CN115125274A - Method for producing bio-based chemicals based on microbial ecological potential regulation and control anaerobic fermentation system - Google Patents

Method for producing bio-based chemicals based on microbial ecological potential regulation and control anaerobic fermentation system Download PDF

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CN115125274A
CN115125274A CN202210788209.3A CN202210788209A CN115125274A CN 115125274 A CN115125274 A CN 115125274A CN 202210788209 A CN202210788209 A CN 202210788209A CN 115125274 A CN115125274 A CN 115125274A
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于佳动
赵立欣
安柯萌
姚宗路
罗娟
申瑞霞
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Institute of Environment and Sustainable Development in Agriculturem of CAAS
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Abstract

The invention belongs to the technical field of anaerobic fermentation, and provides a method for producing bio-based chemicals based on a microorganism ecological niche regulation and control anaerobic fermentation system. The method comprises the steps of crushing straws, carbon chain extension reaction, hydrolysis acidification to produce short acid, methanation of acid to produce methane, discharging and the like. The invention achieves the aim of directionally synthesizing the product by regulating the high-solid-content anaerobic fermentation system through the microbial ecological niche, enhances the cooperative metabolic mechanism between acid-producing bacteria and methanogenic bacteria, improves the activity of microbial communities with different functions, realizes the co-production of medium-chain fatty acid, short-chain fatty acid and methane in the high-solid-content anaerobic fermentation system for the first time, further widens the energy utilization way of the straw and improves the added value of the product.

