CN115125166B - Method for enhancing anaerobic treatment of kitchen waste by using conductive carrier to form Shewanella electroactive biomembrane - Google Patents
Method for enhancing anaerobic treatment of kitchen waste by using conductive carrier to form Shewanella electroactive biomembrane Download PDFInfo
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- C12N1/20—Bacteria; Culture media therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
- B09B3/60—Biochemical treatment, e.g. by using enzymes
- B09B3/65—Anaerobic treatment
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
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Abstract
The invention discloses a method for strengthening anaerobic treatment of kitchen waste by using a conductive carrier to form a Shewanella electroactive biomembrane. The method for forming the electroactive biological film comprises the following steps: (1) the conductive carbon cloth is soaked in ethanol and acetone with the proportion of 1:1, carrying out ultrasonic treatment for 2 hours, taking out, washing with clear water and air-drying; (2) Soaking the conductive carbon cloth treated in the step (1) in 0.5-1.5M nitric acid solution and sulfuric acid solution for 2h respectively, taking out and drying; (3) Spreading the conductive carbon cloth treated in the step (2), soaking in Shewanella bacterium solution containing 0.5-1.5M lactose, standing for 1d, taking out, air-drying at 30+/-2 ℃ and pH of the bacterium solution within the range of 6.5+/-0.5, and repeating the steps for 2-3 times to form the electroactive biological film. The invention utilizes the Shewanella electroactivity biological film formed on the surface of the conductive carrier to treat kitchen waste, accelerates the electron transfer rate in the anaerobic reaction process, improves the anaerobic digestion efficiency, solves the acid accumulation problem in the anaerobic digestion process, and reduces the chemical treatment and operation cost.
Description
Technical Field
The invention particularly relates to a method for strengthening anaerobic treatment of kitchen waste by using a conductive carrier to form a Shewanella electroactive biomembrane, belonging to the technical field of organic waste treatment and recycling.
Background
Kitchen garbage which is used as a main component of the household garbage contains a large amount of organic matters, and has high recycling value. The daily kitchen waste disposal problem is increasingly outstanding, and simultaneously, a huge market is brought, and the aspects of recycling and harmless treatment of kitchen waste become the focus of attention of all parties.
At present, the main means for treating kitchen waste in China is a wet anaerobic digestion technology. Compared with physical and chemical methods, the anaerobic digestion technology has the advantages of low device cost, no secondary pollution, effective resource recycling and the like. In the anaerobic kitchen waste digestion process, organic components in kitchen waste are decomposed into substances such as micromolecular organic acid, alcohol and the like through the hydrolytic fermentation of microorganisms, and then the micromolecular acid and the alcohol are decomposed into acetic acid, hydrogen and CO through the hydrogen production and acetic acid production process 2 Finally converting acetic acid into methane and CO under the action of acetic acid nutritional methane bacteria 2 Hydrogen and CO under the action of hydrogenotrophic methane bacteria 2 Converted to methane and water. However, kitchen waste has complex components and is easy to acidify, the hydrolysis fermentation process is fast in speed, and the volatile acid generated by degrading the kitchen waste cannot be digested and degraded in time in the methane production process during high-load operation, so that the phenomenon of accumulation of the volatile acid occurs, the activity of methanogen is inhibited, and the efficient treatment of the kitchen waste is influenced. Adding strong alkali to adjust pH is a common means for solving the problem of acid accumulation, and can provide a proper growing environment for methane production in an anaerobic system, but increase kitchen waste treatment cost, and meanwhile, adding strong alkali for a long time can lead to system salinity improvement, can also influence the growth of microorganisms, and is unfavorable for long-term stable operation of the system.
Disclosure of Invention
The invention provides a method for coupling Shewanella with a conductive carrier and application of the method in kitchen waste treatment.
