CN102373238B - Bacillus alcalophilus and application thereof to fermentative hydrogen production - Google Patents
Bacillus alcalophilus and application thereof to fermentative hydrogen production Download PDFInfo
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- CN102373238B CN102373238B CN2011103418330A CN201110341833A CN102373238B CN 102373238 B CN102373238 B CN 102373238B CN 2011103418330 A CN2011103418330 A CN 2011103418330A CN 201110341833 A CN201110341833 A CN 201110341833A CN 102373238 B CN102373238 B CN 102373238B
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Abstract
The invention relates to bacillus alcalophilus and an application thereof to fermentative hydrogen production. The steps of sludge and/or organic waste pretreatment and fermentative hydrogen production are adopted. The bacillus alcalophilus is characterized in that in the sludge and/or organic waste pretreatment step, bacillus alcalophilus liquid is inoculated in sludge and/or organic wastes, and bacillus alcalophilus hydrolysis pretreatment sludge and/or organic wastes are obtained after the treatment. In addition, the prepared bacillus alcalophilus hydrolysis pretreatment sludge can also be used as hydrogen production floras to be inoculated into the organic wastes for producing hydrogen through fermentation. A bacillus alcalophilus sludge pretreatment method can be effectively used for screening efficient hydrogen production floras in the sludge, inhabiting the growth of hydrogen consumption bacteria and killing pathogenic bacteria in the sludge and/or organic wastes. The sludge and/or organic wastes through all bacterial strain treatment has/have the large hydrogen generation quantity in the subsequent fermentative hydrogen production step, the purity of hydrogen gas is high, the delay time of the hydrogen production is short, and obvious economic values and development potential are realized.
Description
Technical field
The invention belongs to the fermentation and hydrogen production technical field, be specifically related to a kind of bacillus acidocldarius and the application in fermentation and hydrogen production thereof, be about to bacillus acidocldarius and be applied to mud/or the hydrolysis pre-treatment of organic waste, the mud after the recycling hydrolysis/or fermentation of organic wastes produce hydrogen; Also comprise a kind of bacillus acidocldarius A11 bacterial strain filtered out simultaneously.
Background technology
Due to coal, a large amount of uses of the Nonrenewable resources such as oil, the mankind not only are faced with the exhaustion of resource, but also can, to environment, produce Greenhouse effect.And Hydrogen Energy has become the most attractive substitute energy of 21st century due to its cleanliness without any pollution, high heating value, high thermal efficiency, the plurality of advantages such as applied widely, renewable.The generation of hydrogen at present can be passed through two kinds of approach: physico-chemical process and biological method.Physico-chemical process comprises water electrolysis method and water-gas method, but above-mentioned two kinds of methods are all that to consume electric energy or Nonrenewable energy resources be that cost obtains hydrogen.And biological method is to utilize the microorganism that produces hydrogen to come decomposing organic matter matter to produce hydrogen, it is a kind of technology of energy-conserving and environment-protective.
In the bio-hydrogen method, increase the quantity of producing the hydrogen microorganism and produce hydrogen activity, the hydrogenogens group of breeding high-efficiency, be to improve the important channel that the ferment for hydrogen production system is produced hydrogen usefulness.With utilize single bacterial strain to produce hydrogen to compare, utilize mixed culture to carry out anaerobe hydrogen manufacturing and there is obvious advantage, this is due to the synergy between bacterial classification, cause the hydrogen production potential of pure bacterium not as mixed strains, high product hydrogen microbial diversity is conducive to increase the hydrogen production potential of system for producing hydrogen, and wherein mud has maximum hydrogen production potential.Therefore, the stability and the hydrogen generation efficiency that in order to improve hydrogen, produce, the product hydrogen biological community structure of optimizing in mud is an important job.Simultaneously, eliminate the hydrogen-consuming bacteria groups such as methanogen floras, homoacetogenesis flora and sulfate reduction flora who exists in seed sludge, accelerate the sludge acclimatization process of reactive system, improve and produce hydrogen usefulness, process aerobic activated sludge and anaerobic activated sludge with the different pretreatments method, and it is just extremely important to start the ferment for hydrogen production systematic research.In recent years, the main hydrogenogens group who uses in physico-chemical process screening mud, hydrolyzing sludge or organic waste carry out fermentation and hydrogen production simultaneously, as thermal treatment (heating or sterilizing), chemical treatment (alkaline purification, acid treatment, add methane inhibitor etc.), machinery cracks methods such as (ultrasonic wave, microwave, freeze thawing etc.).
