buffer solutions is 9~10.5；The body of the beef extract culture medium and 3- Cyclohexylamino -1- propane sulfonic acid buffer solutions
Product ratio is (70~95):(10~20).
Preferably, the culture is carried out under oscillating condition, and the frequency of the vibration is 120~180rpm；The culture
Temperature be 25~35 DEG C, the time of the culture is 8~48h.
Preferably, after microbiological culture media culture, centrifugation obtains thalline and sunk the false bacillus firmus DSM8715
Form sediment；The bacterial sediment is washed with the Mineralized Culture liquid 2~3 times；The rotating speed of the centrifugation is 5000~7000rpm, described
The time of centrifugation is 8~12 minutes.
As one of preferable technical scheme, the quality of the regeneration aggregate in the step 2) and the volume of mixed-culture medium
Than for (0.5~1.5) g:(15~25) ml.Now, the time of the step 2) immersion is preferably 15~30 days.
As the two of preferable technical scheme, in step 2), the regeneration aggregate also wraps after being soaked in mixed-culture medium
Include：The regeneration aggregate after the immersion is sprayed with Mineralized Culture liquid step 1) described, strengthened regeneration aggregate；Now, it is described
The time of immersion is 1~5h；The mode of the spray is intermittent shower, and 0.5~1.5h is sprayed every 10~12h, is regenerated per 1kg
The amount of aggregate spray mixed-culture medium is 1~3mL/min；The total time of the spray is 15~30d.
Beneficial effect：
The invention provides a kind of method that false bacillus firmus DSM8715 improves regenerative bone material by concrete, including such as
Lower step：1) false bacillus firmus DSM8715 is mixed with Mineralized Culture liquid, obtains mixed-culture medium；The Mineralized Culture
The preparation raw material of liquid includes 10~45g/L 3- Cyclohexylamino -1- propane sulfonic acid, 4~12g/L sodium lactate, 1~3g/L nitric acid
Sodium, 20~40mmol/L calcium chloride, 0.5~1.5mmol/L magnesium sulfate, 0.08~0.18mmol/L potassium dihydrogen phosphate；
2) regeneration aggregate is soaked in the mixed-culture medium that the step 1) obtains, strengthened regeneration aggregate.Regeneration aggregate is mixed
Close after being soaked in nutrient solution, microbes are attached in regeneration aggregate surface pore, using the composition in Mineralized Culture liquid, are passed through
It is metabolized calcium ion Ca therein2+It is converted into calcium carbonate CaCO3It is deposited in regeneration aggregate surface pore, so as to block Regenerated Bone
Expect surface pore, reduce the water absorption rate of regeneration aggregate.The regeneration aggregate being prepared using the method for the invention and microorganism
Before processing is compared, and water absorption rate reduces 15.1~28.3%.The regeneration mortar compression strength prepared by the intensifying regenerating aggregate
Increase by 6.9~23.4%.
Embodiment
The invention provides a kind of method of microbial augmentation regeneration aggregate, comprise the following steps：
1) false bacillus firmus DSM8715 is mixed with Mineralized Culture liquid, obtains mixed-culture medium；The Mineralized Culture
The preparation raw material of liquid includes 10~45g/L 3- Cyclohexylamino -1- propane sulfonic acid, 4~12g/L sodium lactate, 1~3g/L nitric acid
Sodium, 20~40mmol/L calcium chloride, 0.5~1.5mmol/L magnesium sulfate, 0.08~0.18mmol/L potassium dihydrogen phosphate；
2) regeneration aggregate is soaked in the mixed-culture medium that the step 1) obtains, strengthened regeneration aggregate.
The present invention is to the source of the false bacillus firmus DSM8715 (Bacillus pseudofirmus) without spy
It is different to limit.In the present invention, the false bacillus firmus DSM8715 is real by TU Delft Polytechnics Microlab
Room Henk Jonkers are tested to gift acquisition.
In the present invention, the false bacillus firmus DSM8715 preferably obtains through microbiological culture media culture；It is described micro-
Biological medium preferably includes beef extract culture medium and 3- Cyclohexylamino -1- propane sulfonic acid (CAPS) buffer solution.Beef extract culture medium
Nutritional ingredient is provided for the activation of strain, and CAPS buffer solutions are keeping the pH of microbiological culture media stable.
In the present invention, using water as solvent, the beef extract culture medium preferably includes 2.5~4.5g/L beef extract and 8
The beef extract of~15g/L peptone, more preferably 3.0~4.0g/L and 10~13g/L peptone, most preferably 3.53g/
L beef extract and 11.76g/L peptone.
