CN102876724A - Coupling method based on synchronous cellulose enzymolysis and fermentation and microbial electrolysis cell hydrogen production - Google Patents
Coupling method based on synchronous cellulose enzymolysis and fermentation and microbial electrolysis cell hydrogen production Download PDFInfo
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- CN102876724A CN102876724A CN2012103699979A CN201210369997A CN102876724A CN 102876724 A CN102876724 A CN 102876724A CN 2012103699979 A CN2012103699979 A CN 2012103699979A CN 201210369997 A CN201210369997 A CN 201210369997A CN 102876724 A CN102876724 A CN 102876724A
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Abstract
The invention discloses a coupling method based on synchronous cellulose enzymolysis and fermentation and microbial electrolysis cell hydrogen production. The coupling method is characterized by including the steps: A, uniformly mixing unpretreated or pretreated cellulose matrix and combined enzyme liquid according to the ratio of 100: (0.5-1) and adding the mixture into a sterilized electrolysis cell; B, adding anaerobic fermentation bacteria or photosynthetic bacteria or mixed bacteria of the anaerobic fermentation bacteria and the photosynthetic bacteria into the electrolysis cell; C, adding defined medium liquid into the electrolysis cell; and D, externally connecting the electrolysis cell with a resistor, starting the electrolysis cell, switching in an external power supply, adjusting input voltage to range from 0.2 volt to 1.2 volts and keeping the electrolysis cell in a hydrogen production mode. The addition of the mixture of the cellulose matrix and the combined enzyme liquid is 10%-20% of the volume of the electrolysis cell, the inoculation amount of the bacteria is 5%-35% (v/v), and the addition of the defined medium liquid is 10%-35% of that of the added matrix.
Description
Technical field
The present invention relates to microorganism electrolysis cell and produce hydrogen methods, be specifically related to utilize the synchronous enzymatic hydrolysis and fermentation of cellulose substances and microorganism electrolysis cell to produce the coupling process of hydrogen.
Background technology
Current, along with the aggravation of countries in the world energy dilemma, the biomass material take cellulose as major ingredient has obtained studying widely and utilizing in various countries as renewable energy source.Have data to show, the total amount of the annual production of cellulose in the whole world and hemicellulose is about 85,000,000,000 tons, and Mierocrystalline cellulose extensively is present in the plants such as straw, stalk, tree bar as compound of polysaccharide.There is a large amount of agricultural wastes every year in China as large agricultural country, and it can be used as the renewable resources of cheapness.From 2005, the straw ultimate production in China every year reached 8.42 hundred million tons, and wherein straw reaches 1.8 hundred million tons, if with its whole utilizations, will alleviate the energy dilemma of China.
In the cellulose hydrolysis saccharifying, hydrolysate has the materials such as dextran, xylan, acetic acid.If only utilize saccharification of cellulose glucose later, this just causes the waste of resource, and can cause environmental pollution.Based on the recycling rate that improves organic waste and the purpose of products collection efficiency, need the cellulose hydrolysis saccharified liquid is fully utilized, and minimizing is to the pollution of environment.
At present, adopting biomass is the method that raw material hydrogen manufacturing mainly adopts enzymolysis and fermentation to carry out respectively, and the problem that mainly exists for this method has: the hydrogen and the CO that 1. produce
2Mix Deng gas, so the purity of hydrogen is not high in the gas; 2. because the biomass enzymolysis process is to separate to carry out with fermenting process, so need to adopt two covering devices, this has just caused the increase of initial stage input cost; 3. biomass enzymatic hydrolysis and fermentation proceed step by step, the hydrogen manufacturing cycle stretch-out; 4. enzymolysis and fermentation and hydrogen production cause respectively the reducing sugar accumulation of cellulose hydrolysis generation and saccharifying are produced stronger restraining effect.
Adopt the technology of synchronous enzymatic hydrolysis and fermentation hydrogen manufacturing to overcome the deficiency of above-mentioned fractional hydrolysis hydrogen manufacturing, the technology of synchronously enzymatic hydrolysis and fermentation hydrogen manufacturing has the advantage of following several respects: 1. synchronously enzymatic hydrolysis and fermentation hydrogen manufacturing has realized carrying out synchronously of fermenting process and hydrogen production process, can shorten the hydrogen manufacturing cycle; 2. synchronously enzymatic hydrolysis and fermentation hydrogen manufacturing is carried out in same reactor, and hydrogen manufacturing is simple to operate, has reduced production cost; 3. synchronously enzymatic hydrolysis and fermentation hydrogen manufacturing can be eliminated the product inhibition in enzymolysis process and the hydrogen production process; 4. synchronous enzymatic hydrolysis and fermentation hydrogen producing technology is applied in the microorganism electrolysis cell, all kinds of materials that cellulose hydrolysis produces comprise that some inhibition products can be utilized generation hydrogen by microorganism, can obviously improve and produce the hydrogen yield.