The invention has the innovation points that the electric carrier of the Shewanella electric activity biological film is formed on the surface of the electric carrier by adding the electric carrier on the surface of the anaerobic device, the Shewanella electric activity biological film is formed on the surface of the electric carrier, so that the Shewanella electric activity biological film is convenient to fix and utilize the Shewanella and the flavins secreted by the Shewanella electric activity biological film, meanwhile, the electric activity fermentation bacteria and the electric activity methanogen capable of directly receiving electron methanogenesis are easier to enrich on the Shewanella electric activity biological film, and the electric activity biological film is used as a medium for electron transfer between microorganisms to improve the electron transfer rate of anaerobic digestion and strengthen the anaerobic digestion effect of kitchen waste.
According to the invention, organic matter is fermented and acidogenic to be decomposed into micromolecular substances by using Shewanella, and the secreted flavine can specifically strengthen the methanogenic process of the electroactive methanogenic bacteria, the conductive carrier is used for fixing the flavine and Shewanella to further promote the rate of transferring electrons by surface microorganisms and ensure a certain amount of flavine with certain concentration, so that the dynamic balance of the hydrolysis fermentation and methanogenic process in the anaerobic digestion treatment of high-load kitchen waste is maintained, and meanwhile, the loss or continuous addition of the flavine is avoided.
In order to achieve the above object, the present invention provides a method for forming an electroactive biofilm of Shewanella on the surface of a conductive carrier.
The method for forming the Shewanella electroactivity biological film on the surface of the conductive carrier provided by the invention comprises the following steps:
1) Preparing Shewanella bacteria liquid;
2) Soaking the pretreated conductive carrier in Shewanella bacteria liquid obtained in the step 1) after adding 0.5-1.5M lactose, thereby forming the Shewanella electroactive biological film on the surface of the conductive carrier.
The operation of the method step 1) is as follows: inoculating Shewanella bacteria strain into a liquid culture medium for culture to obtain Shewanella bacteria liquid;
wherein the Shewanella has a flavine secretion capability, and can be a commercially available Shewanella Shewanella oneidensis strain; the Shewanella bacteria agent purchased by the commercial platform is Shewanella oneidensis MR-1 bacteria agent Shewanella bacteria concentration is 1 multiplied by 10 8 ～4×10 8 Individual/ml
The liquid culture medium is prepared from peptone, naCl, yeast extract powder, agar powder and deionized water; specifically, the liquid culture medium is prepared from the following components: 5g peptone, 5g NaCl, 2.5g yeast extract, 500ml deionized water.
Each 100. Mu.g of Shewanella (number of viable bacteria 1X 10) 5 ～4×10 5 Mu.g) to 100ml of the liquid medium;
the temperature of the culture can be 28-32 ℃, specifically 30 ℃, and the time can be 0.8-1.5 days, specifically 1 day;
the OD600 of the obtained Shewanella liquid is kept above 0.36;
in the step 2 of the method, the conductive carrier may be any one of conductive carbon cloth, carbon felt and carbon brush;
specifically, the conductive carbon cloth can be 360 μm thick and 125g/m 2 Conductive carbon cloth with density and resistivity less than 2Ω·m;
the pretreatment comprises cutting, cleaning and acid soaking treatment;
the cleaning operation is as follows: soaking the cut conductive carrier in a mixed solution of ethanol and acetone, performing ultrasonic treatment, taking out and air-drying;
wherein, the volume ratio of ethanol to acetone in the mixed solution can be 1:1, a step of;
the pretreatment can further comprise the operation of washing the air-dried conductive carrier with clear water for 2-3 times, soaking in the clear water and washing out scum;
the acid soaking operation is as follows: soaking the conductive carrier in 0.5-1.5M nitric acid solution for 1-3h; soaking in 0.5-1.5M sulfuric acid solution for 1-3 hr, taking out and stoving;
the temperature of the acid soaking can be specifically 30+/-2 ℃.
In the step 2), the pretreated conductive carrier is soaked in Shewanella bacteria liquid for a plurality of times, specifically for 2-3 times, the soaking time is 1d, and the air drying operation is carried out after each soaking;
the soaking temperature is 30+/-2 ℃.
The pH of the Shewanella bacteria liquid is within the range of 6.5+/-0.5.
The application of the conductive carrier with the Shewanella electro-active biological film formed on the surface in anaerobic treatment of kitchen waste also belongs to the protection scope of the invention.