In the present invention, used after the preferred sterilizing of the beef extract culture medium；The sterilizing is preferably high-temperature sterilization, described
The temperature of sterilizing is preferably 121 DEG C, and the time of the sterilizing is preferably 15min.The present invention is to the equipment of the sterilizing without spy
It is different to require, using those skilled in the art's conventional sterilant equipment, such as high-pressure steam sterilizing pan.
In the present invention, using water as solvent, the 3- Cyclohexylaminos -1- propane sulfonic acid buffer solutions preferably include 120~170g/
L 3- Cyclohexylamino -1- propane sulfonic acid, most preferably more preferably 140~160g/L, 147.5g/L.
In the present invention, the pH value of the 3- Cyclohexylaminos -1- propane sulfonic acid buffer solutions is preferably 9~10.5, more preferably
9.5~10.The method that the present invention adjusts pH value to 3- Cyclohexylamino -1- propane sulfonic acid buffer solution is not particularly limited, using ability
The Normal practice of field technique personnel.In the specific embodiment of the invention, preferably 3- Cyclohexylaminos -1- third is adjusted with NaOH
The pH value of sulfonate buffer.The NaOH is preferably the NaOH aqueous solution, and the concentration of the NaOH aqueous solution is preferably 1~10mol/
L, more preferably 5~7mol/l.
In the present invention, used after the preferred sterilizing of the 3- Cyclohexylaminos -1- propane sulfonic acid buffer solution；The sterilizing is preferably
High-temperature sterilization, the temperature of the sterilizing is preferably 121 DEG C, and the time of the sterilizing is preferably 15min.The present invention is to the sterilizing
Equipment there is no particular/special requirement, using those skilled in the art's conventional sterilant equipment, such as high-pressure steam sterilizing pan.
In the present invention, the volume ratio of the beef extract culture medium and 3- Cyclohexylamino -1- propane sulfonic acid buffer solutions is preferably
(70~95):(10~20), more preferably (80~90):(12~18), most preferably 85:15.The present invention is by beef extract culture
Base and 3- Cyclohexylamino -1- propane sulfonic acid buffer solution aseptically mix according to above-mentioned volume ratio, obtain pH value as 9.5~10
Microbiological culture media.The pH that the present invention first adjusts 3- Cyclohexylamino -1- propane sulfonic acid buffer solutions mixes with beef extract culture medium again
Purpose be avoid sodium hydroxide solution strong basicity destroy beef extract culture medium in nutritional ingredient.
False bacillus firmus DSM8715 is inoculated in microbiological culture media by the present invention to be cultivated.The present invention is to the vacation
Bacillus firmus DSM8715 inoculum concentration does not have particular/special requirement, specially with oese according to the conventional behaviour of the art
Make a little colony inoculation of picking in microbiological culture media.
In the present invention, the culture is preferably carried out under oscillating condition, and the frequency of the vibration is preferably 120~
180rpm, more preferably 130~160rpm, most preferably 150rpm.In the present invention, the temperature of the culture be preferably 25~
35 DEG C, more preferably 28~32 DEG C, most preferably 29~31 DEG C.In the present invention, the time of the culture is preferably 8~48h,
More preferably 10~24h, most preferably 12~20h.The present invention is not particularly limited to the instrument of the shaken cultivation, using this
The equipment that art personnel routinely select, such as constant-temperature table.
In the present invention, for the false bacillus firmus DSM8715 after microbiological culture media culture, centrifugation obtains thalline
Precipitation；In the present invention, the rotating speed of the centrifugation is preferably 5000~7000rpm, more preferably 6000rpm；The centrifugation
Time is 8~12min, more preferably 10min.After centrifugation, the bacterial sediment is washed 2~3 times with the Mineralized Culture liquid.This
The purpose that invention is precipitated using Mineralized Culture liquid washing thalline is the further beef extract and albumen removed in microbiological culture media
Peptone composition, during avoiding follow-up Mineralized Culture, the composition such as beef extract, peptone is attached to regeneration aggregate surface, influences
Reinforcing of the microorganism to regeneration aggregate.