Summary of the invention
Technical problem to be solved by this invention is the coupling that the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell produce hydrogen.
In order to solve the problems of the technologies described above, technical scheme of the present invention is that the coupling process based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen is characterized in that:
A. incite somebody to action not pre-treatment or pretreated cellulose matrix and make up enzyme liquid according to 100:(0.5～1) mix, and with in the electrolyzer after the mixed solution adding sterilising treatment of cellulose matrix and combination enzyme liquid, add-on is 10%～20% of electrolyzer volume;
B. anaerobically fermenting bacterium, photosynthetic bacterium or the mixed bacterium of the two are added electrolyzer, inoculum size is 5%～35% (v/v).The inoculated bacteria purpose is to utilize these microorganisms to decompose the organism such as reducing sugar that the mixed enzyme solution hydrolysis produces, and produces hydrogen;
C. in electrolyzer, add the combination medium liquid, add-on by adding matrix 10%～35%;
D. the electrolyzer outer meeting resistance starts electrolyzer, after electrolyzer starts successfully, and the access external power source, regulating input voltage is 0.2～1.2 volt, electrolyzer is in the hydrogen pattern of producing; At H
2The collection and confinement of gases outlet is collected H by gas collector
2Gas is at CO
2The collection and confinement of gases outlet is collected CO by gas collector
2Gas, treat that substrate consumption is complete after, waste liquid is discharged by leakage fluid dram 4.
Principle of the present invention is: Mierocrystalline cellulose and hemicellulose resolve into dextran and xylan under the mixed enzyme solution effect of cellulase, beta-glucosidase, zytase, cellulose matrix also produces by product in the enzymatic saccharification process, such as acetic acid, furfural, carboxymethyl furfural etc.The product of the enzymatic saccharification processes such as glucose, acetic acid generates CO under the effect of inoculated bacteria simultaneously
2, H
+And electronics, H
+To cathode motion, electronics conducts to negative electrode along external circuit under the effect of potential difference, H
+With come to be combined at negative electrode to the electronics of external circuit reduction reaction occurs to generate H
2, then collect H at the cathode side top
2, collect CO at the anode side top
2Thereby,, realized simultaneously that in microorganism electrolysis cell cellulase hydrolysis effect and microbial fermentation produce hydrogen.
Utilize the method for the synchronous enzymatic hydrolysis and fermentation hydrogen manufacturing of microorganism electrolysis cell, so that under the effect of certain impressed voltage, anaerobically fermenting bacterium or photosynthetic bacterium can in time utilize the material of reducing sugar one class of enzymolysis generation to do substrate and produce the hydrogen metabolism, but also can utilize the inhibitory substance in the hydrolyzed solution to produce hydrogen, improved significantly hydrogen generation efficiency, thereby reduced the pollution to environment, realized the efficient utilization of resource.
The system response equation:
Cellulose hydrolysis changes into the reaction formula of glucose:
(C
6H
10O
5)
n+nH
2O?→nC
6H
12O
6
The reaction formula of glucose in microorganism electrolysis cell:
Anode: C
6H
12O
6+ 6H
2O → 6CO
2+ 24H
++ 24e
-
Negative electrode: 24H
++ 24e
-→ 12H
2
The reaction formula of acetic acid in microorganism electrolysis cell:
Anode: C
2H
4O
2+ 2H
2O
2→ 2CO
2+ 8H
++ 8e
-
Negative electrode: 8H
++ 8e
-→ 4H
2
Produce the preferred version of the coupling process of hydrogen according to of the present invention based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell, when microorganism electrolysis cell adopted the single-chamber microbial electrolyzer, steps A, material that B, C add were in the direct adding electrolyzer.
According to the preferred version that produces the coupling process of hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell of the present invention, when microorganism electrolysis cell adopts the double-chamber microbiological electrolyzer, in steps A, the anolyte compartment of material that B, C add for the adding electrolyzer, and before step D, also carry out following steps:
Be 20～60mML with concentration
-1Potassium ferricyanide solution or concentration be 20～60mML
-lPotassium sulfite solution add in the cathode compartment of electrolyzer, and equate with the volume of the added solution in anolyte compartment at the volume of solution that cathode compartment adds.
According to the preferred version that produces the coupling process of hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell of the present invention, when microorganism electrolysis cell adopts the double-chamber microbiological electrolyzer, the mixed bacterium or single photosynthetic bacterium or the anaerobically fermenting bacterium that also add simultaneously anaerobically fermenting bacterium and photosynthetic bacterium in the cathode of electrolytic tank chamber, inoculum size is 5%～35% (v/v), and equates with the volume of the added solution in anolyte compartment at the volume of solution that cathode compartment adds.
