CN112812333A - Preparation method and application of perishable garbage fermentation bacterium carrier - Google Patents
Preparation method and application of perishable garbage fermentation bacterium carrier Download PDFInfo
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Abstract
The invention relates to the technical field of fermentation inoculum carriers, in particular to a preparation method and application of a perishable garbage fermentation inoculum carrier, wherein the method comprises the following steps: preparing sodium alginate and polyvinyl alcohol aqueous solution, adding grafted cellulose nanocrystalline and iron sugar modified biochar, crosslinking by calcium chloride solution, then placing in solution containing sulfuric acid and formaldehyde, stirring for reaction, washing with clear water to be neutral, and drying to obtain the sodium alginate/polyvinyl alcohol aqueous solution; the use comprises immobilizing perishable waste ferments with the carrier. The preparation method solves the problems that the traditional carrier is low in strength, weak in microorganism fixing effect and incapable of being reused, provides the preparation method of the perishable garbage fermentation inoculant carrier with high adsorption rate and mechanical strength, and the carrier prepared by the preparation method can be reused.
Description
Technical Field
The invention relates to the technical field of fermentation inoculant carriers, in particular to a preparation method and application of a perishable garbage fermentation inoculant carrier.
Background
The immobilized microorganism technology is applied to degradation treatment of wastewater, kitchen waste and the like more and more in the year, and particularly is a biological technology which fixes microorganisms on a microbial inoculum carrier, enables the microorganisms to be highly dense and keeps biological activity and can quickly proliferate in large quantities under appropriate conditions. Generally, for a specific object to be treated, microorganisms from a natural environment often have the defects of difficult preservation, difficult enrichment, faster consumption, lower efficiency, difficult control and the like, so that after a microbial preparation with known degradation capability is fixed to a specific carrier through an immobilization technology, the microbial preparation has obvious advantages in storage, circulation, treatment effect and the like.
The invention discloses a Chinese patent with an authorization publication number of CN105665417B in the prior art, and discloses a preparation method and application of a composite microbial agent for efficiently degrading kitchen waste, wherein the composite microbial agent consists of a composite thallus and a carrier, the composite thallus is formed by mixing issatchenkia orientalis, bacillus subtilis, abnormal Wilkholdham yeast, trichoderma and actinomycetes, the carrier consists of bean pulp, bran, rice hull powder and wood shavings, and the composite thallus accounts for 6-12% of the composite microbial agent by weight. The compound bacteria are fully mixed with the kitchen organic waste in fermentation equipment, and under the aerobic condition at normal temperature, different strains act synergistically, so that the degradation rate of the kitchen organic waste is improved, the fermentation time is shortened, and meanwhile, the deodorization effect is obvious; the kitchen waste fermented by the composite bacteria can be used as an organic fertilizer. Although the composite microbial agent in the method can improve the degradation rate of the organic kitchen waste and shorten the fermentation time, the carrier consisting of the bean pulp, the bran, the rice hull powder and the wood shavings is simply and physically mixed with the composite bacteria, the microorganism is adsorbed only by the polar groups on the surface of the cellulose, the carrier has a weak fixing effect on the bacteria, the carrier cannot be reused, the calculation of the verified reduction rate of the kitchen waste is wrong, and the actual degradation efficiency is low.
The above background disclosure is only for the purpose of assisting understanding of the inventive concept and technical solutions of the present invention, and does not necessarily belong to the prior art of the present patent application, and should not be used for evaluating the novelty and inventive step of the present application in the case that there is no clear evidence that the above content is disclosed at the filing date of the present patent application.
Disclosure of Invention
Technical problem to be solved
The technical problem to be solved by the invention is as follows: the preparation method solves the problems that the traditional carrier is low in strength, weak in microorganism fixing effect and incapable of being reused, provides the preparation method of the perishable garbage fermentation inoculant carrier with high adsorption rate and mechanical strength, and the carrier prepared by the preparation method can be reused.
(II) technical scheme
In order to solve the above technical problems or to achieve the above technical object, the present invention provides the following technical solutions.
The preparation method of the perishable garbage fermentation inoculant carrier comprises the following steps: preparing sodium alginate and polyvinyl alcohol aqueous solution, adding grafted cellulose nanocrystalline and iron sugar modified biochar, crosslinking by calcium chloride solution, then placing in solution containing sulfuric acid and formaldehyde, stirring for reaction, washing with clear water to neutrality, and drying to obtain the product.
