CN101555314A - Method for synthesizing polyhydroxyalkanoates by excess sludge fermentation broth as carbon source - Google Patents
Method for synthesizing polyhydroxyalkanoates by excess sludge fermentation broth as carbon source Download PDFInfo
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Abstract
The invention belongs to the technical field of environment protecting, in particular to a method which uses excess sludge fermentation broth of sewage treatment works as carbon source to synthesize Polyhydroxyalkanoates (PHA) by active sludge in room temperature and aerobic environment. The method firstly hydrolyzes excess sludge generated by sewage treatment works in anaerobic condition to obtain excess sludge fermentation broth; the excess sludge fermentation broth is then added into a reactor comprising active sludge, thereby obtaining high-efficiency accumulation of PHA in the active sludge by Aerobic Feeding and Discharge (AFD) technique. The invention provides a method for effectively synthesizing PHA by recycling excess sludge of sewage treatment works. Therefore, the method has fine environmental and economical benefits.
Description
Technical field
The invention belongs to environmental protection technical field; being specifically related to a kind of is carbon source with sewage treatment plant residual sludge fermentation liquid; utilize active sludge at aerobic repeatedly charging-drainage technology (Aerobic Feeding and Discharge; AFD) synthesizing polyhydroxyalkanoateby (Polyhydroxyalkanoates, method PHA) under the condition.
Background technology
Day by day under the serious situation, PHA is subjected to paying attention to widely as a kind of substitute of traditional chemical synthetic plastics just day by day in the environmental problem of being brought by the non-degradable plastics.This is not only because its mechanical property is similar to conventional plastic, and it can be degraded by biology fully.PHA is energy and the carbon source class stored substance in many microorganism cellss.The PHA of commercial production at present utilizes that to carry out pure-blood ferment through the microorganism of screening or genetic engineering bacterium synthetic,, to compare with traditional petrochemical industry macromolecular material, the production cost of PHA is higher, and this has hindered the business-like process of PHA greatly.As far back as the seventies in 20th century, it has been found that active sludge can accumulate PHA (for example, document Environmental Science ﹠amp; Technology, 1974,8,576-579).Adopt the synthetic PHA of active sludge to need not under aseptic condition, to carry out, thereby production cost is expected to be reduced.
When synthesizing PHA with microorganism, the shared expense of carbon source is another important factor that influences the PHA production cost.In the research in the past, being used for PHA synthetic carbon source mainly is pure substances such as acetate, propionic acid, glucose.For reducing the carbon source cost, just need to seek the more cheap carbon source that to synthesize PHA.Along with the raising of wastewater treatment rate in the worldwide, the excess sludge of annual enormous amount is produced continuously.There are some researches show excess sludge pass through anaerobically fermenting can produce a large amount of short chain fatty acids (Short Chain Fatty Acids, SCFAs), material (for example, document EnvironmentalScience ﹠amp such as carbohydrate and protein; Technology, 2006,40,2025-2029), its Short-Chain Fatty Acids is easily to be utilized and synthesize the material of PHA by microorganism, therefore, the fermented liquid of excess sludge is studied, is developed and have positive effect as the method for PHA synthetic carbon source.
Summary of the invention
The object of the present invention is to provide a kind of fermented liquid of giving birth to the sewage treatment plant residual sludge anaerobic fermentation to produce is carbon source, utilizes the method for active sludge synthesizing poly (hydroxyalkanoate) (PHA).
For achieving the above object, solution of the present invention is:
At the characteristics of fermented liquid component, (Aerobic Feeding and Discharge AFD) carries out the synthetic of PHA, and has obtained higher output (PHA accounts for dry cell weight 72.9%) to adopt aerobic repeatedly charging-drainage technology.So-called AFD technology is divided into fermented liquid several times exactly and adds in the reactor, and the SCFAs in the fermented liquid of at every turn waiting to be added is exhausted, stop aeration and staticly settle, discharge the supernatant liquor with add of fermented liquid equal volume, then add isopyknic fermented liquid again, restart aeration.AFD technology provides the outer carbon source that alternately occurs sufficient and shortage environment for active sludge, and this environment helps the synthetic and accumulation of PHA.Simultaneously, discharge supernatant liquor in the gap of twice afterfermentation liquid and can guarantee SCFAs concentration higher in the charging post-reactor again, higher organic loading helps the synthetic (document: Journal of Chemical Technology﹠amp of PHA; Biotechnology, 2005,80,1306-1318).Therefore, AFD technology provides the environment that is suitable for synthetic PHA for active sludge, thereby has obtained higher output.
The a kind of of the present invention's proposition is carbon source with the short chain fatty acid in the sewage treatment plant residual sludge fermentation liquid, utilize active sludge at aerobic repeatedly charging-drainage technology (Aerobic Feeding and Discharge, AFD) method of synthetic PHA under the condition, concrete steps are as follows:
(1) get the excess sludge that sewage work produces and put into fermentor tank, regulate pH to 8-11, at room temperature stir 6-10d (my god), mixture is carried out mud-water separation, the liquid of collecting is fermented liquid.
(2) active sludge is placed reactor, to wherein adding the fermented liquid that obtains in (1), aeration under aerobic condition, SCFAs in the fermented liquid of waiting to be added is exhausted, stop aeration and staticly settle, discharge with abovementioned steps in institute add the supernatant liquor of fermented liquid equal volume, then add also aeration once more of isopyknic fermented liquid.Repeat the said process several times, treat that the SCFAs in the last fermented liquid that adds is exhausted, stop aeration.