Description

Method for producing bio-based chemicals based on microbial ecological potential regulation and control anaerobic fermentation system
Technical Field
The invention relates to the technical field of anaerobic fermentation, in particular to a method for producing bio-based chemicals based on a microorganism ecological level regulation and control anaerobic fermentation system.
Background
It is reported that a large number of planting wastes such as cereal straws (rice straw, wheat straw, corn straw, etc.), bean straws, potato straws, oil plant straws, and various fruits and vegetables straws are generated every year due to agricultural planting. Anaerobic fermentation is one of agricultural waste resource utilization technologies, a high solid content anaerobic fermentation device has the advantage of large unit volume raw material treatment capacity, but the straw contains a large amount of cellulose and hemicellulose, so that the problems of low starting speed of high solid content fermentation, low substance conversion efficiency, low gas production rate and the like are caused, the utilization way of the produced biogas is limited, the self additional value is low, the commercial application of biogas residue and biogas liquid fertilizer is not realized, and the technical route of anaerobic fermentation gas fertilizer co-production hardly achieves the effect of producing high additional value products. Therefore, it is urgently needed to provide a continuous, stable, efficient and high-value anaerobic fermentation process regulation and control method.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a method for producing bio-based chemicals based on a microorganism ecological niche regulation and control anaerobic fermentation system, which is characterized in that a process regulation and control method is designed by utilizing the difference of metabolic characteristics of microorganisms with different functions in different stages of continuous anaerobic fermentation of straws with high solid content, the interspecific action relationship of the microorganisms is changed in different fermentation stages, the directional metabolic rate of the microorganisms is enhanced, and the integrated production of the bio-based chemicals is realized.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for producing bio-based chemicals based on a microbial ecological niche regulation and control anaerobic fermentation system, which comprises the following steps:
(1) mixing the straw winding mass with whey wastewater to obtain a mixed material;
(2) mixing the mixed material with anaerobic granular sludge, adding carbon chain extension functional microorganism bacteria, and performing first-stage fermentation to obtain a first fermentation product;
(3) mixing the first fermentation product with cellulose decomposition bacteria, and performing second-stage fermentation to obtain a second fermentation product;
(4) mixing the second fermentation product with cellulase, and performing third-stage fermentation to obtain a third fermentation product;
the first fermentation product comprises medium chain fatty acids;
the second fermentation product comprises short chain fatty acids;
the third fermentation product comprises methane and CO 2 。
Preferably, in the step (1), the particle size of the straw winding group is 1-3 cm, the content of lactic acid in whey wastewater is 3-5 g/L, the using amount ratio of the straw winding group to the whey wastewater is 5-10 g:1mL, and the solid content of the mixed material is 15-25%.
Preferably, in the step (2), the addition amount of the anaerobic granular sludge is 20-40% of the mass of the mixed material, the granularity of the anaerobic granular sludge is 60-70%, the particle diameter of the anaerobic granular sludge is 0.5-4 mm, the water content of the anaerobic granular sludge is 85-95%, the inoculation amount of the carbon chain extension functional microbial bacteria is 20-35% of the mass of the mixed material, and the carbon chain extension functional microbial bacteria comprise clostridium kluyveri, clostridium and megacoccus aegypti.
Preferably, in the step (2), the time of the first-stage fermentation is 5-10 d, the temperature of the first-stage fermentation is 35-45 ℃, and the pH value of the first-stage fermentation is 5-6.
Preferably, the inoculation amount of the cellulolytic bacteria in the step (3) is 10-20% of the mass of the first fermentation product, and the cellulolytic bacteria comprise bacteroides vulgatus and ruminal cellulolytic bacteria.
Preferably, the time of the second-stage fermentation in the step (3) is 4-6 d, the temperature of the second-stage fermentation is 45-55 ℃, and the pH value of the second-stage fermentation is 8-9.
Preferably, the adding amount of the cellulase in the step (4) is 1-5% of the mass of the second fermentation product, the enzyme activity of the cellulase is more than 400U/mg, and the cellulase comprises beta-1, 4-glucan hydrolase, beta-glucan cellobiohydrolase or beta-glucosidase.
Preferably, the time of the third-stage fermentation in the step (4) is 8-12 d, the temperature of the third-stage fermentation is 35-45 ℃, and the pH value of the third-stage fermentation is 6-8.
Preferably, the feeding amount and the discharging amount of the first-stage fermentation in the step (2), the second-stage fermentation in the step (3) and the third-stage fermentation in the step (4) are the same.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention provides a continuous, stable, efficient and high-value anaerobic fermentation process regulation method which is based on microbial ecological niche regulation and control of a high-solid-content anaerobic fermentation system to synchronously produce medium-chain fatty acid, short-chain fatty acid and methane and synchronously and efficiently degrade soluble substances, hemicellulose and cellulose of straws.
2. According to the invention, based on the regulation and control of microbial ecological niches, the environmental conditions of the survival of strains in each stage are regulated and controlled, the waste is treated in 3 stages, microbial floras with different functions are added, cellulase is added when the consumption of available substances is bottleneck, and finally, the decomposition of a substrate is stimulated, so that the straw degradation is more sufficient, and the effect of resource utilization of planting waste is achieved.