Preferably, the preparation method of the perishable garbage fermentation inoculant carrier specifically comprises the following steps:
s1, adding 1 part by weight of sodium alginate and 5.5-6.2 parts by weight of polyvinyl alcohol into 200 parts by weight of deionized water, heating to 95-99 ℃ for dissolution, and naturally cooling to room temperature;
s2, 0.2-0.3 part by weight of grafted cellulose nanocrystalline and 0.5-0.6 part by weight of iron sugar modified biochar are added into the solution obtained in the step S1, and ultrasonic dispersion is carried out for at least 30 min;
s3, dripping the mixed solution obtained in the step S2 into 1-2% calcium chloride solution within 30 min-1 h for reaction for at least 12 h;
s4, filtering the formed sample, soaking the sample in a mixed solution containing 0.5-2% of sulfuric acid and 3-10% of formaldehyde, reacting for 15-45 min under slow stirring, taking out, and washing with clear water to be neutral to obtain the product.
Preferably, in step S1, the viscosity of sodium alginate is 100-400 cps.
Preferably, in step S1, the polyvinyl alcohol has a weight average molecular weight of 16000 to 24000.
Preferably, in step S1, stirring at 120-600 r/min is used to assist dissolution.
Preferably, in step S2, the weight ratio of the grafted cellulose nanocrystal to the iron sugar modified biochar is 1: 2-2.5.
Preferably, in step S2, the grafted cellulose nanocrystal is obtained by modifying the cellulose nanocrystal with carboxyl and then grafting polyethylene glycol.
Preferably, in step S2, the method for preparing the grafted cellulose nanocrystal specifically includes:
1) uniformly dissolving 1 part by weight of sodium bromide and 0.1 part by weight of 2,2,6, 6-tetramethylpiperidine-nitrogen-oxide in 100 parts by weight of distilled water, adding 1000 parts by weight of 1-1.5% by weight of cellulose nanocrystal suspension, reacting at 150-450 r/min for 15-30 min, adjusting the pH of the solution to 10.0-10.5 by using a sodium hydroxide solution, slowly dropwise adding 20-50 parts by weight of a 15% sodium hypochlorite solution, reacting for 30-60 min, adding absolute ethyl alcohol to stop the reaction, washing, centrifuging and drying to obtain carboxylated cellulose nanocrystals;
2) dispersing 10 parts by weight of carboxylated cellulose nanocrystals into 1000 parts by weight of N, N-dimethylformamide, adding 2.2-3.0 parts by weight of low molecular weight polyethylene glycol and 0.01-0.05 part by weight of dibutyltin dilaurate under the protection of nitrogen, slowly heating to 80-82 ℃, continuously reacting for 12-18 h, washing for 1-2 times by using deionized water and absolute ethyl alcohol respectively, centrifuging, and drying in vacuum to obtain the grafted modified cellulose nanocrystals.
Preferably, the preparation method of the grafted cellulose nanocrystal comprises the following limiting factors:
the mass fraction of the sodium hydroxide solution is 5-15%;
the speed of slowly dripping the sodium hypochlorite solution is 2-5 mL/min;
the addition amount of the absolute ethyl alcohol is 50-80 parts by weight;
the low molecular weight polyethylene glycol is at least one of PEG-300, PEG-400, PEG-600, PEG-800 or PEG-1000;
the slow heating rate is 3-5 ℃/min;
the vacuum drying means vacuum drying at 45-60 ℃ to constant weight.
The inventor finds that the grafted cellulose nanocrystalline obtained by the application is added into polyvinyl alcohol carrier microspheres to be beneficial to improving the content of carrier active groups and improving the mass transfer performance of a carrier to a greater extent, so that the load of microorganisms is facilitated, the load rate is obviously improved, the enrichment of the microorganisms is facilitated, the microorganism content in a fermentation microbial inoculum per unit weight is improved, and the fermentation degradation efficiency of perishable garbage is improved.
Preferably, in step S2, the iron sugar-modified biochar is obtained by high-pressure modifying oleaster dry branch biochar with ferrous ions and chitosan in a supercritical carbon dioxide device.
Preferably, in step S2, the method for preparing the iron sugar-modified biochar specifically includes:
dissolving 1 part by weight of chitosan in 100-150 parts by weight of 2-5% acetic acid solution, adding 1.2-2.0 parts by weight of biochar, and performing ultrasonic dispersion until complete mixing; and then dropwise adding a solution containing 0.5-1.0 part by weight of ferrous sulfate in a constant-pressure funnel within 1-2 h, transferring the solution into a supercritical carbon dioxide device, reacting for at least 2h under the conditions of 120-180 r/min, 42-45 ℃ and 12-20 MPa, quickly releasing pressure, taking out biochar, washing for at least 4 times by using absolute ethyl alcohol, and drying in vacuum to constant weight to obtain the iron sugar modified biochar.
Preferably, the preparation method of the iron sugar modified biochar comprises the following limiting factors:
the biochar is obtained by calcining 40-mesh oleaster dry branch powder in a muffle furnace at 400-600 ℃ for 2-4 hours;
the relative molecular mass of the chitosan is 200000-400000, and the deacetylation degree is not lower than 92%;
the ultrasonic frequency of ultrasonic dispersion is 30-40 KHz, and the ultrasonic density is 0.2-0.5W/cm2；
The mass fraction of the ferrous sulfate solution is 1-2%;
the temperature of vacuum drying is 60-80 ℃.