The present invention mixes false bacillus firmus DSM8715 with Mineralized Culture liquid, obtains mixed-culture medium.In the present invention
In, the Mineralized Culture liquid preferably includes 10~45g/L 3- Cyclohexylamino -1- propane sulfonic acid, 4~12g/L using water as solvent
Sodium lactate, 1~3g/L sodium nitrate, 20~40mmol/L calcium chloride, 0.5~1.5mmol/L magnesium sulfate, 0.08~
3- Cyclohexylamino -1- propane sulfonic acid, the 6~10g/L lactic acid of 0.18mmol/L potassium dihydrogen phosphate, more preferably 15~35g/L
Sodium, 1.5~2g/L sodium nitrate, 22~30mmol/L calcium chloride, 0.75~1mmol/L magnesium sulfate, 0.1~0.13mmol/L
Potassium dihydrogen phosphate, most preferably 22.13g/L 3- Cyclohexylamino -1- propane sulfonic acid, 8.5g/L sodium lactate, 1.7g/L nitric acid
Sodium, 25.5mmol/L calcium chloride, 0.85mmol/L magnesium sulfate, 0.11mmol/L potassium dihydrogen phosphate.In the present invention, institute
3- Cyclohexylamino -1- propane sulfonic acid is stated as composition is buffered, maintains the pH stable of the Mineralized Culture liquid；The sodium lactate is micro-
Biology provides carbon source, metabolism generation carbonate or carbon dioxide after microorganism absorbs, reacts generation calcium carbonate with calcium ion；Institute
State sodium nitrate and provide nitrogen source for microorganism, for bacterium synthetic protein, nucleic acid and other nitrogen compounds；The calcium chloride is
Mineralising deposition provides calcium ion, carbonate or carbon dioxide reaction generation calcium carbonate with microbial metabolism generation；The sulfuric acid
Magnesium and potassium dihydrogen phosphate provide inorganic salts ingredients for microorganism, in the growth course of microorganism, form thalline Cell Component,
The activator or inhibitor of part, enzyme as enzyme, adjust osmotic pressure, PH, oxidation-reduction potential of culture medium etc..
In the present invention, the Mineralized Culture liquid is preferably prepared using two-step method.First making lactic acid sodium, sodium nitrate,
The mixed solution (A liquid) of calcium chloride, magnesium sulfate and potassium dihydrogen phosphate, then the water by the A liquid and 3- Cyclohexylamino -1- propane sulfonic acid
Solution (B liquid) is mixed, and obtains Mineralized Culture liquid.The pH value of A liquid or B liquid of the present invention is preferably 10.1~10.9, more
Preferably 10.4~10.6, most preferably 10.5.The present invention individually adjusts the pH value of A liquid and B liquid in process for preparation；This hair
The bright pH value that A liquid and B liquid are preferably adjusted with NaOH.Being adjusted according to NaOH solution, the concentration of the NaOH solution is preferably 1~
10mol/l, more preferably 5~7mol/l.After A liquid and B liquid are mixed, Mineralized Culture liquid is obtained；The pH of the Mineralized Culture liquid
Value is preferably 10.1~10.9, more preferably 10.4~10.6, most preferably 10.5.The present invention prepares mineralising training using two-step method
Nutrient solution, the pH of A liquid and B liquid is adjusted respectively, it is easier to control ph, so as to provide suitable pH for bacterial metabolism.Ore deposit of the present invention
When the pH value for changing nutrient solution is 10.5, bacterial growth is not only improved, is advantageous to synthetic calcium carbonate again.
The present invention mixes false bacillus firmus DSM8715 with Mineralized Culture liquid, obtains mixed-culture medium.Described mixed
Close in nutrient solution, false bacillus firmus DSM8715 bacterium number is preferably 1 × 107~109Cfu/L, more preferably 1 ×
108cfu/L。
After obtaining mixed-culture medium, the present invention soaks regeneration aggregate in the mixed-culture medium, makes false strong gemma
Bacillus DSM8715 is attached in the surface pore of regeneration aggregate.In the present invention, the granularity of the regeneration aggregate is preferably 0.10
~25mm.In an embodiment of the present invention, the granularity of regeneration aggregate is 0.15~4.75mm or 5~10mm or 10~20mm.
The present invention does not have special limitation to the source of the regeneration aggregate, using technology well known to those skilled in the art
Scheme is prepared by discarded concrete.Specifically, in an embodiment of the present invention, the preparation method of the regeneration aggregate is preferred
Comprise the following steps：
Discarded concrete is crushed, the reinforcing bar in the concrete after crushing is taken out, obtains mass concrete；By the nothing
Armored concrete crushes again, obtains 0.10~25mm regeneration aggregate.
In the present invention, the broken degree of the mass concrete is for 5~20mm or less than 4.75mm；It is described discarded mixed
The broken mode of solidifying soil is preferably to be broken off reinforcing bar or to take out reinforcing bar after hand breaking, obtain no-reinforcing-bar using quartering hammer
Concrete.The present invention is not particularly limited to the mode that mass concrete crushes again, using the conventional method in this area
.