The excess sludge that has at present had some to utilize sewage work produces the technology report of hydrogen, the patent that the patent No. is ZL200310116142.6 discloses " a kind of method and apparatus that utilizes the hydrogen manufacturing of sewage plant excess sludge anaerobically fermenting ", it is that application alkali pretreating sludge produces hydrogen, pH is transferred to 12～13, and suppresses methanobacteria.But the strong basicity environment is very unfavorable to the subsequent biological treatment of mud, the landfill of excess sludge is larger to the damage ratio primary sludge of soil, and exogenously added alkali, increases the production run cost.The Patent Application Publication that application number is 200510066428.7 a kind of " by the thermal treatment of sewage treatment plant residual mud hydrogen manufacturing-fermentation and hydrogen production method ", its uses thermal treatment mud to produce hydrogen, at 121 ℃, 1.5 standard atmospheric pressures of pressure, process 0.5 hour.But this method time of lag long (from starting fermentation to producing hydrogen during this period of time), hydrogen purity is low, and power consumption is large, the manufacture of equipment is required high, increases production cost.And there are trichloromethane by product, mailland reaction inhibition anaerobic digestion, capital construction and the problems such as working cost is large, energy consumption is high, potential safety hazard of producing in the physico-chemical processes such as ultrasonic wave, ozone oxidation, oxychlorination, thermal hydrolysis, acid or alkaline purification.Therefore, just need to set up a kind of effective biological process preconditioning technique, to mud/or organic waste be hydrolyzed and utilize the method for its fermentation and hydrogen production.Simultaneously, filter out the required high-effective microorganism bacterial strain of biological process pre-treatment and also become mud/or the study hotspot in organic waste hydrolysis and fermentation and hydrogen production field.
Summary of the invention
The purpose of this invention is to provide a kind of bacillus acidocldarius and the application in fermentation and hydrogen production thereof, utilize bacillus acidocldarius to carry out pretreating sludge/or organic waste, hydrolyzing sludge/or the macromolecular structure of organic waste, the enrichment hydrogenogens, improve the efficiency of follow-up product hydrogen, to make up the deficiencies in the prior art.
The present invention relates to bacillus acidocldarius and produce the application in hydrogen in sludge fermentation, comprise sludge pretreatment and ferment for hydrogen production step, it is characterized in that above-mentioned pre-treatment step is inoculation bacillus acidocldarius bacterium liquid in mud, at 50～70 ℃ of lower pretreating sludge 2～24h, obtain bacillus acidocldarius pre-treatment hydrolysis mud, then proceed to the ferment for hydrogen production step.
Above-mentioned ferment for hydrogen production step is that bacillus acidocldarius pre-treatment hydrolysis mud is placed in to the ferment for hydrogen production reactor of sealing, and to nitrogen blowing 2～15min in mud, then sealing and fermenting produces hydrogen, and leavening temperature is 25～45 ℃.
Another aspect of the present invention relates to bacillus acidocldarius pre-treatment hydrolysis mud, is bacillus acidocldarius pre-treatment hydrolysis mud is inoculated in organic waste and carries out fermentation and hydrogen production as hydrogenogens group inoculum.
Above-mentioned organic waste also can first carry out pre-treatment with bacillus acidocldarius.
The volume ratio of above-mentioned mud or organic waste and bacillus acidocldarius bacterium liquid is 100: 3～15.
Above-mentioned mud can be excess sludge, river/seabed mud or the cultivating pool mud of sewage work.
Wherein bacillus acidocldarius is the bacillus acidocldarius Bacillus thermophilic A11 bacterial strain of screening.
Bacillus acidocldarius A11 (Bacillus thermophilic), deposit number is: CGMCC No.5309, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, postcode 100101, preservation date is on September 29th, 2011.
Bacillus acidocldarius sludge pre-treatment method of the present invention can screen the highly effective hydrogen yield flora in mud effectively, suppress the growth of hydrogen-consuming bacteria, destroy mud/or the macromolecular structure of organic waste simultaneously, improve mud/or the biology of organic waste degradation capability again, the reduce mud amount, kill mud/or organic waste in pathogenic bacterium.The pretreated mud of process A11 bacterial strain of the present invention/or organic waste, in follow-up ferment for hydrogen production step, the generation of hydrogen is large, and hydrogen purity is high, and the time of lag of producing hydrogen is short, has obvious economic worth and exploitation potential.