The oleaster dry branch biochar is subjected to chitosan and ferrous sulfate combined modification, more reactive groups such as hydroxyl groups can be introduced to the surface of the biochar, and ferrous ions are introduced to further enrich polar groups on the surface of the biochar, so that bonding between the biochar and each component of a carrier is facilitated, the mechanical strength and the chemical stability of the carrier are obviously improved, the adsorption rate to microorganisms is also improved to a certain extent, the improvement of the mechanical strength of the carrier enables the carriers to keep the structural integrity in the process of degrading perishable garbage without breaking, the carriers can adsorb the microorganisms again after regeneration to be applied to degradation of the perishable garbage, and the treatment cost is reduced.
Preferably, in step S3, the weight part of the calcium chloride solution is 300 to 500 parts by weight.
Preferably, in step S4, the volume of the mixed solution containing sulfuric acid and formaldehyde is 1.5 to 3 times the volume of the filtered sample.
Preferably, in step S4, the stirring speed of the slow stirring is 60 to 180 r/min.
According to the invention, on the basis of preparing a microbial agent carrier by cross-linking traditional sodium alginate and polyvinyl alcohol through a cross-linking agent, a certain amount of grafted cellulose nanocrystalline and ferric sugar modified biochar are doped to optimize and modify the microbial agent carrier, and the prepared carrier is subjected to a formal reaction, so that a spherical or ellipsoidal perishable garbage fermentation microbial agent carrier with a porous three-dimensional structure is finally obtained.
The application also provides a perishable waste fermentation inoculum carrier obtained by the method.
The application also provides application of the carrier in preparing a perishable waste fermentation inoculant, wherein the application comprises the step of fixing the perishable waste fermentation inoculant by using the carrier.
The above-described preferred conditions may be combined with each other to obtain a specific embodiment, in accordance with common knowledge in the art.
The raw materials or reagents involved in the invention are all common commercial products, and the operations involved are all routine operations in the field unless otherwise specified.
(III) advantageous effects
The technical scheme of the invention has the following advantages:
firstly, on the basis of preparing a microbial inoculum carrier by cross-linking traditional sodium alginate and polyvinyl alcohol, optimally modifying the microbial inoculum carrier by adding a certain amount of grafted cellulose nanocrystals and iron sugar modified biochar, and performing a formal reaction on the prepared carrier to finally obtain a spherical or ellipsoidal perishable garbage fermentation microbial inoculum carrier with a porous three-dimensional structure, wherein the carrier has excellent adsorption effect and mechanical strength;
secondly, grafting polyethylene glycol repeating units on the modified cellulose nanocrystal carboxyl to obtain grafted cellulose nanocrystals, wherein the grafted cellulose nanocrystals are added into polyvinyl alcohol carrier microspheres to facilitate the improvement of the content of carrier active groups and the improvement of the mass transfer performance of the carrier to a greater extent, so that the loading of microorganisms is facilitated, the loading rate is remarkably improved, the enrichment of microorganisms is facilitated, the content of microorganisms in a unit weight of fermentation inoculum is improved, and the fermentation degradation efficiency of perishable garbage is improved;
and thirdly, chitosan and ferrous sulfate combined modification are carried out on the oleaster dry branch biochar, more reactive groups such as hydroxyl groups can be introduced into the surface of the biochar, and ferrous ions are introduced to further enrich polar groups on the surface of the biochar, so that bonding between the biochar and each component of a carrier is facilitated, the mechanical strength and the chemical stability of the carrier are obviously improved, the adsorption rate to microorganisms is also improved to a certain extent, the improvement of the mechanical strength of the carrier enables the carriers to keep the structural integrity in the process of degrading perishable garbage, the carriers cannot be broken, the carriers can be adsorbed again to microorganisms after regeneration to be applied to degradation of perishable garbage, and the treatment cost is reduced.
The invention adopts the technical scheme for achieving the purpose, makes up the defects of the prior art, and has reasonable design and convenient operation.
Drawings
The foregoing and/or other objects, features, advantages and embodiments of the invention will be more readily understood from the following description taken in conjunction with the accompanying drawings in which:
FIG. 1 is a schematic view of a modification process of the graft-modified cellulose nanocrystal of the present invention;
FIG. 2 is a graph showing the breakage rate of the vectors obtained in the examples of the present invention.
Detailed Description
Those skilled in the art can appropriately substitute and/or modify the process parameters to implement the present disclosure, but it is specifically noted that all similar substitutes and/or modifications will be apparent to those skilled in the art and are deemed to be included in the present invention. While the products and methods of making described herein have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the products and methods of making described herein may be made and utilized without departing from the spirit and scope of the invention.