The invention also provides a method for strengthening anaerobic treatment of kitchen waste by using the Shewanella electro-active biomembrane formed by the conductive carrier.
The invention provides a method for strengthening anaerobic treatment of kitchen waste by using a conductive carrier to form a Shewanella electroactive biomembrane, which comprises the following steps:
the conductive carrier with the Shewanella electro-active biological film formed on the surface, which is prepared by the method, is put into a kitchen waste anaerobic treatment device for anaerobic digestion treatment of kitchen waste;
wherein Shewanella bacteria liquid (Shewanella bacteria liquid obtained in the step 1 after adding 0.5M to 1.5M lactose) is periodically added in the initial period of operation.
In the kitchen waste anaerobic treatment method, anaerobic granular sludge and cow dung in a beer waste water treatment plant are uniformly mixed to be used as inoculation sludge, and the mass ratio of the sludge to the cow dung is 3:1-5:1;
the anaerobic granular sludge is a term well known in the art, and specifically includes anaerobic sludge for treating high-concentration COD wastewater.
The conductive carrier is added into the anaerobic treatment device, and the mass ratio of the conductive carrier to the anaerobic granular sludge is 4-5: 100;
the Shewanella bacteria liquid is added with 1/1000 of the volume of the anaerobic treatment device every 3d in the initial operation period,
the adding is repeated for a plurality of times, and the specific time can be 2-3 times.
The kitchen waste is fed into the anaerobic treatment device at the initial stage of operation, and the mass ratio of the kitchen waste to the anaerobic granular sludge is 1: and 50, after gas production occurs, the organic load of the device is lifted periodically.
The organic load is lifted after the device reaches the removal rate of more than 80 percent, and the lifting degree is not more than 50 percent of the previous load.
The kitchen waste is crushed until the granularity is less than 5mm before anaerobic digestion, and is uniformly mixed.
The anaerobic digestion is carried out at a ph=7.0±1.0 and a temperature of 30 ℃ to 35 ℃.
The invention combines Shewanella with conductive carbon cloth to play a coupling role in the kitchen waste anaerobic device. Mixing the Shewanella agent with conductive carbon cloth and kitchen waste, and performing anaerobic digestion treatment to realize efficient treatment of the kitchen waste.
The invention has the following advantages:
(1) The conductive carbon cloth is pretreated by acid soaking, the specific surface area of the carbon cloth is increased, the conductivity is increased, and hydrophilic functional groups are attached. In kitchen waste treatment application, a microorganism growth environment and an electron transfer path can be provided, and an anaerobic digestion system taking electron transfer between direct inoculation as a main metabolic pathway is formed.
(2) The Shewanella provided by the invention can generate a certain amount of flavine accumulation within 72-96 hours when entering an anaerobic device, the flavine specificity strengthens electron transfer functional proteins, the methanogenic path of electron transfer (Direct interspecies electron transfer, DIET) between direct inoculation of methanogens is improved, and the device can maintain the contribution of the DIET for a long time under the condition of not adding the flavine.
(3) The invention combines the conductive carbon cloth and the Shewanella agent (as shown in figure 1), can utilize the electroactive biomembrane of the electrogenerated bacteria Shewanella of various organic matters to be more easily attached on the surface of the conductive carbon cloth, reduces the distance of the electron transfer process by the capillary action of the biomembrane, and accelerates the degradation of the organic matters and the electron transfer.
(4) The coupling technology of the invention has simple operation, convenient management and good application prospect and economic benefit in kitchen waste treatment application.
Drawings
FIG. 1 is a schematic diagram of a conductive carbon cloth for forming an electroactive biofilm of Shewanella in accordance with the present invention. The individual labels are as follows: 1 is conductive carbon cloth; 2 is an electroactive biological film; 3 is Shewanella; 4 is flavin.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The Shewanella preparation used in the following examples was Shewanella oneidensis MR-1 bacteria and Shewanella concentration was 1X 10 8 ～4×10 8 Individual/ml
EXAMPLE 1 preparation of Shewanella liquid
Preparation of LB liquid Medium: 5g peptone, 5g NaCl, 2.5g yeast extract, 500ml deionized water in a 1000ml beaker. Placing into a rotor, stirring with a magnetic stirrer, dissolving, mixing thoroughly (2 h), packaging into 5 dried 250ml conical flasks, and sealing with tinfoil paper. Sterilizing at 120deg.C for 30min, and storing at room temperature for 2 weeks. 100 μg (number of viable bacteria 1×10) 5 ～4×10 5 Mu.g) Shewanella and 100ml of liquid medium, and culturing at 30 ℃ for 1 day after mixing, so that the OD600 of the liquid is kept above 0.36.