As one of optional technical scheme, in the present invention, the quality of the regeneration aggregate and the body of mixed-culture medium
Product is than being preferably (0.5~1.5) g:(15~25) ml, more preferably (0.8~1.2) g:(18~22) ml, most preferably 1g:
20ml.Now, the time of the immersion is preferably 15~30 days, more preferably 18~25 days, most preferably 20 days.In the present invention
In, when regeneration aggregate soaks in mixed-culture medium, microbes are by being metabolized the calcium ion Ca in nutrient solution2+It is converted into carbon
Sour calcium CaCO3It is deposited in regeneration aggregate surface pore, so as to block regeneration aggregate surface pore, reduces the water suction of regeneration aggregate
Rate.In the present invention, the Mineralized Culture liquid in immersion process act as provide calcium source (calcium ion) and microorganism it is suitable
Living environment and higher mineralization activity.
As the two of optional technical scheme, in step 2), the soak time is 1~5h, more preferably 1.5~3h,
Most preferably 2h.Regeneration aggregate is soaked in mixed-culture medium, and microbes are attached in regeneration aggregate surface pore, while micro-
Biology can be by being metabolized the calcium ion Ca in nutrient solution2+It is converted into calcium carbonate CaCO3It is deposited in regeneration aggregate surface pore,
So as to block regeneration aggregate surface pore, the water absorption rate of regeneration aggregate is reduced.
After immersion terminates, separation of solid and liquid obtains carrying the regeneration aggregate of strain.The present invention sprays the Mineralized Culture liquid
Carried described in pouring on the regeneration aggregate of strain, strengthened regeneration aggregate.In the present invention, it is described to spray to keep regenerating
The surface wettability of aggregate is standard, it is preferred to use the mode of intermittent shower, spray 0.5~1.5h every 10~12h, per 1kg again
The amount of raw aggregate spray mixed-culture medium is 1~3mL/min；It is furthermore preferred that 1h is sprayed every 11h, per 1kg regeneration aggregates spray
The amount of mixed-culture medium is 2mL/min；.During spray, keep Mineralized Culture liquid temperature be 25~35 DEG C, more preferably 28
~32 DEG C, most preferably 29~31 DEG C.
The total time of the spray is 15~30d, more preferably 18~25d, most preferably 20d.Wherein, spray it is total when
Between include spray time and spray off time.In the present invention, the purpose for spraying Mineralized Culture liquid is to keep regeneration aggregate
Moistening, provide moisture and nutriment for the vital movement of microorganism.Meanwhile microorganism is by being metabolized the calcium in nutrient solution
Ion Ca2+It is converted into calcium carbonate CaCO3It is deposited in regeneration aggregate surface pore, so as to block regeneration aggregate surface pore, drop
The water absorption rate of low regeneration aggregate.In the present invention, the Mineralized Culture liquid in immersion process act as provide calcium source (calcium from
Son) and the suitable living environment of microorganism and higher mineralization activity.
After the regeneration aggregate that strengthened, preferred pair regeneration aggregate is dried the present invention.The present invention is to the drying
Method is not particularly limited, the technical scheme dried using aggregate well known to those skilled in the art；In the reality of the present invention
Apply in example, can be specifically dried using air dry oven or naturally dry.When the drying is preferably forced air drying, institute
State to dry and be preferably specially：Dried under the conditions of 25~35 DEG C to constant weight.
In the present invention, according to standard GB/T25176-2010《Concrete and mortar regeneration aggregate》At test microbes
The water absorption rate change of intensifying regenerating aggregate before and after reason.
A kind of microbial augmentation regeneration aggregate provided by the invention and preparation method thereof is carried out with reference to embodiment detailed
Thin explanation, but they can not be interpreted as limiting the scope of the present invention.
Embodiment 1
The preparation of regeneration aggregate：Discarded concrete tentatively crush with quartering hammer, crushed reinforcing bar using quartering hammer
Out, the difficult reinforcing bar taken out after hand breaking by taking out, the mass concrete tentatively crushed, then is entered with jaw crusher
Row is broken, screens out 5~10mm aggregate.
The preparation of Mineralized Culture liquid：By 8.5g sodium lactates, 1.7g sodium nitrate, 25.5mmol calcium chloride, 0.85mmol sulfuric acid
Magnesium and 0.11mmol potassium dihydrogen phosphates are dissolved in 800ml deionized waters, after fully mixing, are settled to 850mL, are obtained A liquid；Weigh
3- Cyclohexylamino -1- propane sulfonic acid 22.3g, measure deionized water 130mL, and 6mol/L sodium hydroxide and 90% is used after fully mixing
Acetum adjust pH value to 10.5, be settled to 150mL, obtain B liquid；A liquid is well mixed with B liquid, as Mineralized Culture
Liquid.