Unless otherwise indicated, "%" refers to weight percent and all ratios are weight ratios.
Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The present invention uses the methods and materials described herein; other suitable methods and materials known in the art may be used. The materials, methods, and examples described herein are illustrative only and are not intended to be limiting. All publications, patent applications, patents, provisional applications, database entries, and other references mentioned herein, and the like, are incorporated by reference herein in their entirety. In case of conflict, the present specification, including definitions, will control.
The materials, methods, and examples described herein are illustrative only and not intended to be limiting unless otherwise specified. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described herein.
The present invention is described in detail below.
Example 1:
the embodiment provides a perishable garbage fermentation inoculant carrier, and the specific preparation method comprises the following steps:
s1, 1g of sodium alginate (with the viscosity of 200 +/-20 cps) and 5.5g of polyvinyl alcohol (with the weight-average molecular weight of 16000) are added into 200g of deionized water, the temperature is raised to 95 ℃, the mixture is stirred at 120r/min until the mixture is dissolved, and the mixture is naturally cooled to the room temperature;
S2、
s201, uniformly dissolving 1g of sodium bromide and 0.1g of 2,2,6, 6-tetramethylpiperidine-nitrogen-oxide in 100g of distilled water, adding 1000g of cellulose nanocrystal suspension with the mass fraction of 1%, reacting at 150r/min for 30min, adjusting the pH of the solution to 10.0 by using a 5% sodium hydroxide solution, slowly dropwise adding 20g of 15% sodium hypochlorite solution at 2mL/min, reacting for 30min, adding 50g of absolute ethyl alcohol to stop the reaction, washing, centrifuging and drying to obtain carboxylated cellulose nanocrystals;
s202, dispersing 10g of carboxylated cellulose nanocrystals into 1000g N, N-dimethylformamide, adding 2.2g of PEG-400 and 0.01g of dibutyltin dilaurate under the protection of nitrogen, slowly heating to 80 ℃ at the speed of 3 ℃/min, continuously reacting for 18h, washing for 1 time by deionized water and absolute ethyl alcohol respectively, centrifuging, and drying in vacuum at the temperature of 45 ℃ until the weight is constant to obtain the grafted modified cellulose nanocrystals, wherein the preparation process is shown in figure 1;
s203, calcining the oleaster dry branch powder of 40 meshes in a muffle furnace at 400 ℃ for 4 hours to obtain biochar, dissolving 1g of chitosan (relative molecular mass is 200000, degree of deacetylation is 92%) in 100g of 2% acetic acid solution, adding 1.2g of biochar, and reacting at the frequency of 30KHz and the density of 0.2W/cm2Ultrasonic dispersion under the condition until complete mixing; then dropwise adding 25g of 2% ferrous sulfate solution within 1h by using a constant-pressure funnel, transferring the solution into a supercritical carbon dioxide device, reacting for 4h at 42 ℃ and 12MPa at 120r/min, quickly releasing pressure, taking out biochar, washing for 4 times by using absolute ethyl alcohol, and drying in vacuum at 60 ℃ to constant weight to obtain the iron sugar modified biochar;
s204, 0.2g of the grafted cellulose nanocrystalline obtained in the step S202 and 0.5g of the iron sugar modified biochar obtained in the step S203 are added into the solution obtained in the step S1, and the frequency is 50KHz, and the density is 0.6W/cm2Ultrasonic dispersing for 60min under the condition;
s3, dripping the mixed solution obtained in the step S2 into 300g of 2% calcium chloride solution within 30min to react for 24 h;
s4, filtering the formed sample, soaking the sample in a mixed solution which is 1.5 times of the volume of the sample and contains 2% of sulfuric acid and 10% of formaldehyde, slowly stirring for reaction at 60r/min for 45min, taking out the sample, and washing the sample with clear water until the sample is neutral.