Produce the preferred version of the coupling process of hydrogen according to of the present invention based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell, when microorganism electrolysis cell adopts the double-chamber microbiological electrolyzer, in cathode compartment, also add pH and be 4.8, concentration is 40～80mML
-1The phosphoric acid salt sustained release dosage, the ratio of the phosphoric acid salt sustained release dosage that adds and potassium ferricyanide solution or potassium sulfite solution is 4: 1; And the volume at solution that cathode compartment adds equates with the volume of the added solution in anolyte compartment.
According to the preferred version that produces the coupling process of hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell of the present invention, described enzyme liquid is cellulase, beta-glucosidase, zytase, mixes in 22～40mg: 0.02～0.08mL:10～28mg ratio.Wherein, cellulase energy hydrocellulose β-1,4 glycosidic links, make cellulose decomposition become cellobiose and glucose, beta-glucosidase can be decomposed into glucose with cellobiose and other low molecule cellodextrins, zytase can resolve into the small-molecule substance wood sugar with contained xylan in the biomass class material, the mixed enzyme solution of cellulase, beta-glucosidase, zytase can become small-molecule substance with material decomposition such as the Mierocrystalline cellulose in the biomass, cellobiose, xylans, finally is hydrolyzed into glucose.Therefore adding the performance that mixed enzyme solution adds the single enzyme preparation gets a promotion, various effective constituents in the hydrocellulose matrix more effectively, significantly improve the transformation efficiency that Wood Adhesives from Biomass becomes glucose, realization is to the comprehensive utilization of various compositions in the biomass, thereby obtains more reducing sugar.
Produce the preferred version of the coupling process of hydrogen according to of the present invention based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell, described combination substratum is: 5.8～6.2gL
-1Na
2HPO
4, 2.8～3.2gL
-1KH
2PO
4, 0.08～0.12gL
-1CO (NH
2)
2, 0.5～0.7gL
-1NaCl, 0.2～0.24gL
-1MgSO
47H
2O, 14～16mgL
-1CaCl
22H
2O and 6～12mgL
-1Trace element solution, wherein trace element is: Mo, Fe, Zn, Co or Cu.
The beneficial effect that produces the coupling process of hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell of the present invention is: the present invention carries out biological hydrogen production with the synchronous enzymatic hydrolysis and fermentation of cellulose matrix and microorganism electrolysis cell combination, have the following advantages: 1. matrix enzymatic saccharification efficient improves, metabolic intermediate can be fully used, and energy consumption is low; 2. can realize producing hydrogen and produce CO
2Separation has spatially greatly improved hydrogen purity, has reduced the expense of later stage separating-purifying; 3. carrying out synchronously of enzymatic saccharification process, anaerobically fermenting and biological hydrogen production effect effectively shortened the production cycle; 4. carry out synchronously enzymatic hydrolysis and fermentation and biological hydrogen production in the electrolyzer reactor, equipment is simple, and is easy to operate, reduced the input of first current cost.The present invention has realized the efficient purpose that produces hydrogen when processing farming, woods waste, has the advantage of green, energy-saving and environmental protection, has a good application prospect.
Description of drawings
Fig. 1 is that the coupling process based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen of the present invention adopts single-chamber microbial electrolyser construction synoptic diagram.
Fig. 2 is that the coupling process based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen of the present invention adopts double-chamber microbiological electrolyser construction synoptic diagram.
Embodiment
Embodiment 1:
Coupling process based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen comprises the steps:
The first step, making single-chamber microbial electrolyzer: comprise anode, negative electrode.Above cathode side, be provided with H
2The collection and confinement of gases outlet is provided with CO above anode utmost point side
2The collection and confinement of gases outlet is provided with opening for feed 3 in anode side, is provided with leakage fluid dram 4 at cathode side;
Second step, electrolyzer sterilising treatment: at first with 30% formalin solution electrolyzer is sterilized, and with cleaning through the deionized water that cools off behind the high-temperature sterilization, guarantee that the mixed bacterium of anaerobically fermenting bacterium or photosynthetic bacterium or anaerobically fermenting bacterium and photosynthetic hydrogen production bacterium is not by other bacterial contamination, and be in good living environment, then applying argon gas guarantees that electrolyzer keeps the closing anaerobic state;
The 3rd step, cellulose matrix pre-treatment: rice straw is cut into 1～2cm segment, clean and dry to constant weight, getting the rice straw of cleaning oven dry utilizes pulverizer to pulverize, utilize automatic vibrating screen to cross 40～120 order numbers, take by weighing the aseptic straw stalk powder of 20～80g and put into the beaker of 1000mL, then add 0.5～2.0%NaOH solution 500mL in the lower 24h of placement of room temperature (30 ℃).After finishing dealing with, with distilled water rice straw is washed, until distilled water pH value is neutral after the flushing, flushing to neutral rice straw matrix is dried to constant weight in 105 ℃ of lower baking ovens, get the rice straw powder in high-pressure sterilizing pot in 121 ℃ of lower sterilization 10min, then take out stand-by.The matrix of rich cellulose class material can also adopt: the biomass such as maize straw, wheat straw waste, tree bar;
In addition, cellulosic pretreatment process also comprises sour pre-treatment, ultrasonic pretreatment, high temperature steaming pre-treatment etc.