3. The invention achieves the purpose of directionally synthesizing the product by regulating and controlling the high-solid-content anaerobic fermentation system through the microbial ecological niche, enhances the cooperative metabolic mechanism between acid-producing bacteria and methanogenic bacteria, improves the activity of microbial communities with different functions, realizes the co-production of medium-chain fatty acid, short-chain fatty acid and methane in the high-solid-content anaerobic fermentation system for the first time, further widens the energy utilization way of the straw and improves the added value of the product.
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FIG. 1 is a high solids anaerobic fermentation system apparatus for the process of the present invention;
FIG. 2 is a reaction scheme of the process of the present invention.
Detailed Description
The invention provides a method for producing bio-based chemicals based on a microbial ecological niche regulation and control anaerobic fermentation system, which comprises the following steps:
(1) mixing the straw winding mass with whey wastewater to obtain a mixed material;
(2) mixing the mixed material with anaerobic granular sludge, adding a carbon chain extension functional microorganism strain, and performing first-stage fermentation to obtain a first fermentation product;
(3) mixing the first fermentation product with cellulose decomposition bacteria, and performing second-stage fermentation to obtain a second fermentation product;
(4) mixing the second fermentation product with cellulase, and performing third-stage fermentation to obtain a third fermentation product;
the first fermentation product comprises medium chain fatty acids;
the second fermentation product comprises short chain fatty acids;
the third fermentation product comprises methane and CO 2 。
In the invention, the particle size of the straw winding mass in the step (1) is preferably 1-3 cm, and further preferably 2 cm; the whey wastewater is preferably milk processing acid whey wastewater; the content of lactic acid in the whey wastewater is preferably 3-5 g/L, and more preferably 4 g/L; the dosage ratio of the straw winding mass to the whey wastewater is preferably 5-10 g:1mL, and more preferably 8g:1 mL; the solid content of the mixed material is preferably 15-25%, more preferably 18-22%, and even more preferably 20%.
In the invention, the adding amount of the anaerobic granular sludge in the step (2) is preferably 20-40% of the mass of the mixed material, more preferably 25-35% of the mass of the mixed material, and even more preferably 30% of the mass of the mixed material; the granularity of the anaerobic granular sludge is preferably 60-70%, and is further preferably 65%; the particle diameter of the anaerobic granular sludge is preferably 0.5-4 mm, and is further preferably 2 mm; the water content of the anaerobic granular sludge is preferably 85-95%, and more preferably 90%; the inoculation amount of the carbon chain extension function microbial bacteria is preferably 20-35% of the mass of the mixed material, more preferably 28-32% of the mass of the mixed material, and even more preferably 30% of the mass of the mixed material; the carbon chain extending functional microorganism preferably comprises Clostridium kluyveri, Clostridium and giant coccus aegypti.
In the invention, the time of the first-stage fermentation in the step (2) is preferably 5-10 d, and more preferably 9 d; the temperature of the first-stage fermentation is preferably 35-45 ℃, more preferably 38-42 ℃, and even more preferably 40 ℃; the pH value of the first stage fermentation is preferably 5-6, more preferably 5.2-5.8, and even more preferably 5.5. The first stage of fermentation aims to improve that lactic acid bacteria further produce lactic acid and acetic acid by utilizing straw soluble components, and carbon chain extension functional bacteria rapidly synthesize medium-chain fatty acid by utilizing the lactic acid and the acetic acid as substrates, so that functional complementation is formed among strains, and methanogens in the granular sludge cannot produce methane in a low pH environment and are in a functional dormant state. In addition, the carbon chain extension functional bacteria are added to strengthen the reverse beta oxidation process in the carbon chain extension metabolic pathway and promote the expression of key enzymes such as acetyl coenzyme A, butyryl coenzyme A and the like.
In the present invention, the amount of the cellulolytic bacteria inoculated in step (3) is preferably 10 to 20% by mass of the first fermentation product, more preferably 13 to 17% by mass of the first fermentation product, and even more preferably 15% by mass of the first fermentation product; the cellulolytic bacteria preferably include Bacteroides vulgatus and ruminolytic bacteria.
In the invention, the time of the second stage fermentation in the step (3) is preferably 4-6 d, and more preferably 5 d; the temperature of the second stage fermentation is preferably 45-55 ℃, and further preferably 50 ℃; the pH value of the second stage fermentation is preferably 8-9, and further preferably 8.5. The purpose of the second stage fermentation is to mainly produce short-chain fatty acid by utilizing the residual soluble components, hemicellulose and cellulose with relatively loose structure of the raw materials. And in the second-stage fermentation process, biogas slurry is refluxed every day, the biogas slurry is from liquid obtained after solid-liquid separation at a discharge port, the reflux ratio is set by adopting a method for monitoring the accumulation amount of volatile acid in the second-stage fermentation by sampling, when the concentration of the volatile acid is less than 2g/L, the reflux ratio in the methanogenesis stage is kept at 30-40%, preferably 35%, when the concentration of the volatile acid is more than 2g/L, the reflux ratio is increased to 40-50%, and the reflux ratio is further increased to 45%.
In the invention, the adding amount of the cellulase in the step (4) is preferably 1-5% of the mass of the second fermentation product, more preferably 2-4%, and even more preferably 3%; the enzyme activity of the cellulase is preferably more than 400U/mg; the cellulase preferably comprises a beta-1, 4-glucan cellobiohydrolase, a beta-glucan cellobiohydrolase, or a beta-glucosidase, and more preferably a beta-1, 4-glucan hydrolase.