Example 2:
the embodiment provides a perishable garbage fermentation inoculant carrier, and the specific preparation method comprises the following steps:
s1, 1g of sodium alginate (viscosity is 300 +/-20 cps) and 6.2g of vinyl alcohol (weight average molecular weight is 24000) are added into 200g of deionized water, the temperature is raised to 99 ℃, then 600r/min of the mixture is stirred until the mixture is dissolved, and the mixture is naturally cooled to room temperature;
S2、
s201, uniformly dissolving 1g of sodium bromide and 0.1g of 2,2,6, 6-tetramethylpiperidine-nitrogen-oxide in 100g of distilled solution, adding the distilled solution into 1000g of cellulose nanocrystal suspension with the mass fraction of 1.5%, reacting at 450r/min for 15min, adjusting the pH of the solution to 10.5 by using 15% sodium hydroxide solution, slowly dropwise adding 50g of 15% sodium hypochlorite solution at 5mL/min, reacting for 60min, adding 80g of absolute ethyl alcohol to stop the reaction, washing, centrifuging and drying to obtain carboxylated cellulose nanocrystals; dispersing 10g of carboxylated cellulose nanocrystal in 1000g N N-dimethylformamide, adding 3.0g of PEG-1000 and 0.05g of dibutyltin dilaurate under the protection of nitrogen, slowly heating to 82 ℃ at the speed of 5 ℃/min, continuously reacting for 12h, washing for 2 times by deionized water and absolute ethyl alcohol respectively, centrifuging, and drying in vacuum at the temperature of 60 ℃ until the weight is constant to obtain the graft modified cellulose nanocrystal;
s202, calcining the oleaster dry branch powder of 40 meshes in a muffle furnace at 600 ℃ for 2h to obtain biochar, dissolving 1g of chitosan (the relative molecular mass is 400000, the deacetylation degree is 94%) in 150g of 2% acetic acid solution, adding 2.0g of biochar, and reacting at the frequency of 40KHz and the density of 0.5W/cm2Ultrasonic dispersion under the condition until complete mixing; then, 100g of 1% ferrous sulfate solution is dropwise added into a constant pressure funnel within 2h, the obtained mixture is moved into a supercritical carbon dioxide device, the obtained product reacts for 2h at the temperature of 45 ℃ and 20MPa at 180r/min, the obtained product is quickly decompressed, the biochar is taken out, washed for 4 times by absolute ethyl alcohol, and dried in vacuum at the temperature of 80 ℃ to constant weight, and the iron sugar modified biochar is obtained;
s203, 0.3g of the grafted cellulose nanocrystalline obtained in the step S201 and 0.6g of the iron sugar modified biochar obtained in the step S202 are added into the solution obtained in the step S1, and the mixture is subjected to reaction at the frequency of 80KHz and the density of 1.0W/cm2Ultrasonic dispersing for 30min under the condition;
s3, dripping the mixed solution obtained in the step S2 into 500g of 1% calcium chloride solution within 1h for reaction for 12 h;
s4, filtering the formed sample, soaking the sample in a mixed solution which is 3 times of the volume of the sample and contains 0.5 percent of sulfuric acid and 3 percent of formaldehyde, slowly stirring the mixture for reaction for 15min at a speed of 180r/min, taking the mixture out, and washing the mixture with clear water to be neutral to obtain the product.
Example 3:
the embodiment provides a perishable garbage fermentation inoculant carrier, and the specific preparation method comprises the following steps:
s1, adding 1g of sodium alginate (with the viscosity of 200 +/-20 cps) and 6g of polyvinyl alcohol (with the weight-average molecular weight of 21000) into 200g of deionized water, heating to 98 ℃, stirring at 300r/min until the mixture is dissolved, and naturally cooling to room temperature;
S2、
s201, uniformly dissolving 1g of sodium bromide and 0.1g of 2,2,6, 6-tetramethylpiperidine-nitrogen-oxide in 100g of distilled water, adding 1000g of cellulose nanocrystal suspension with the mass fraction of 1.2%, reacting at 300r/min for 20min, adjusting the pH of the solution to 10.0 by using 10% sodium hydroxide solution, slowly dropwise adding 40g of 15% sodium hypochlorite solution at 5mL/min, reacting for 45min, adding 60g of absolute ethyl alcohol to stop reaction, washing, centrifuging and drying to obtain carboxylated cellulose nanocrystals; dispersing 10g of carboxylated cellulose nanocrystal in 1000g N N-dimethylformamide, adding 2.5g of PEG-600 and 0.02g of dibutyltin dilaurate under the protection of nitrogen, slowly heating to 80 ℃ at the speed of 4 ℃/min, continuously reacting for 16h, washing for 2 times by deionized water and absolute ethyl alcohol respectively, centrifuging, and drying in vacuum at the temperature of 55 ℃ until the weight is constant to obtain the graft modified cellulose nanocrystal;
s202, calcining the oleaster dry branch powder of 40 meshes in a muffle furnace at 550 ℃ for 3h to obtain biochar, dissolving 1g of chitosan (the relative molecular mass is 250000, the deacetylation degree is 92%) in 120g of 3% acetic acid solution, adding 2g of biochar, and reacting at the frequency of 35KHz and the density of 0.4W/cm2Ultrasonic dispersion under the condition until complete mixing; then, dropwise adding 80g of 1% ferrous sulfate solution within 2h by using a constant-pressure funnel, transferring the mixture into a supercritical carbon dioxide device, reacting for 3h at the temperature of 45 ℃ and 15MPa at the speed of 150r/min, quickly releasing pressure, taking out biochar, washing for 5 times by using absolute ethyl alcohol, and drying in vacuum at the temperature of 65 ℃ to constant weight to obtain the iron sugar modified biochar;
s203, 0.3g of the grafted cellulose nanocrystalline obtained in the step S201 and 0.6g of the iron sugar modified biochar obtained in the step S202 are added into the solution obtained in the step S1, and the frequency is 60KHz, and the density is 0.8W/cm2Ultrasonic dispersing for 45min under the condition;
s3, dripping the mixed solution obtained in the step S2 into 400g of 1.5% calcium chloride solution within 45min to react for 18 h;
s4, filtering the formed sample, soaking the sample in a mixed solution which is 2 times of the sample volume and contains 1% of sulfuric acid and 5% of formaldehyde, slowly stirring at 120r/min for reaction for 30min, taking out the sample, and washing the sample with clear water until the sample is neutral.