The 4th goes on foot, pretreated cellulose matrix and enzyme liquid is mixed according to 100:0.5, and carry out sterilising treatment, the mixed solution of the cellulose matrix after the sterilization and enzyme liquid is added electrolyzer by opening for feed 3, the volume of the mixed solution of the cellulose matrix that adds and enzyme liquid accounts for 20% of electrolyzer volume, described enzyme liquid is cellulase, beta-glucosidase, zytase, and in 22mg: the 0.02mL:10mg ratio is mixed;
The 5th step, with the mixed bacterium of anaerobically fermenting bacterium and photosynthetic bacterium by opening for feed 3 adding electrolyzers, inoculum size is 20% (v/v);
The 6th step, add the combination medium liquids by opening for feed 3, add-on by adding matrix 10%, the prescription that wherein makes up substratum is: 6.2gL
-1Na
2HPO
4, 2.8gL
-1KH
2PO
4, 0.1gL
-1CO (NH
2)
2, 0.5gL
-1NaCl, 0.2gL
-1MgSO
27H
2O, 15mgL
-1CaCl
22H
2O and 10mgL
-1FeSO
47H
2O solution;
The 7th step, the external 100 Ω pull-up resistors of electrolyzer start electrolyzer.The electrolyzer initial start stage is 0～4d, and the load both end voltage is about 0mV, and passing in time, and bacterium forms microbial film at anode surface, and voltage slowly increases.After starting 5～8d, voltage quickenings that gather way is Exponential growth, until biofilm development is stable, inoculate about 7～10d after, load voltage tends towards stability gradually, this shows that start battery successfully;
After the 8th step, electrolyzer start successfully, the access power supply, regulating input voltage is 0.2～1.2 volt, electrolyzer is in the hydrogen pattern of producing.More H
2Discharge the H of generation at negative electrode
2Collect by cathode side venting port collection device, at the more CO of anode side
2Gas is produced, and collects CO by the anode side gas collection device
2Gas, treat that substrate consumption is complete after, waste liquid is discharged by leakage fluid dram 4;
Conclusion: by the coupling of the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen, so that collected H
2Purity is high, can reach more than 70%, has reduced the process of later stage gas delivery, purifying, has reduced production cost.
Embodiment 2:
Coupling process based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen comprises the steps:
The first step, making double-chamber microbiological electrolyzer: comprise anolyte compartment, cathode compartment, the centre separates with ion-exchange membrane; Be provided with H at the cathode compartment top
2The collection and confinement of gases outlet is provided with CO at the top, anolyte compartment
2The collection and confinement of gases outlet is provided with leakage fluid dram 6 and leakage fluid dram 4 in the bottom of cathode compartment and anolyte compartment, be provided with fluid inlet 5 and opening for feed 3 on the top of cathode compartment and anolyte compartment;
Second step, electrolyzer sterilising treatment: at first with 35% formalin solution electrolyzer is sterilized, and with cleaning through the deionized water that cools off behind the high-temperature sterilization, the single photosynthetic hydrogen production bacterium that assurance cathode compartment anolyte compartment adds or the mixed bacterium of anaerobically fermenting bacterium or anaerobically fermenting bacterium and photosynthetic bacterium be not by other bacterial contamination, and be in good living environment, then applying argon gas guarantees that electrolyzer keeps the closing anaerobic state;
The 3rd step, cellulose matrix pre-treatment: rice straw is cut into 1～2cm segment, clean and dry to constant weight, getting the rice straw of cleaning oven dry utilizes pulverizer to pulverize, utilize automatic vibrating screen to cross 40～120 order numbers, take by weighing the aseptic straw stalk powder of 20～80g and put into the beaker of 1000mL, then add 0.5～2.0%NaOH solution 500mL in the lower 24h of placement of room temperature (30 ℃).After finishing dealing with, with distilled water rice straw is washed, until distilled water pH value is neutral after the flushing, flushing to neutral rice straw matrix is dried to constant weight in 105 ℃ of lower baking ovens, get the rice straw powder in high-pressure sterilizing pot in 120 ℃ of lower sterilization 10min, then take out stand-by.Cellulose matrix can also adopt: the biomass such as maize straw, wheat straw waste, tree bar.