In the invention, the time of the third stage fermentation in the step (4) is preferably 8-12 d, and more preferably 10 d; the temperature of the third stage fermentation is preferably 35-45 ℃, more preferably 38-42 ℃, and even more preferably 40 ℃; the pH value of the third stage fermentation is preferably 6-8, and more preferably 7. The third stage fermentation aims at utilizing the nondegradable hemicellulose, cellulose and a small amount of lignin in the raw materials to carry out fermentation post-treatment under the action of adding cellulase so as to fully release the methanogenesis potential of the raw materials.
In the present invention, the feeding amount and the discharging amount of the first-stage fermentation in the step (2), the second-stage fermentation in the step (3) and the third-stage fermentation in the step (4) are the same. The first stage fermentation, the second stage fermentation and the third stage fermentation belong to three stages of continuous fermentation, different strains are inoculated into the feeding ports (1), (2) and (3) in the figure 1 in each stage, materials are mixed in respective areas of each stage by a stirring device in the device, and the materials are integrally kept to uniformly move from the feeding end to the discharging end.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
The carbon chain extension functional microbial bacteria in the following embodiment are obtained by a high-throughput sequencing method from a continuous fermentation acid production reactor using straws as a substrate, wherein the microorganism is 25-35% of clostridium kluyveri, 15-20% of clostridium, 15-20% of megacoccus aegypti, 10-15% of vibrio succinogenes, 5-10% of bifidobacterium ruminal bacteria, 5-10% of lactic acid bacteria, 2-5% of bacillus aceticus and 10-15% of the rest; the cellulose decomposition bacteria are from straw high-temperature (55 ℃) compost, and through high-throughput sequencing, 25-35% of common bacteroides, 15-25% of rumen-inhabiting cellulose decomposing bacteria, 10-20% of acidophilic bacillus, 5-10% of cellulose bacillus, 5-10% of vibrio cellulosae and 10-15% of the rest; cellulase was purchased from Beijing Boototta technologies, Inc. under CAS number 9012-54-8.
Example 1
(1) The straw is pre-soaked by adding milk processing acid whey wastewater (wherein the dosage ratio of the straw winding group to the milk processing acid whey wastewater is 8g:1mL, and the content of lactic acid in the milk processing acid whey wastewater is 4g/L) into winding groups with the particle size of 2cm, feeding the materials until the solid content of the whey wastewater soaking the straw is 20%, and uniformly mixing the materials until the pH value is 5.5.
(2) Setting the hydraulic retention time of the high solid content fermentation system to be 25 d. And (2) simultaneously adding the straws obtained in the step (1) and anaerobic granular sludge with the inoculation ratio of 30% into a high-solid-content continuous fermentation device with the working volume of 150L from a feed inlet. And 6kg of materials are added into a feeding port every day, so that the normal operation of the continuous fermentation system with high solid content is maintained.
(3) The first stage fermentation time is 9 days, the fermentation temperature is kept at 40 ℃ by a heat preservation device, the pH value is controlled at 5.5, 30% of microorganism strain with carbon chain extending function (clostridium kluyveri: clostridium clostridia: megacoccus aegythii ═ 1:1:1) is added into a feeding port (1) at the upper part, meanwhile, a liquid sample containing medium-chain fatty acid is collected at a sampling port at the lower part of the device every day, and the yield of caproic acid reaches 10 g/L.
(4) The second stage fermentation is 10-15 days of hydraulic retention time, the fermentation temperature is kept at 50 ℃ by a heat preservation device, the fermentation time is 6 days, cellulose decomposition bacteria (common bacteroides: rumen decomposition cellulose bacteria 1:1) are added into a feeding port (2) at the upper part when the materials move to the second stage, the inoculation amount is 15 percent, and the pH value is adjusted to 8.5.
(5) And in the second stage fermentation process, biogas slurry reflux is carried out every day, and the biogas slurry is from the liquid obtained after solid-liquid separation at the discharge hole. The reflux ratio is set by adopting a method of sampling and monitoring the accumulation amount of the volatile acid in the 2 nd stage, when the concentration of the volatile acid is less than 2g/L, the reflux ratio in the methanogenesis stage is continuously kept at 35%, and when the concentration of the volatile acid is more than 2g/L, the reflux ratio is improved to 45%.
(6) The third stage of fermentation is 16-25 days of retention time, the fermentation temperature is kept at 40 ℃ by a heat preservation device, the fermentation time is 10 days, the pH value is not adjusted in the stage after the methane production is stable, the pH value can be kept neutral, when the material is operated to the third stage, cellulase (beta-1, 4-glucan hydrolase) is added when a feeding port (3) at the upper part is provided with a feeding port, the adding amount of the cellulase is 3% (500U/mg), the residual cellulose is decomposed, and the methane is produced.
(7) After fermenting for 25 days, the yield of the collected and generated biogas reaches 450m 3 The functional microorganism quantity of each stage is improved by 120 percent, and the cellulase activity is improved by 50 percent.
In conclusion, the invention utilizes the difference of metabolism of functional microorganisms in different fermentation stages to change the ecological niche of the microorganisms and dominate the biochemical reaction path to change to produce the target product, and the designed process regulation and control conditions are suitable for the growth of the microorganisms in three stages, are suitable for the mutual coordination and mutual promotion among the functional microorganisms, and obviously improve the yield of medium-chain fatty acids, short-chain fatty acids and methane in the high-solid-content anaerobic fermentation process.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and amendments can be made without departing from the principle of the present invention, and these modifications and amendments should also be considered as the protection scope of the present invention.