The preparation of microbiological culture media：Beef extract 3g, peptone 10g are weighed, deionized water 850mL is measured, is configured to ox
Meat extract culture medium, 121 DEG C of sterilizing 15min, is cooled to room temperature；3- Cyclohexylamino -1- propane sulfonic acid 22.3g are weighed, measure deionization
Water 130mL, pH value is adjusted to 10 with 6mol/L sodium hydroxides after fully mixing, 150mL is settled to, prepares 3- Cyclohexylaminos -1-
Propane sulfonic acid buffer solution, 121 DEG C of sterilizing 15min, is cooled to room temperature；It will be cooled to the beef extract culture medium and 3- hexamethylene ammonia of room temperature
Base -1- propane sulfonic acid buffer solution mixes, and obtains microbiological culture media.
False bacillus firmus DSM8715 is inoculated in microbiological culture media, the shaken cultivation 24h under the conditions of 30 DEG C,
The frequency of vibration is 150rpm, and culture is centrifuged after terminating and washed 3 times with Mineralized Culture liquid, obtains bacterial precipitation, bacterium is sunk
Shallow lake is added in Mineralized Culture liquid, obtains mixed-culture medium, and it is 1 × 10 to make the bacterium number in mixed-culture medium8cfu/L；Weigh again
Raw aggregate is added in mixed-culture medium, and the mass volume ratio of regeneration aggregate and mixed-culture medium is 1g:20ml, in normal temperature condition
Lower immersion 20 days, strengthened regeneration aggregate.According to standard GB/T25176-2010《Concrete and mortar regeneration aggregate》Survey
The performance such as water absorption rate of intensifying regenerating aggregate, the results are shown in Table 1 before and after examination microbiological treatment：
Intensifying regenerating aggregate performance comparison before and after the processing in the embodiment of the present invention of table 1
It can be drawn by table 1, water absorption rate of the intensifying regenerating aggregate after microbiological treatment is 7.3%, the water suction of before processing
Rate is 8.6%, and compared with before microbiological treatment, water absorption rate reduces 15.1%.
Embodiment 2
The preparation of regeneration aggregate：Discarded concrete tentatively crush with quartering hammer, crushed reinforcing bar using quartering hammer
Out, the difficult reinforcing bar taken out after hand breaking by taking out, the mass concrete tentatively crushed, then is entered with jaw crusher
Row is broken, screens out the aggregate less than 4.75mm, finally prepares 0.15~4.75mm aggregate.
The preparation of Mineralized Culture liquid：By 8.5g sodium lactates, 1.7g sodium nitrate, 25.5mmol calcium chloride, 0.85mmol sulfuric acid
Magnesium and 0.11mmol potassium dihydrogen phosphates are dissolved in 800ml deionized waters, after fully mixing, are settled to 850mL, are obtained A liquid；Weigh
3- Cyclohexylamino -1- propane sulfonic acid 22.3g, measure deionized water 130mL, with 6mol/L sodium hydroxides and 90% after fully mixing
Acetum adjusts pH value to 10.5, is settled to 150mL, obtains B liquid；A liquid is well mixed with B liquid, as Mineralized Culture liquid.
The preparation of microbiological culture media：Beef extract 3g, peptone 10g are weighed, deionized water 850mL is measured, is configured to ox
Meat extract culture medium, 121 DEG C of sterilizing 15min, is cooled to room temperature；3- Cyclohexylamino -1- propane sulfonic acid 22.3g are weighed, measure deionization
Water 130mL, pH value is adjusted to 10 with 6mol/L sodium hydroxides after fully mixing, 150mL is settled to, prepares 3- Cyclohexylaminos -1-
Propane sulfonic acid buffer solution, 121 DEG C of sterilizing 15min, is cooled to room temperature；It will be cooled to the beef extract culture medium and 3- hexamethylene ammonia of room temperature
Base -1- propane sulfonic acid buffer solution mixes, and obtains microbiological culture media.
False bacillus firmus DSM8715 is inoculated in microbiological culture media, the shaken cultivation 24h under the conditions of 30 DEG C,
The frequency of vibration is 150rpm, and culture is centrifuged after terminating and washed 3 times with Mineralized Culture liquid, obtains bacterial precipitation, bacterium is sunk
Shallow lake is added in Mineralized Culture liquid, obtains mixed-culture medium, and it is 1 × 10 to make the bacterium number in mixed-culture medium8cfu/L；Weigh again
Raw aggregate is added in mixed-culture medium, and the mass volume ratio of regeneration aggregate and mixed-culture medium is 1g:20ml, in normal temperature condition
Lower immersion 20 days, strengthened regeneration aggregate.According to standard GB/T25176-2010《Concrete and mortar regeneration aggregate》Survey
The performance such as water absorption rate of intensifying regenerating aggregate, the results are shown in Table 2~3 before and after examination microbiological treatment.