In addition, the pretreatment process of cellulose matrix also comprises sour pre-treatment, ultrasonic pretreatment, high temperature steaming pre-treatment etc.;
The 4th goes on foot, pretreated cellulose matrix and enzyme liquid is mixed according to 100:1, and carry out sterilising treatment, the anolyte compartment that the mixed solution of the cellulose matrix after the sterilization and enzyme liquid is added electrolyzer by the opening for feed 3 of anolyte compartment, the mixed solution of the cellulose matrix that adds and enzyme liquid accounts for 10% of electrolytic cell anode chamber volume, described enzyme liquid is cellulase, beta-glucosidase, zytase, and in 40mg: the 0.08mL:28mg ratio is mixed;
The 5th step, the opening for feed 3 of photosynthetic bacterium by the anolyte compartment added the anolyte compartment of electrolyzers, inoculum size is 20%v/v;
The 6th step, the opening for feed by the anolyte compartment 3 add the combination medium liquids, add-on by adding matrix 30%, the prescription that wherein makes up substratum is: 6.0gL
-1Na
2HPO
4, 3.0gL
-1KH
2PO
4, 0.08gL
-1CO (NH
2)
2, 0.6gL
-1NaCl, 0.22gL
-1MgSO
47H
2O, 14mgL
-1CaCl
22H
2O and 10mgL
-1(NH
4)
6Mo
7O
244H
2The solution of O;
The 7th step, be 50mML with concentration
-1The fluid inlet 5 of potassium ferricyanide solution by cathode compartment add cathode compartments, and to add simultaneously pH be that 4.8 concentration are 60mML
-1The phosphoric acid salt sustained release dosage, the phosphoric acid salt sustained release dosage that adds and the ratio of potassium ferricyanide solution are 4:1; The volume of the solution that adds at cathode compartment equates with the volume of the added solution in anolyte compartment;
The 8th step, electrolyzer external load resistance start electrolyzer.The electrolyzer initial start stage is 2d, and the load both end voltage is about 0mV, and passing in time, and bacterium forms microbial film at anode surface, and voltage slowly increases.After starting 5d, voltage quickenings that gather way is Exponential growth, until biofilm development is stablized.After inoculating about 8d, load voltage tends towards stability gradually, and this shows the start battery success;
After the 9th step, electrolyzer start successfully, the access power supply, regulating input voltage is 0.2～1.2 volt, this moment, electrolyzer was in the hydrogen pattern of producing.H at cathode side
2Outlet is collected H by gas collector
2Gas is at the CO of anode side
2Pneumatic outlet is collected CO by gas collector
2Gas, after substrate consumption was complete, the waste liquid of anolyte compartment and cathode compartment was discharged by leakage fluid dram 4 and leakage fluid dram 6 respectively.
Conclusion: collected H
2Purity is high, can reach more than 90%, has reduced the process of later stage gas delivery, purifying, has reduced production cost.
The 4th goes on foot, pretreated cellulose matrix and enzyme liquid is mixed according to 100:0.8, add again entry, cellulose matrix is 1:5～1:8 with the ratio of amount of water, matrix is in a liquid state, and carry out sterilising treatment, the mixed solution of the cellulose matrix after the sterilization and enzyme liquid and water is added the anolyte compartment of electrolyzer by the opening for feed 3 of anolyte compartment; The mixed solution of the cellulose matrix that adds and enzyme liquid and water accounts for 15% of electrolytic cell anode chamber volume; Described enzyme liquid is cellulase, beta-glucosidase, zytase in 28mg: the 0.06mL:20mg ratio is mixed gained;
The 5th step, the opening for feed 3 of anaerobically fermenting bacterium by the anolyte compartment added the anolyte compartment of electrolyzers, inoculum size is 25% (v/v);
The 6th step, the opening for feed by the anolyte compartment 3 add the combination medium liquids, add-on by adding matrix 10%, the prescription that wherein makes up substratum is: 5.8gL
-1Na
2HPO
4, 3.2gL
-1KH
2PO
4, 0.1gL
-1CO (NH
2)
2, 0.7gL
-1NaCl, 0.24gL
-1MgSO
47H
2O, 16mgL
-1CaCl
22H
2O and 8mgL
-1ZnSO
47H
2The solution of O;
The 7th step, be 45mML with concentration
-1Potassium sulfite solution, the opening for feed 5 by cathode compartment adds cathode compartments, and to add simultaneously pH be that 4.8 concentration are 60mML
-1The phosphoric acid salt sustained release dosage, the phosphoric acid salt sustained release dosage that adds and the ratio of potassium sulfite solution are 4:1; The volume of the solution that adds at cathode compartment equates to get final product with the volume of the added solution in anolyte compartment.Realize the coupling that the liquid synchronously enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell produce hydrogen.