Embodiment
Ultimate principle of the present invention is as follows: mud/or organic waste in inoculate bacillus acidocldarius, under hot conditions, bacillus acidocldarius can be secreted extracellular enzyme, extracellular enzyme can destroy microorganism wall in mud, hydrolyzing sludge/or the macromolecular structure of organic waste, improve mud/or the biodegradability of organic waste; Extracellular enzyme and hot conditions can suppress or kill the hydrogen-consuming bacterias such as methanogen in mud simultaneously, and hydrogenogens can form spore at extreme environment, under suitable condition, can bring back to life, mud, after bacillus acidocldarius is processed, just can filter out highly effective hydrogen yield flora wherein.Therefore, mud/or organic waste after bacillus acidocldarius is processed, can be used for fermentation and hydrogen production.
Contrast mud is processed secondary fermentation product hydrogen through 65 ℃ of thermal treatments and bacillus acidocldarius, and it is 27.5ml H that the maximum of the rear mud of bacillus acidocldarius processing is produced the hydrogen rate
2/ g VS, produce the hydrogen rate and exceed 65 ℃ of corresponding thermal treatment mud 52.3%, in the gas of generation, only has H
2And CO
2, without methane, produce.Result shows, mud is after bacillus acidocldarius is processed, and hydrogenogens group activity is high, can reach and produce preferably the hydrogen effect.
Below in conjunction with specific embodiment, bacterial strain of the present invention and application thereof are described in detail
One, the screening of bacterial strain and physico-chemical property
1, the screening of bacterial strain:
Get green belt garden, Qingdao City soil and be placed in Erlenmeyer flask, 121 ℃ of high-temperature sterilizations, then cultivate 1～2 day in 65 ℃ of water-baths concussion, and what survive is thermophile bacteria, then adopt the thermophile bacteria of plate streak separation and concentration.Used medium is general beef-protein medium, and main component is as follows: extractum carnis 5g, and peptone 10g, agar 20g, NaCl 5g, water 1000ml, pH 7.0,1.05kg/cm
2, under 121 ℃ of conditions, sterilizing is 20 minutes.
2, the physico-chemical property of bacterial strain
The bacterium colony filtered out is light grey, translucent, shallow splitting, edge is irregular, convexity, this bacterial strain is gram-positive microorganism, thin rod shape, produce gemma, aerobic or facultative anaerobe, the catalase reaction is positive, but secreting amylase and proteolytic enzyme, optimum pH is 6.8～7.5, optimum growth temp is 65 ℃.
Two, the application of bacterial strain
The thermophile bacteria A11 that gets separation is placed in sewage plant excess sludge, the volume ratio of mud and thermophile bacteria bacterium liquid is 100: 3～100: 15, at 45～75 ℃ of lower lignin-sludge 2～24h, then get the ferment for hydrogen production reactor that the thermophile bacteria hydrolyzing sludge is placed in sealing, nitrogen blowing 2～15min, seal 25 ℃～45 ℃ of controlled fermentation temperature wherein, carry out fermentation and hydrogen production, produce hydrogen after 2～10h.
Hydrolysis and fermentation and hydrogen production for other organic waste, concrete grammar is as follows: the thermophile bacteria A11 that gets separation is placed in organic waste, the volume ratio of organic waste and thermophile bacteria bacterium liquid is 100: 3～100: 15, process 2～48h under 45～75 ℃, then inoculate the mud after thermophile bacteria is processed, be placed in the ferment for hydrogen production reactor of sealing, the ratio of mud and organic waste is 100: 5～100: 20, then to nitrogen blowing 2～15min in the ferment for hydrogen production reactor, sealing, 25 ℃～45 ℃ of controlled fermentation temperature, carry out fermentation and hydrogen production, produce hydrogen after 2～10h.
Embodiment 1:
Get municipal wastewater treatment plant second pond mud 100ml, the TSS of mud is 13.26g/L, and VSS is 7.41g/L.Be placed in the thermophile bacteria hydrolysis reactor, the bacterium liquid 10ml of inoculation thermophile bacteria Bacillus thermophilic A11 bacterial strain (gets the A11 inoculation in liquid nutrient medium, the bacterium liquid that 24h makes the A11 bacterial strain is cultivated in 65 ℃ of water-bath concussions, the substratum main component is: extractum carnis 5g, peptone 10g, NaCl 5g, water 1000ml, pH 7.0,1.05kg/cm
2, under 121 ℃ of conditions, sterilizing is 20 minutes), experimental temperature is set as 65 ℃, air flow is 0.21L/min, processes 48h, to discharge the organic nutrient substance in mud, hydrogenogens group in simultaneously again can enrichment mud, kill hydrogen-consuming bacteria and pathogenic bacterium, and can discharge the part heavy metal adsorbed in mud.After pre-treatment, the dissolution rate that the dissolution rate of TSS reaches 41.21%, VSS reaches 55.27%, and the nutrition in mud is released, and wherein the content of primary sludge protein rises to 1011mg/L from 87.13mg/L; The content of carbohydrate rises to 253mg/L from 10.45mg/L; SCOD (solvability COD), by the 245mg/L of primary sludge, brings up to 6342mg/L.