Example 4:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in this example, the cellulose nanocrystals are only carboxylated and not grafted with PEG, i.e., they are used as raw materials to participate in the reaction of step S203.
Example 5:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in this example, neither the cellulose nanocrystals were subjected to carboxylation modification nor PEG was grafted, i.e., the cellulose nanocrystals were used as the raw material to participate in the reaction of step S203, i.e., the cellulose nanocrystals were directly used to participate in the reaction of step S203.
Example 6:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in the embodiment, the corn straw powder is used for replacing the oleaster dry branch powder to prepare the biochar.
Example 7:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in this example, the oleaster biochar was not chitosan modified.
Example 8:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in this example, no ferrous sulfate modification was performed on the russianolive biochar.
Example 9:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in this example, the russianolive biochar is not modified by chitosan nor by ferrous sulfate, i.e. the rusnolive biochar is directly involved in the reaction of step S203.
Example 10:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in the present example, when the reaction of step S203 is performed, no graft-modified cellulose nanocrystal is added, and only the iron sugar-modified biochar participates in the reaction.
Example 11:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in the present example, when the reaction of step S203 is performed, no iron sugar-modified biochar is added, and only the grafted cellulose nanocrystals participate in the reaction.
Example 12:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in this example, the operation of step S2 is not performed, but the mixed solution of S1 is directly subjected to the crosslinking reaction of S3, that is, the carrier prepared in the present application does not contain any grafted cellulose nanocrystals and the iron sugar-modified biochar.
Example 13:
the example provides another perishable garbage fermentation inoculum carrier, and the specific preparation method is basically the same as that of the example 3, except that: in this example, the operation of step S4 is not performed, but the step S3 is directly reflected to that the filtered carrier is washed with clean water to be neutral, so that the perishable garbage fermentation inoculum carrier is obtained.
Experimental example 1:
this experimental example provides the detection to the mechanical strength of the perishable rubbish fermentation inoculant carrier that example 1 ~ 13 obtained, and the concrete step is: respectively placing the carriers obtained in the examples into conical flasks filled with 400mL of water samples, placing the conical flasks into a shaking table at 240r/min, stirring and oscillating the conical flasks for 48h, taking out the conical flasks, recording the damage condition of the particles, calculating the damage rate according to the ratio of the number of the damaged particles to the number of the initial particles, representing the mechanical strength according to the size of the damage rate, wherein the smaller the damage rate is, the higher the mechanical strength is, the better the structure retention capacity is, and the statistical result is shown in FIG. 2. As can be seen from FIG. 2, the perishable garbage fermentation inoculum carriers in the examples 1-3 of the preferred embodiment of the application have small damage rate which is not more than 2%, which indicates that the perishable garbage fermentation inoculum carriers have excellent mechanical strength and structure retention capacity; compared with examples 4-13, it can be found that the mechanical strength of the carrier can be weakened when the cellulose nanocrystals are not grafted and the biological carbon is not subjected to iron sugar modification, particularly, the modification of the biological carbon is particularly important, the introduction of carboxyl, ferrous ions and the like is beneficial to the bonding of the biological carbon and each component of the carrier, the mechanical strength of the biological carbon is improved, and in addition, the mechanical strength of the carrier can also be improved when the carrier is subjected to a formalization reaction.
Experimental example 2:
this experimental example provides the detection to the chemical stability of the perishable rubbish fermentation inoculant carrier that example 1 ~ 13 obtained, and the concrete step is: the carrier obtained in each example was placed in a conical flask containing 400mL of a sodium hydroxide solution with pH10.0, the dissolved amount of the particles was recorded after soaking for 12h, the deformation rate was calculated from the ratio of the number of particles with loose structure and solid dissolved out to the initial number of particles, the chemical stability was characterized by the size of the deformation rate, the smaller the deformation rate, the more excellent the chemical stability, and the statistical results are shown in Table 1.