Embodiment 4 is as different from Example 2:
The 4th goes on foot, pretreated cellulose matrix and enzyme liquid is mixed according to 100:0.8, add again entry, cellulose matrix is 1:1～1:4 with the ratio of amount of water, it is solid-state that mixed solution is, the anolyte compartment that the mixed solution of cellulose matrix and enzyme liquid and water is added electrolyzer by the opening for feed 3 of anolyte compartment, the volume of the mixed solution of the cellulose matrix that adds and enzyme liquid and water is 15% of electrolytic cell anode chamber volume, and described enzyme liquid is cellulase, beta-glucosidase, zytase in 30mg: the 0.04mL:22mg ratio is mixed gained.
The 7th step, be 45mML with concentration
-1Potassium sulfite solution, opening for feed 5 by cathode compartment adds cathode compartment, the opening for feed 5 of anaerobically fermenting bacterium by cathode compartment added the cathode compartment of electrolyzers, and inoculum size is 15% (v/v), and the volume of the solution that adds at cathode compartment equates with the volume of the added solution in anolyte compartment; Realized that the solid-state synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell produce the coupling of hydrogen.
The mixed bacterium of the 6th step, anaerobically fermenting bacterium and photosynthetic bacterium, according to the inoculum size inoculation of 15%v/v, the combination medium liquid adds the electrolytic cell anode chamber from feeding mouth take speed as 100～200mL/min together.Nutrient solution is flowed out from leakage fluid dram with identical speed, make the matrix amount in the electrolyzer keep constant fermenting process.The prescription that wherein makes up substratum is: 6.2gL
-1Na
2HPO
4, 2.8gL
-1KH
2PO
4, 0.12gL
-1CO (NH
2)
2, 0.7gL
-1NaCl, 0.2gL
-1MgSO
47H
2O, 15mgL
-1CaCl
22H
2O and 12mgL
-1CoCl
26H
2The solution of O;
The 7th step, be 45mML with concentration
-1Potassium sulfite solution and pH be that 4.8 concentration are 50mML
-1The phosphoric acid salt sustained release dosage, add the cathode of electrolytic tank chamber take speed as 100～200mL/min from feeding mouth at the cathode compartment opening for feed; Nutrient solution is flowed out from leakage fluid dram with identical speed, make the matrix amount in the electrolyzer keep constant fermenting process;
The 8th step, electrolyzer external load resistance start electrolyzer.The electrolyzer initial start stage is 0～4d, and the load both end voltage is about 0mV, and passing in time, and bacterium forms microbial film at anode surface, and voltage slowly increases.After starting 5～8d, voltage quickenings that gather way is Exponential growth, until biofilm development is stablized.Behind inoculation 7～10d, load voltage tends towards stability gradually, and this shows the start battery success;
After the 9th step, electrolyzer start successfully, the access power supply, regulating input voltage is 0.2～1.2 volt, this moment, electrolyzer was in the hydrogen pattern of producing.At H
2The collection and confinement of gases outlet is collected H by gas collector
2Gas is at CO
2The collection and confinement of gases outlet is collected CO by gas collector
2Gas.Realize that the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose continous way and microorganism electrolysis cell produce the coupling of hydrogen.