The granularity of intensifying regenerating aggregate in the embodiment of the present invention 2 of table 2
Intensifying regenerating aggregate is through the performance before and after microbiological treatment in the embodiment of the present invention 2 of table 3
It can be drawn by table 3, the water absorption rate of the intensifying regenerating aggregate after microbiological treatment is 4.9%, the suction of before processing
Water rate is 6.0%, and compared with before microbiological treatment, water absorption rate reduces 18.3%.
Embodiment 3
Regeneration mortar is prepared as reclaimed sand by the use of untreated regeneration aggregate in embodiment 2 and the regeneration aggregate after reinforcing,
Proportioning is shown in Table 4, investigates influence of the regeneration aggregate after microbial augmentation to mortar compression strength, the results are shown in Table 5.
Table 4 regenerates mortar mix ratio
Table 5 regenerates mortar compression strength
It can be drawn by table 5, after microbiological treatment, the compression strength for regenerating mortar adds 7.2~18.9%.
Embodiment 4
Regeneration mortar is prepared as reclaimed sand by the use of untreated regeneration aggregate in embodiment 2 and the regeneration aggregate after reinforcing,
Proportioning is shown in Table 6, investigates influence of the regeneration aggregate after microbial augmentation to mortar compression strength, the results are shown in Table 7.
Table 6 regenerates mortar mix ratio
Table 7 regenerates mortar compression strength/MPa
It can be drawn by table 7, after microbiological treatment, the compression strength for regenerating mortar adds 6.9~23.4%.
Embodiment 5
The preparation of regeneration aggregate：Discarded concrete tentatively crush with quartering hammer, crushed reinforcing bar using quartering hammer
Out, the difficult reinforcing bar taken out after hand breaking by taking out, the mass concrete tentatively crushed, then is entered with jaw crusher
Row is broken, screens out 5~10mm aggregate.
The preparation of Mineralized Culture liquid：By 8.5g sodium lactates, 1.7g sodium nitrate, 25.5mmol calcium chloride, 0.85mmol sulfuric acid
Magnesium and 0.11mmol potassium dihydrogen phosphates are dissolved in 800ml deionized waters, after fully mixing, are settled to 850mL, are obtained A liquid；Weigh
3- Cyclohexylamino -1- propane sulfonic acid 22.3g, measure deionized water 130mL, with 6mol/L sodium hydroxides and 90% after fully mixing
Acetum adjusts pH value to 10.5, is settled to 150mL, obtains B liquid；A liquid is well mixed with B liquid, as Mineralized Culture liquid.
The preparation of microbiological culture media：Beef extract 3g, peptone 10g are weighed, deionized water 850mL is measured, is configured to ox
Meat extract culture medium, 121 DEG C of sterilizing 15min, is cooled to room temperature；3- Cyclohexylamino -1- propane sulfonic acid 22.3g are weighed, measure deionization
Water 130mL, pH value is adjusted to 10 with 6mol/L sodium hydroxides after fully mixing, 150mL is settled to, prepares 3- Cyclohexylaminos -1-
Propane sulfonic acid buffer solution, 121 DEG C of sterilizing 15min, is cooled to room temperature；It will be cooled to the beef extract culture medium and 3- hexamethylene ammonia of room temperature
Base -1- propane sulfonic acid buffer solution mixes, and obtains microbiological culture media.
False bacillus firmus DSM8715 is inoculated in microbiological culture media, the shaken cultivation 24h under the conditions of 30 DEG C,
The frequency of vibration is 150rpm, and culture is centrifuged after terminating and washed 3 times with Mineralized Culture liquid, obtains bacterial precipitation, bacterium is sunk
Shallow lake is added in Mineralized Culture liquid, obtains mixed-culture medium, and it is 1 × 10 to make the bacterium number in mixed-culture medium8cfu/L.Will regeneration
Aggregate, which is added in mixed-culture medium, soaks 2h, separation of solid and liquid, obtains carrying the regeneration aggregate of strain.Then by Mineralized Culture
Liquid intermittent shower sprays 1h on the regeneration aggregate for carrying strain every 11h, per the spray mixing training of 1kg regeneration aggregates
The amount of nutrient solution is 2mL/min；.After intermittent shower 20d, strengthened regeneration aggregate.According to standard GB/T25176-2010《Coagulation
Soil and mortar regeneration aggregate》The test microbes performance such as water absorption rate of intensifying regenerating aggregate before and after the processing, the results are shown in Table 8.