Then thermophile bacteria is hydrolyzed to pretreated mud and is put in serum bottle, control 35 ℃ of temperature, nitrogen blowing 5min, to remove the airborne oxygen in excess sludge upper strata, keep the anaerobic environment of fermentation and hydrogen production reactor, starts to produce hydrogen after 3 hours.Maximum product hydrogen rate is 27.5ml/gVS.Wherein the content of hydrogen is more than 80%, and all the other are carbonic acid gas, there is no the generation of methane gas.Produce the hydrogen rate and exceed 52.3% than without thermophile bacteria, being hydrolyzed pretreated mud (65 ℃ of thermal treatment).Therefore, thermophile bacteria hydrolyzing sludge fermentation and hydrogen production, can improve and produce the hydrogen rate, and hydrogen purity is high, shortens and produce hydrogen time of lag.
Embodiment 2:
Get marine park organic waste 100ml, the TCOD of waste is 12945mg/L, and SS is 1.74g/L, and VSS is 0.62g/L.Get marine park organic waste 100ml, inoculate Bacillusthermophilic A11 bacterium liquid 10ml of the present invention, regulate pH=7, at 65 ℃, stir speed (S.S.) 140r/min, process 24h, then inoculates Bacillus thermophilic A11 lignin-sludge 10ml, in temperature, be under 35 ℃ of conditions, fermentation and hydrogen production, produce hydrogen after 2h, without the generation of methane, in the biogas produced, the content of hydrogen is greater than 70%, and the maximum hydrogen rate of producing reaches 12.59ml/gCOD.The high-purity hydrogen produced is collected in gas collector, can use after purified.
Claims (4)
1. a bacillus acidocldarius produces the application method in hydrogen in sludge fermentation, comprise sludge pretreatment and ferment for hydrogen production step, it is characterized in that described sludge pretreatment step is inoculation bacillus acidocldarius bacterium liquid in mud, at 50 ~ 70 ℃ of lower pretreating sludge 2 ~ 24 h, obtain bacillus acidocldarius pre-treatment hydrolysis mud; Described bacillus acidocldarius is bacillus acidocldarius Bacillus thermophilic A11 bacterial strain, and its deposit number is CGMCC No.5309; The excess sludge that described mud is sewage work, river/seabed mud or cultivating pool mud; The volume ratio of mud and bacillus acidocldarius bacterium liquid is 100:3～15.
2. application method as claimed in claim 1, it is characterized in that described ferment for hydrogen production step is that bacillus acidocldarius pre-treatment hydrolysis mud is placed in to the ferment for hydrogen production reactor of sealing, to nitrogen blowing 2～15 min in mud, then sealing and fermenting produces hydrogen, and leavening temperature is 25～45 ℃.
3. application method as claimed in claim 1, is characterized in that described bacillus acidocldarius pre-treatment hydrolysis sludge seeding carrys out fermentation and hydrogen production in organic waste; Described organic waste first carries out pre-treatment with bacillus acidocldarius bacterium liquid.
4. application method as claimed in claim 3, is characterized in that the volume ratio of described organic waste and bacillus acidocldarius bacterium liquid is 100:3～15.
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| CN103058478B (en) * | 2013-01-08 | 2014-08-13 | 同济大学 | Method for improving anaerobic digestion performance of sludge and synchronously enlarging cultivation by functional bacteria |
| CN105219667B (en) * | 2014-06-30 | 2018-09-21 | 北京化工大学 | Bacterial strain and hydrogen production process for wood-sugar fermentation hydrogen manufacturing |
| CN106480104B (en) * | 2016-12-07 | 2019-09-06 | 中国科学院生态环境研究中心 | A kind of preprocess method of ferment antibiotics bacteria residue |
| CN106480103B (en) * | 2016-12-07 | 2019-07-26 | 中国科学院生态环境研究中心 | A kind of detoxification method of ferment antibiotics bacteria residue |
| CN106975441A (en) * | 2017-04-28 | 2017-07-25 | 明光市飞洲新材料有限公司 | A kind of method that utilization biological fermentation process is modified to concave convex rod |
| CN106964312A (en) * | 2017-04-28 | 2017-07-21 | 明光市飞洲新材料有限公司 | The preparation method of BOD concave convex rod adsorbent in a kind of removal waste water |
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