TABLE 1 deformation ratio
| Examples of the invention | Percent deformation in sodium |
| 1 | 19.6 |
| 2 | 24.1 |
| 3 | 17.4 |
| 4 | 40.1 |
| 5 | 43.6 |
| 6 | 58.5 |
| 7 | 60.1 |
| 8 | 63.3 |
| 9 | 65.5 |
| 10 | 45.3 |
| 11 | 68.3 |
| 12 | 70.2 |
| 13 | 76.8 |
As can be seen from Table 1, the perishable garbage fermentation agent carriers in the examples 1-3 of the preferred embodiments of the present application have small deformation rates, which are not more than 25%, indicating that the carriers have excellent structure retention capacity in alkaline environment; basically similar to the verification result of the experimental example 1, the improvement of the chemical stability of the carrier is large by carrying out iron sugar modification on the biochar, the introduction of carboxyl, ferrous ions and the like is beneficial to the bonding of the biochar and each component of the carrier, the chemical structure fastness of the biochar is improved, and the chemical stability of the carrier can also be improved by carrying out a formalization reaction on the carrier.
Experimental example 3:
this experimental example provides the detection of the adsorption of microorganisms to the perishable waste fermentation inoculant carrier obtained in examples 1-13, and the specific steps are as follows:
1) preparing a bacterial liquid: activating and carrying out amplification culture on microorganisms to prepare a bacterial liquid: containing 2.5X 109cfu/mL Bacillus amyloliquefaciens, 2.5X 109cfu/mL of a composite bacterial liquid of pseudomonas azotoformans;
2) adsorption: respectively adding the carriers obtained in examples 1-13 into the composite bacterial liquid, wherein the addition amount is 300g/L, stirring for 30min at the rotating speed of 120r/min for fully mixing, standing for 12h, and separating to obtain a perishable garbage zymogen;
3) counting: diluting each bacterial liquid adsorbed by the carrier into an original volume, counting the concentration of the residual bacteria by a dilution method, and calculating the adsorption rate by combining the original concentration of the bacteria:
4) regeneration performance: and (3) after non-degradable substances in the perishable garbage are picked out, crushing and screening the perishable garbage by a 40-mesh sieve, mixing the perishable garbage with each microbial inoculum according to the weight ratio of 200:1, ventilating and air exchanging once per hour at room temperature, turning over the mixture once, degrading for 48 hours, screening out unbroken microbial inoculum, washing with deionized water, and repeating the step 2) to adsorb the microbial inoculum to obtain the regeneration adsorption rate.
The results of statistics of the adsorption rate and the regeneration adsorption rate of each carrier of examples 1 to 13 are shown in Table 2.
TABLE 2 adsorption Rate
| Examples of the invention | Adsorption rate/%) | Regeneration adsorption rate/% |
| 1 | 95.5 | 82.0 |
| 2 | 95.0 | 83.6 |
| 3 | 96.3 | 85.5 |
| 4 | 75.3 | 52.2 |
| 5 | 72.4 | 49.6 |
| 6 | 90.2 | 46.8 |
| 7 | 88.3 | 31.2 |
| 8 | 85.4 | 30.8 |
| 9 | 84.8 | 28.4 |
| 10 | 68.5 | 32.2 |
| 11 | 81.9 | 24.3 |
| 12 | 63.2 | 12.5 |
| 13 | 80.4 | 30.0 |
As can be seen from table 2 above, the carriers in preferred embodiment examples 1-3 of the present application all have an adsorption rate of not less than 95% and a regeneration adsorption rate of not less than 80% for the complex bacteria liquid, which is beneficial to the loading of microorganisms, significantly improves the loading rate, is beneficial to the enrichment of microorganisms, improves the microorganism content in the fermentation bacteria agent per unit weight, further improves the fermentation degradation efficiency of perishable garbage, and has a positive economic significance in that the treatment cost can be significantly reduced due to the higher regeneration adsorption rate. From table 2, it can be seen that whether the adsorption rate of the carrier is greatly affected by performing carboxyl modification on the cellulose nanocrystals and grafting polyethylene glycol repeating units, and the joint modification of chitosan and ferrous sulfate on the oleaster dry branch biochar is helpful to improve the maintenance of the structure of the oleaster dry branch biochar, so that the regeneration adsorption is facilitated.
Conventional techniques in the above examples are known to those skilled in the art and will not be described in detail herein.
The specific examples described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Although the present invention has been described in detail and with reference to specific examples thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.
While the above detailed description has shown, described, and pointed out novel features as applied to various embodiments, it will be understood that various omissions, substitutions, and changes in the form and details of the device or method illustrated may be made without departing from the spirit of the disclosure. In addition, the various features and methods described above may be used independently of one another, or may be combined in various ways. All possible combinations and sub-combinations are intended to fall within the scope of the present disclosure. Many of the embodiments described above include similar components, and thus, these similar components are interchangeable in different embodiments. While the invention has been disclosed in the context of certain embodiments and examples, it will be understood by those skilled in the art that the invention extends beyond the specifically disclosed embodiments to other alternative embodiments and/or uses and obvious modifications and equivalents thereof. Accordingly, the invention is not intended to be limited by the specific disclosure of preferred embodiments herein.