Intensifying regenerating aggregate performance comparison before and after the processing in the embodiment of the present invention of table 8
It can be drawn by table 8, water absorption rate of the intensifying regenerating aggregate after microbiological treatment is 6.9%, the water suction of before processing
Rate is 8.6%, and compared with before microbiological treatment, water absorption rate reduces 19.8%.
Embodiment 6
The preparation of regeneration aggregate：Discarded concrete tentatively crush with quartering hammer, crushed reinforcing bar using quartering hammer
Out, the difficult reinforcing bar taken out after hand breaking by taking out, the mass concrete tentatively crushed, then is entered with jaw crusher
Row is broken, screens out 0.15~4.75mm aggregate.
The preparation of Mineralized Culture liquid：By 8.5g sodium lactates, 1.7g sodium nitrate, 25.5mmol calcium chloride, 0.85mmol sulfuric acid
Magnesium and 0.11mmol potassium dihydrogen phosphates are dissolved in 800ml deionized waters, after fully mixing, are settled to 850mL, are obtained A liquid；Weigh
3- Cyclohexylamino -1- propane sulfonic acid 22.3g, measure deionized water 130mL, with 6mol/L sodium hydroxides and 90% after fully mixing
Acetum adjusts pH value to 10.5, is settled to 150mL, obtains B liquid；A liquid is well mixed with B liquid, as Mineralized Culture liquid.
The preparation of microbiological culture media：Beef extract 3g, peptone 10g are weighed, deionized water 850mL is measured, is configured to ox
Meat extract culture medium, 121 DEG C of sterilizing 15min, is cooled to room temperature；3- Cyclohexylamino -1- propane sulfonic acid 22.3g are weighed, measure deionization
Water 130mL, pH value is adjusted to 10 with 6mol/L sodium hydroxides after fully mixing, 150mL is settled to, prepares 3- Cyclohexylaminos -1-
Propane sulfonic acid buffer solution, 121 DEG C of sterilizing 15min, is cooled to room temperature；It will be cooled to the beef extract culture medium and 3- hexamethylene ammonia of room temperature
Base -1- propane sulfonic acid buffer solution mixes, and obtains microbiological culture media.
False bacillus firmus DSM8715 is inoculated in microbiological culture media, the shaken cultivation 24h under the conditions of 30 DEG C,
The frequency of vibration is 150rpm, and culture is centrifuged after terminating and washed 3 times with Mineralized Culture liquid, obtains bacterial precipitation, bacterium is sunk
Shallow lake is added in Mineralized Culture liquid, obtains mixed-culture medium, and it is 1 × 10 to make the bacterium number in mixed-culture medium8cfu/L.Will regeneration
Aggregate, which is added in mixed-culture medium, soaks 2h, separation of solid and liquid, obtains carrying the regeneration aggregate of strain.Then by Mineralized Culture
Liquid intermittent shower sprays 1h on the regeneration aggregate for carrying strain every 11h, per the spray mixing training of 1kg regeneration aggregates
Support
The amount of liquid is 2mL/min；.After intermittent shower 20d, strengthened regeneration aggregate.According to standard GB/T25176-
2010《Concrete and mortar regeneration aggregate》The test microbes performance such as water absorption rate of intensifying regenerating aggregate before and after the processing, as a result
It is shown in Table 9~10.
Table 9：The granularity of intensifying regenerating aggregate in the embodiment of the present invention 6
Intensifying regenerating aggregate is through the performance before and after microbiological treatment in the embodiment of the present invention 6 of table 10
It can be drawn by table 10, the water absorption rate of the intensifying regenerating aggregate after microbiological treatment is 4.3%, the suction of before processing
Water rate is 6.0%, and compared with before microbiological treatment, water absorption rate reduces 28.3%.
Embodiment 7
Regeneration mortar is prepared as reclaimed sand by the use of untreated regeneration aggregate in embodiment 6 and the regeneration aggregate after reinforcing,
Proportioning is shown in Table 11, investigates influence of the regeneration aggregate after microbial augmentation to mortar compression strength, the results are shown in Table 12.
Table 11 regenerates mortar mix ratio
Table 12 regenerates mortar compression strength/MPa
It can be drawn by table 12, after microbiological treatment, the compression strength of regeneration mortar prepared by regeneration aggregate significantly carries
It is high.