The 4th step, pretreated cellulose matrix and enzyme liquid are mixed according to 100:0.9, and carry out sterilising treatment, described enzyme liquid is that cellulase, beta-glucosidase, zytase mix gained in the 36mg:0.05mL:24mg ratio;
The 5th step, the mixed solution of cellulose matrix and enzyme liquid and anaerobically fermenting bacterium are inoculated according to the inoculum size of 15% (v/v);
The 6th step, the mixed solution of postvaccinal cellulose matrix and enzyme liquid is added the electrolytic cell anode chambers from feeding mouth 3 with the combination medium liquid, the add-on that makes up medium liquid by adding matrix 30%, the prescription that wherein makes up substratum is: 5.8gL
-1Na
2HPO
4, 3.1gL
-1KH
2PO
4, 0.1gL
-1CO (NH
2)
2, 0.6gL
-1NaCl, 0.24gL
-1MgSO
47H
2O, 16mgL
-1CaCl
22H
2O and 10mgL
-1CuCl
22H
2The solution of O;
The 7th step, be 50mML with concentration
-1The opening for feed 5 of potassium ferricyanide solution by cathode compartment add cathode compartments, and to add simultaneously pH be that 4.8 concentration are 60mML
-1The phosphoric acid salt sustained release dosage, the phosphoric acid salt sustained release dosage that adds and the ratio of potassium ferricyanide solution are 4:1.The opening for feed 5 of photosynthetic bacterium by cathode compartment added the cathode compartment of electrolyzers, and inoculum size is 15% (v/v), and the volume of the solution that adds at cathode compartment equates to get final product with the volume of the added solution in anolyte compartment;
The 8th step, electrolyzer external load resistance start electrolyzer.The electrolyzer initial start stage is 2d, and the load both end voltage is about 0mV, and passing in time, and bacterium forms microbial film at anode surface, and voltage slowly increases.After starting 5d, voltage quickenings that gather way is Exponential growth, until biofilm development is stablized.After inoculating about 7d, load voltage tends towards stability gradually, and this shows the start battery success;
After the 9th step, electrolyzer start successfully, the access power supply, regulating input voltage is 0.2～1.2 volt, this moment, electrolyzer was in the hydrogen pattern of producing.H at cathode side
2Outlet is collected H by gas collector
2Gas is at the CO of anode side
2Pneumatic outlet is collected CO by gas collector
2Gas.After substrate consumption was complete, the waste liquid of anolyte compartment and cathode compartment was discharged by leakage fluid dram 4 and leakage fluid dram 6 respectively.Again fill the materials such as fresh matrix and substratum, be implemented in the synchronous enzymatic hydrolysis and fermentation of microorganism electrolysis cell system Mierocrystalline cellulose sequence batch (and produce hydrogen.
Conclusion: by the coupling of the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen, so that collected H
2Purity is high, can reach more than 90%, has reduced the process of later stage gas delivery, purifying, has reduced production cost.
Claims (6)
1. based on the coupling process of the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen, it is characterized in that, comprise the steps:
A. incite somebody to action not pre-treatment or pretreated cellulose matrix and make up enzyme liquid according to 100:(0.5～1) mix, and with in the electrolyzer after it adding sterilising treatment, add-on is 10%～20% of electrolyzer volume;
B. anaerobically fermenting bacterium, photosynthetic bacterium or the mixed bacterium of the two are added electrolyzer, inoculum size is 5%～35% (v/v);
C. in electrolyzer, add the combination medium liquid, add-on by adding matrix 10%～35%;
D. the electrolyzer outer meeting resistance starts electrolyzer, after electrolyzer starts successfully, and the access external power source, regulating input voltage is 0.2～1.2 volt, electrolyzer is in the hydrogen pattern of producing; At H
2The collection and confinement of gases outlet is collected H by gas collector
2Gas is at CO
2The collection and confinement of gases outlet is collected CO by gas collector
2Gas, treat that substrate consumption is complete after, waste liquid is discharged by leakage fluid dram 4.
2. the coupling process that produces hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell according to claim 1, it is characterized in that: when microorganism electrolysis cell adopts the double-chamber microbiological electrolyzer, in steps A, the anolyte compartment of material that B, C add for the adding electrolyzer, and before step D, also carry out following steps:
Be 20～60mML with concentration
-1Potassium ferricyanide solution or concentration be 20～60mML
-1Potassium sulfite solution add in the cathode compartment of electrolyzer, and equate with the volume of the added solution in anolyte compartment at the volume of solution that cathode compartment adds.
3. the coupling process that produces hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell according to claim 2, it is characterized in that: when microorganism electrolysis cell adopts the double-chamber microbiological electrolyzer, the mixed bacterium or single photosynthetic bacterium or the anaerobically fermenting bacterium that also add simultaneously anaerobically fermenting bacterium and photosynthetic bacterium in the cathode of electrolytic tank chamber, inoculum size is 5%～35% (v/v), and equates with the volume of the added solution in anolyte compartment at the volume of solution that cathode compartment adds.
4. the coupling process that produces hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell according to claim 3, it is characterized in that: when microorganism electrolysis cell adopts the double-chamber microbiological electrolyzer, in cathode compartment, also add pH and be 4.8, concentration is 40～80mML
-1The phosphoric acid salt sustained release dosage, the ratio of the phosphoric acid salt sustained release dosage that adds and potassium ferricyanide solution or potassium sulfite solution is 4:1; And the volume at solution that cathode compartment adds equates with the volume of the added solution in anolyte compartment.
5. according to claim 1,2,3 or 4 described coupling process based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell product hydrogen, it is characterized in that: described combination enzyme liquid is cellulase, beta-glucosidase, zytase, mixes in 22～40mg: 0.02～0.08mL:10～28mg ratio.