EXAMPLE 2 formation of Shewanella electroactive biofilm on the surface of an electrically conductive Carrier
(1) Conductive carbon cloth (360 μm thickness, 125 g/m) 2 Density, resistivity < 2Ω·m) into cloth with length, width and height of 20cm×20cm, soaking in ethanol and acetone with volume ratio of 1:1 and ultrasonically treating for 2 hours, taking out, washing 3 times with clear water and air-drying. Soaking the carbon cloth in a 1M nitric acid solution for 2 hours and in a 1M sulfuric acid solution for 2 hours, taking out and drying the carbon cloth, wherein the temperature can be 30+/-2 ℃; the conductive carbon cloth is a 3K carbon fiber cloth lining interlayer cloth purchased from Carbonium store of IVA family of Taobao platform.
(2) Pouring Shewanella bacteria liquid prepared in example 1 containing 0.5M lactose into a cuboid container, spreading conductive carbon cloth in the container in sequence, ensuring that the liquid level is 1cm above the surface of the carbon cloth, standing for 1d, taking out, air drying at 30+/-2 ℃ and the pH of the bacteria liquid being in the range of 6.5+/-0.5. The carbon cloth is soaked in Shewanella bacteria liquid for 1d again, a biological film is further formed, and the carbon cloth is put into an anaerobic device after being air-dried.
Example 3 anaerobic treatment of kitchen waste
The experiment is carried out by adopting a wet anaerobic digestion process, and the specific steps are as follows:
(1) The inoculated sludge (mixture of anaerobic granular sludge and cow dung, mass ratio of sludge to cow dung is 3:1) with solid content (mass percent of solid) of 10% and the conductive carbon cloth obtained by processing in example 2 are added into anaerobic fermentation equipment, and anaerobic fermentation is carried out under the environment of initial pH of 7.0 and temperature of 35 ℃. Shewanella bacteria liquid prepared in example 1 containing 0.5M lactose was added to the anaerobic apparatus every 3d at the initial stage of operation, and the volume ratio of the Shewanella bacteria liquid to the apparatus was 1:1000 repeating the step 3 times;
(2) Operation of kitchen waste treatment device: at 1.5kg COD/(m) 3 D) starting load, and gradually increasing the load according to fermentation parameters such as COD, ammonia nitrogen, pH value and the like in the fermentation liquid in the later period; the organic load is adjusted by adjusting the feeding amount, and the device cannot absorb 50% of the feeding amount, namely the experiment is ended. And (3) experimental operation 120d, and observing various water quality indexes under a stable operation state.
Comparative example 1
Only in example 3, the Shewanella agent was not added to the apparatus during the start-up period (there was no "Shewanella liquid prepared in example 1 was added to the anaerobic apparatus every 3d in the initial stage of operation"), but the same procedure was adopted.
Comparative example 2
Only in example 3, the conductive carbon cloth (the conductive carbon cloth is not pretreated) and the inoculated sludge are added to the device at the same time in the starting period, shewanella bacteria liquid is not added, and the same operation is adopted for the other parts.
Comparative example 3
Only in example 3, the Shewanella agent was added to the apparatus at the same time during the start-up period (Shewanella liquid prepared in example 1 was added to the anaerobic apparatus every 3d at the initial stage of operation, the volume ratio to the apparatus was 1:1000, and this step was repeated 3 times) and the inoculated sludge, and the same operation was adopted for the other.
Comparative example 4
Only in example 3, the inoculation sludge was fed to the apparatus only during the start-up period, the same procedure being followed.