Embodiment 8
Regeneration mortar is prepared as reclaimed sand by the use of untreated regeneration aggregate in embodiment 6 and the regeneration aggregate after reinforcing,
Proportioning is shown in Table 13, investigates influence of the regeneration aggregate after microbial augmentation to mortar compression strength, the results are shown in Table 14.
Table 13 regenerates mortar mix ratio
Table 14 regenerates mortar compression strength
It can be drawn by table 14, after microbiological treatment, the compression strength of regeneration mortar prepared by regeneration aggregate significantly carries
It is high.
As seen from the above embodiment, concrete regenerating is improved using false bacillus firmus DSM8715 provided by the invention
The method of aggregate, the water absorption rate and crush index value of obtained regeneration aggregate reduce, after preparing regeneration mortar by regeneration aggregate, then
The compression strength increase of greensand slurry.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of method that false bacillus firmus DSM8715 improves regenerative bone material by concrete, comprises the following steps：
1) false bacillus firmus DSM8715 is mixed with Mineralized Culture liquid, obtains mixed-culture medium；The Mineralized Culture liquid
3- Cyclohexylamino -1- propane sulfonic acid of the preparation raw material including 10~45g/L, 4~12g/L sodium lactate, 1~3g/L sodium nitrate, 20
~40mmol/L calcium chloride, 0.5~1.5mmol/L magnesium sulfate, 0.08~0.18mmol/L potassium dihydrogen phosphate；
2) regeneration aggregate is soaked in the mixed-culture medium that the step 1) obtains, strengthened regeneration aggregate.
2. false bacillus firmus DSM8715 improves the method for regenerative bone material by concrete according to claim 1, its feature exists
In the pH value of the Mineralized Culture liquid is 10.1~10.9.
3. false bacillus firmus DSM8715 improves the method for regenerative bone material by concrete according to claim 1, its feature exists
In in the mixed-culture medium that the step 1) obtains, false bacillus firmus DSM8715 bacterium number is 1 × 107~109cfu/L。
4. false bacillus firmus DSM8715 improves the method for regenerative bone material by concrete according to claim 1, its feature exists
In step 1) the false bacillus firmus DSM8715 is obtained through microbiological culture media culture；
The microbiological culture media includes beef extract culture medium and 3- Cyclohexylamino -1- propane sulfonic acid buffer solutions；The beef extract training
Base is supported using water as solvent, including 2.5~4.5g/L beef extract and 8~15g/L peptone；3- Cyclohexylaminos-the 1- third
Sulfonate buffer includes 120~170g/L 3- Cyclohexylamino -1- propane sulfonic acid using water as solvent；3- Cyclohexylaminos-the 1- third
The pH value of sulfonate buffer is 9~10.5；The volume ratio of the beef extract culture medium and 3- Cyclohexylamino -1- propane sulfonic acid buffer solutions
For (70~95):(10~20).
5. false bacillus firmus DSM8715 improves the method for regenerative bone material by concrete according to claim 4, its feature exists
In the culture is carried out under oscillating condition, and the frequency of the vibration is 120~180rpm；The temperature of the culture be 25~
35 DEG C, the time of the culture is 8~48h.
6. false bacillus firmus DSM8715 improves the method for regenerative bone material by concrete according to claim 4, its feature exists
In for the false bacillus firmus DSM8715 after microbiological culture media culture, centrifugation obtains bacterial sediment；With the mineralising
The nutrient solution washing bacterial sediment 2~3 times；The rotating speed of the centrifugation is 5000~7000rpm, and the time of the centrifugation is 8
~12 minutes.
7. false bacillus firmus DSM8715 improves the side of regenerative bone material by concrete according to claim 1~6 any one
Method, it is characterised in that the quality of the regeneration aggregate in the step 2) and the volume ratio of mixed-culture medium are (0.5~1.5) g:
(15~25) ml.
8. false bacillus firmus DSM8715 improves the method for regenerative bone material by concrete according to claim 7, its feature exists
In the time of the step 2) immersion is 15~30 days.
9. false bacillus firmus DSM8715 improves the side of regenerative bone material by concrete according to claim 1~6 any one
Method, it is characterised in that in step 2), the regeneration aggregate also includes after being soaked in mixed-culture medium：With ore deposit step 1) described
Change nutrient solution and spray the regeneration aggregate after the immersion, strengthened regeneration aggregate；The time of the immersion is 1~5h.
10. false bacillus firmus DSM8715 improves the method for regenerative bone material by concrete, its feature according to claim 9
It is, the mode of the spray is intermittent shower, and 0.5~1.5h is sprayed every 10~12h, per the spray mixing of 1kg regeneration aggregates
The amount of nutrient solution is 1~3mL/min；The total time of the spray is 15~30d.
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