Claims (9)

1. A method for producing bio-based chemicals based on a microbial ecological niche regulation anaerobic fermentation system is characterized by comprising the following steps:
(1) mixing the straw winding mass with whey wastewater to obtain a mixed material;
(2) mixing the mixed material with anaerobic granular sludge, adding carbon chain extension functional microorganism bacteria, and performing first-stage fermentation to obtain a first fermentation product;
(3) mixing the first fermentation product with cellulose decomposing bacteria, and performing second-stage fermentation to obtain a second fermentation product;
(4) mixing the second fermentation product with cellulase, and performing third-stage fermentation to obtain a third fermentation product;
the first fermentation product comprises medium chain fatty acids;
the second fermentation product comprises short chain fatty acids;
the third fermentation product comprises methane and CO 2 。
2. The method for producing bio-based chemicals based on the microbial niche controlled anaerobic fermentation system according to claim 1, wherein the particle size of the straw winding mass in step (1) is 1-3 cm, the content of lactic acid in the whey wastewater is 3-5 g/L, the dosage ratio of the straw winding mass to the whey wastewater is 5-10 g:1mL, and the solid content of the mixed material is 15-25%.
3. The method for producing bio-based chemicals based on the microorganism niche-regulated anaerobic fermentation system according to claim 1 or 2, wherein the amount of the added anaerobic granular sludge in step (2) is 20-40% of the mass of the mixture, the granularity of the anaerobic granular sludge is 60-70%, the particle diameter of the anaerobic granular sludge is 0.5-4 mm, the water content of the anaerobic granular sludge is 85-95%, the inoculation amount of the carbon chain extension functional microorganism is 20-35% of the mass of the mixture, and the carbon chain extension functional microorganism comprises clostridium kluyveri, clostridium sp and megacoccus aegythii.
4. The method for producing bio-based chemicals based on the microbial niche controlled anaerobic fermentation system according to claim 3, wherein the time of the first stage fermentation in step (2) is 5-10 d, the temperature of the first stage fermentation is 35-45 ℃, and the pH value of the first stage fermentation is 5-6.
5. The method for producing bio-based chemicals based on the microbial niche-regulated anaerobic fermentation system according to claim 4, wherein the cellulose-decomposing bacteria in step (3) are inoculated in an amount of 10-20% of the first fermentation product, and the cellulose-decomposing bacteria comprise Bacteroides vulgatus and Rumex ruminalis.
6. The method for producing bio-based chemicals based on the microbial niche controlled anaerobic fermentation system according to claim 5, wherein the time of the second stage fermentation in step (3) is 4-6 d, the temperature of the second stage fermentation is 45-55 ℃, and the pH value of the second stage fermentation is 8-9.
7. The method for producing bio-based chemicals based on the microbial niche regulated anaerobic fermentation system of claim 6, wherein the cellulase added in step (4) is 1-5% of the mass of the second fermentation product, the cellulase has an enzymatic activity of > 400U/mg, and the cellulase comprises β -1, 4-glucan hydrolase, β -glucan cellobiohydrolase or β -glucosidase.
8. The method for producing bio-based chemicals based on the microbial niche controlled anaerobic fermentation system of claim 7, wherein the time of the third stage fermentation in step (4) is 8-12 d, the temperature of the third stage fermentation is 35-45 ℃, and the pH value of the third stage fermentation is 6-8.
9. The method for producing bio-based chemicals by using a microorganism niche-based controlled anaerobic fermentation system according to claim 7, wherein the feeding amount and the discharging amount of the first-stage fermentation in step (2), the second-stage fermentation in step (3) and the third-stage fermentation in step (4) are the same.
CN202210788209.3A 2022-07-06 2022-07-06 Method for producing bio-based chemicals based on microbial ecological potential regulation and control anaerobic fermentation system Pending CN115125274A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115354064A (en) * 2022-10-21 2022-11-18 中国农业科学院农业环境与可持续发展研究所 Method for producing medium-chain fatty acid by two-phase partition of anaerobic dry fermentation

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115354064A (en) * 2022-10-21 2022-11-18 中国农业科学院农业环境与可持续发展研究所 Method for producing medium-chain fatty acid by two-phase partition of anaerobic dry fermentation

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