The detection of the change of indexes such as COD, pH, volatile acid, methane and the like in the anaerobic kitchen waste digestion process of the example 3 and the comparative example shows that the organic matter removal load of the example 3 is up to 9.0 gCOD/(m) 3 D) 13% higher than comparative example 1, 29% higher than comparative example 2, 39% higher than comparative example 3, and 64% higher than comparative example 4. The pH of example 3 was maintained between 6.8 and 7.5, while the pH of comparative example 4 was gradually reduced to 5.5 at high organic load. Meanwhile, the volatile acid content of example 3 was 65%, 42%, 33%, 20% of the comparative example content, respectively. The methane conversion of example 3 reaches 520ml CH 4 and/gVS, 114% higher than the comparative example with the highest conversion.
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains.
Claims (4)
1. A method of forming a shiwanella electroactive biofilm on a surface of a conductive support, comprising the steps of:
1) Preparing Shewanella bacteria liquid;
2) Soaking the pretreated conductive carrier in Shewanella bacteria liquid obtained in the step 1) after adding 0.5-1.5M lactose, so as to form an electric active biological film of Shewanella bacteria on the surface of the conductive carrier;
the conductive carrier is conductive carbon cloth;
the pretreatment comprises cutting, cleaning and acid soaking treatment;
the cleaning operation is as follows: soaking the cut conductive carrier in a mixed solution of ethanol and acetone, performing ultrasonic treatment, taking out and air-drying;
wherein the volume ratio of ethanol to acetone in the mixed solution is 1:1, a step of;
the acid soaking operation is as follows: firstly, soaking a conductive carrier in 0.5-1.5M nitric acid solution for 1-3h; soaking in 0.5-1.5M sulfuric acid solution for 1-3h, taking out and drying;
the temperature of the acid soaking is 30+/-2 ℃;
in the step 1), the Shewanella strain is inoculated into a liquid culture medium for culture, so as to obtain Shewanella bacterial liquid;
the Shewanella strain isShewanella oneidensisMR-1 bacteria;
the liquid culture medium is prepared from peptone, naCl, yeast extract powder and deionized water;
every 100 mu g of Shewanella, the number of viable bacteria is 1 multiplied by 10 5 ~4×10 5 Inoculating to 100ml of the liquid culture medium;
the culture temperature is 28-32 ℃ and the culture time is 0.8-1.5 days;
the liquid OD600 of the Shewanella liquid was maintained at 0.36 or more.
2. The method according to claim 1, characterized in that: in the step 2), the pretreated conductive carrier is soaked in Shewanella bacteria liquid for a plurality of times, specifically for 2-3 times, the soaking time is 1d, and the air drying operation is carried out after each soaking;
the soaking temperature is 30+/-2 ℃;
the pH of the Shewanella bacteria liquid is within the range of 6.5+/-0.5.
3. Use of a conductive carrier with a Shewanella electroactive biofilm formed on the surface thereof, prepared by the method of claim 1 or 2, in anaerobic treatment of kitchen waste.
4. A kitchen waste anaerobic treatment method comprises the following steps: feeding the conductive carrier with the Shewanella electroactive biomembrane formed on the surface, which is prepared by the method of claim 1 or 2, into a kitchen waste anaerobic treatment device for anaerobic digestion treatment of kitchen waste,
wherein Shewanella bacteria liquid is periodically added in the initial period of operation;
in the method, anaerobic granular sludge and cow dung in a beer wastewater treatment plant are uniformly mixed to be used as inoculation sludge, and the mass ratio of the sludge to the cow dung is 3:1-5:1;
the conductive carrier is added into the anaerobic treatment device, and the mass ratio of the conductive carrier to the anaerobic granular sludge is 4-5: 100;
the anaerobic digestion treatment is carried out at the pH=7.0+/-1.0 and the temperature of 30-35 ℃; the Shewanella bacteria liquid is added with 1/1000 of the volume of the anaerobic treatment device every 3d in the initial operation period;
the adding is repeated for a plurality of times, specifically 2-3 times.
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| CN112047590A (en) * | 2020-08-31 | 2020-12-08 | 同济大学 | Method for strengthening anaerobic digestion of sludge by utilizing pre-alcoholization of kitchen waste |
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