6. according to claim 5ly produce the coupling process of hydrogen based on the synchronous enzymatic hydrolysis and fermentation of Mierocrystalline cellulose and microorganism electrolysis cell, prescription of its combination substratum is: 5.8～6.2gL
-1Na
2HPO
4, 2.8～3.2gL
-1KH
2PO
4, 0.08～0.12gL
-1CO (NH
2)
2, 0.5～0.7gL
-1NaCl, 0.2～0.24gL
-1MgSO
47H
2O, 14～16mgL
-1CaCl
22H
2O and 6～12mgL
-1Trace element solution, wherein trace element is: Mo, Fe, Zn, Co or Cu.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103523866A (en) * | 2013-09-17 | 2014-01-22 | 吉林大学 | Biological cathode method of degrading organic matters and recycling energy contained in organic matters |
| CN104340957A (en) * | 2013-07-29 | 2015-02-11 | 中国科学院大连化学物理研究所 | Method for preparing hydrogen through photocatalytic decomposition of water by virtue of photosystem II and semiconductor hybrid system |
| CN104961240A (en) * | 2015-05-26 | 2015-10-07 | 中国科学院过程工程研究所 | Waste leachate treatment method and microbial electrolytic battery |
| CN109628496A (en) * | 2018-12-21 | 2019-04-16 | 广西大学 | A method of the collaboration based on microorganism electrolysis cell utilizes thermophilic flora high efficiente callback hydrogen from cellulose |
| CN113355365A (en) * | 2021-07-28 | 2021-09-07 | 宜宾学院 | Hydrogen production method using phyllostachys pubescens as raw material |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1856706A (en) * | 2003-07-10 | 2006-11-01 | 斯蒂奇威特苏斯可持续水技术中心 | Bio-electrochemical process for producing hydrogen |
| CN101459254A (en) * | 2008-12-29 | 2009-06-17 | 天津理工大学 | Method and apparatus for electricity production by stalk cellulose microbial degradation |
| CN101760479A (en) * | 2008-12-25 | 2010-06-30 | 中国科学院成都生物研究所 | Method for producing hydrogen through biomass fermentation and photosynthetic coupling |
| US20120082869A1 (en) * | 2009-04-07 | 2012-04-05 | Arizona Board Of Regents | Microbial Electrolytic Cell |
-
2012
- 2012-09-28 CN CN201210369997.9A patent/CN102876724B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1856706A (en) * | 2003-07-10 | 2006-11-01 | 斯蒂奇威特苏斯可持续水技术中心 | Bio-electrochemical process for producing hydrogen |
| CN101760479A (en) * | 2008-12-25 | 2010-06-30 | 中国科学院成都生物研究所 | Method for producing hydrogen through biomass fermentation and photosynthetic coupling |
| CN101459254A (en) * | 2008-12-29 | 2009-06-17 | 天津理工大学 | Method and apparatus for electricity production by stalk cellulose microbial degradation |
| US20120082869A1 (en) * | 2009-04-07 | 2012-04-05 | Arizona Board Of Regents | Microbial Electrolytic Cell |
Non-Patent Citations (1)
| Title |
|---|
| 廖强 等: "光合细菌生物制氢反应器研究进展", 《应用与环境生物学报》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104340957A (en) * | 2013-07-29 | 2015-02-11 | 中国科学院大连化学物理研究所 | Method for preparing hydrogen through photocatalytic decomposition of water by virtue of photosystem II and semiconductor hybrid system |
| CN104340957B (en) * | 2013-07-29 | 2017-03-15 | 中国科学院大连化学物理研究所 | Photosynthetical system two and the method for semiconductor hybrid system photocatalytic hydrogen production by water decomposition gas |
| CN103523866A (en) * | 2013-09-17 | 2014-01-22 | 吉林大学 | Biological cathode method of degrading organic matters and recycling energy contained in organic matters |
| CN103523866B (en) * | 2013-09-17 | 2015-12-23 | 吉林大学 | A kind of organic biological-cathode method reclaiming its contained energy of degrading |
| CN104961240A (en) * | 2015-05-26 | 2015-10-07 | 中国科学院过程工程研究所 | Waste leachate treatment method and microbial electrolytic battery |
| CN109628496A (en) * | 2018-12-21 | 2019-04-16 | 广西大学 | A method of the collaboration based on microorganism electrolysis cell utilizes thermophilic flora high efficiente callback hydrogen from cellulose |
| CN109628496B (en) * | 2018-12-21 | 2022-05-03 | 广西大学 | Method for efficiently recovering hydrogen from cellulose by synergistically utilizing thermophilic floras based on microbial electrolysis cell |
| CN113355365A (en) * | 2021-07-28 | 2021-09-07 | 宜宾学院 | Hydrogen production method using phyllostachys pubescens as raw material |
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