The invention is not the best known technology.
Claims (10)
1. The preparation method of the perishable garbage fermentation inoculant carrier is characterized by comprising the following steps: preparing sodium alginate and polyvinyl alcohol aqueous solution, adding grafted cellulose nanocrystalline and iron sugar modified biochar, crosslinking by calcium chloride solution, then placing in solution containing sulfuric acid and formaldehyde, stirring for reaction, washing with clear water to neutrality, and drying to obtain the product.
2. The method of claim 1, wherein: the preparation method of the perishable garbage fermentation inoculant carrier specifically comprises the following steps:
s1, adding 1 part by weight of sodium alginate and 5.5-6.2 parts by weight of polyvinyl alcohol into 200 parts by weight of deionized water, heating to 95-99 ℃ for dissolution, and naturally cooling to room temperature;
s2, 0.2-0.3 part by weight of grafted cellulose nanocrystalline and 0.5-0.6 part by weight of iron sugar modified biochar are added into the solution obtained in the step S1, and ultrasonic dispersion is carried out for at least 30 min;
s3, dripping the mixed solution obtained in the step S2 into 1-2% calcium chloride solution within 30 min-1 h for reaction for at least 12 h;
s4, filtering the formed sample, soaking the sample in a mixed solution containing 0.5-2% of sulfuric acid and 3-10% of formaldehyde, reacting for 15-45 min under slow stirring, taking out, and washing with clear water to be neutral to obtain the product.
3. The method according to claim 1 or 2, characterized in that:
the viscosity of the sodium alginate is 100-400 cps; and/or
The weight average molecular weight of the polyvinyl alcohol is 16000-24000.
4. The method according to claim 1 or 2, characterized in that: the weight ratio of the grafted cellulose nanocrystal to the iron sugar modified biochar is 1: 2-2.5.
5. The method according to claim 1 or 2, characterized in that: the grafted cellulose nanocrystal is obtained by grafting polyethylene glycol onto the cellulose nanocrystal after carboxyl modification.
6. The method according to claim 1 or 2, characterized in that: the preparation method of the grafted cellulose nanocrystal specifically comprises the following steps:
1) uniformly dissolving 1 part by weight of sodium bromide and 0.1 part by weight of 2,2,6, 6-tetramethylpiperidine-nitrogen-oxide in 100 parts by weight of distilled water, adding 1000 parts by weight of 1-1.5% by weight of cellulose nanocrystal suspension, reacting at 150-450 r/min for 15-30 min, adjusting the pH of the solution to 10.0-10.5 by using a sodium hydroxide solution, slowly dropwise adding 20-50 parts by weight of a 15% sodium hypochlorite solution, reacting for 30-60 min, adding absolute ethyl alcohol to stop the reaction, washing, centrifuging and drying to obtain carboxylated cellulose nanocrystals;
2) dispersing 10 parts by weight of carboxylated cellulose nanocrystals into 1000 parts by weight of N, N-dimethylformamide, adding 2.2-3.0 parts by weight of low molecular weight polyethylene glycol and 0.01-0.05 part by weight of dibutyltin dilaurate under the protection of nitrogen, slowly heating to 80-82 ℃, continuously reacting for 12-18 h, washing for 1-2 times by using deionized water and absolute ethyl alcohol respectively, centrifuging, and drying in vacuum to obtain the grafted modified cellulose nanocrystals.
7. The method according to claim 1 or 2, characterized in that: the iron sugar modified biochar is obtained by performing high-pressure modification on the oleaster dry branch biochar by utilizing ferrous ions and chitosan in a supercritical carbon dioxide device.
8. The method according to claim 1 or 2, characterized in that: the preparation method of the iron sugar modified biochar specifically comprises the following steps:
dissolving 1 part by weight of chitosan in 100-150 parts by weight of 2-5% acetic acid solution, adding 1.2-2.0 parts by weight of Elaeagnus angustifolia dry branch biochar, and performing ultrasonic dispersion until complete mixing; and then dropwise adding a solution containing 0.5-1.0 part by weight of ferrous sulfate in a constant-pressure funnel within 1-2 h, transferring the solution into a supercritical carbon dioxide device, reacting for at least 2h under the conditions of 120-180 r/min, 42-45 ℃ and 12-20 MPa, quickly releasing pressure, taking out biochar, washing for at least 4 times by using absolute ethyl alcohol, and drying in vacuum to constant weight to obtain the iron sugar modified biochar.
9. A perishable waste fermentation inoculant carrier obtainable by the method of any one of claims 1 to 8.
10. Use of a vector according to claim 9 in the preparation of a perishable waste fermentation inoculant, said use comprising immobilising perishable waste fermentation with said vector.
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