(3) accumulated the active sludge of PHA by centrifugal collection, mix concussion with chloroform after lyophilize, got its supernatant liquor evaporation concentration in Rotary Evaporators, then, added cold ethanol under agitation condition, the flocks of gained white is synthetic PHA.
Wherein, the method for excess sludge anaerobically fermenting production fermented liquid adopts reported method (for example, document Environmental Science ﹠amp in the document in (1); Technology, 2006,40,2025-2029); Reported method in the PHA extracting method employing document (3) (for example: document: Macromolecule, 1986,19:2860-2864).
In above-mentioned PHA building-up process, the too high meeting of carbon source concentration causes inhibition to active sludge, crosses the low then corresponding reduction of semi-invariant of PHA, and therefore, the ratio of fermented liquid and active sludge was controlled at 0.5: 1～10: 1 usually.Simultaneously, suitable dissolved oxygen concentration also is to promote SCFAs to be absorbed by active sludge and be converted into one of important factor of PHA, is controlled at 0.5～8mg/L usually.In addition, too short aeration time can not guarantee the abundant absorption of SCFAs, and long aeration time can cause the decomposition of cumulative PHA in the active sludge, and therefore, aeration time is controlled at 0.5～10h usually.At last, suitable fermented liquid adds the utilising efficiency that number of times can improve carbon source in the fermented liquid on the one hand, can ensure that on the other hand active sludge obtains competent carbon source, is controlled at usually 2～10 times.
Wherein, good conditions of the present invention is: the volume ratio of fermented liquid and active sludge is 1: 1～5: 1; Add fermented liquid aeration time 1～5h at every turn; Dissolved oxygen concentration is controlled at 4～7mg/L; It is 3～6 times that fermented liquid adds number of times.
Owing to adopted such scheme, the invention has the beneficial effects as follows:
(1) has good benefits in environment: at first, when producing fermented liquid, can realize mud decrement and stabilization; Secondly, do carbon source, can reduce the consumption of Nonrenewable energy resources such as acetate, glucose with fermented liquid; At last, the synthetic PHA of institute is a completely biodegradable material, is used for substituting the environmental pollution that traditional petrochemical industry plastics can solve conventional plastic to a certain extent and caused;
(2) has good economic benefit: on the one hand, adopt AFD technology can obtain higher PHA output, thereby can reduce the production cost of PHA; On the other hand, activated sludge process need not strict red-tape operati condition, can cost saving.
Embodiment
The present invention is further illustrated below in conjunction with embodiment.
Embodiment 1:
(1) excess sludge anaerobism alkaline fermentation is produced SCFAs
The method of excess sludge anaerobism alkaline fermentation production SCFAs with reference to pertinent literature (for example, document: EnvironmentalScience and Technology, 2006,40:2025-2029), concrete steps are: in the working volume that synthetic glass is made is 5 liters reactor, (its water ratio is 99.4% to the excess sludge of adding municipal sewage plant, pH=6.72) as the substrate of fermentation and acid, temperature of reaction is 20 ± 1 ℃, adjusting pH is 10, under anaerobic stirs 7d, and it is centrifugal to get mud mixture, collect clear liquid and be fermented liquid, be used as the carbon source of following PHA in synthetic.Contain SCFAs 2606mgCOD/L in the fermented liquid, protein 882mg/L, carbohydrate 201mg/L.
(2) the synthetic PHA of active sludge
The 0.5L active sludge is placed reactor, simultaneously to wherein adding fermented liquid, aeration under aerobic condition, the control dissolved oxygen concentration is 8mg/L, institute adds fermentating liquid volume and the active sludge volume ratio is 0.5: 1, add fermented liquid altogether 5 times, treat at every turn add after SCFAs in the fermented liquid is exhausted, stop aeration and staticly settle, discharge with abovementioned steps in add the supernatant liquor of fermented liquid equal volume, and add isopyknic fermented liquid, distinguish aeration 0.5 after adding fermented liquid for 5 times, 0.5,1,1,2h, then stop aeration, centrifugal collection active sludge records in every gram mud and contains PHA317.4mg.
(3) mensuration of PHA in the active sludge
The PHA Determination on content with reference to pertinent literature (for example in the active sludge, document: Applied Microbiology andBiotechnology, 1978,6:29-37), concrete steps are: be dried to mud sample Powdered in freeze drier, take by weighing dried mud powder in the cracking bottle, add chloroform, sulfuric acid-methyl alcohol and phenylformic acid-methanol solution successively, in 102 ℃ of following cracking a few hours, after cooling, adding distil water is centrifugal, gets organic phase and carries out gas chromatographic analysis.
(4) extraction of PHA in the active sludge
The extracting method of PHA with reference to pertinent literature (for example in the active sludge, document Macromolecule, 1986,19:2860-2864), concrete steps are: the cryodesiccated mud sample of learning from else's experience, mix concussion 24h (30 ℃ of shaking tables with the chloroform of 10 times of volumes, 200 commentaries on classics/min), above-mentioned mixed solution is centrifugal, promptly get the supernatant liquor that contains PHA, supernatant liquor after the evaporation concentration, is added 5 times of cold ethanol of volume (0～4 ℃) and place 30min in 0 ℃ of refrigerator in Rotary Evaporators under agitation condition.Treat that white flocks occurs, filtering drying promptly gets the synthetic PHA of institute.
Embodiment 2:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 1: 1, the control dissolved oxygen concentration is 0.5mg/L, distinguish aeration 3,4,5,6,7h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA186.3mg.
Embodiment 3:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 10: 1, the control dissolved oxygen concentration is 6mg/L, it is 3 times that fermented liquid adds number of times, distinguish aeration 6,7,8h after adding fermented liquid at every turn, all the other are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA292.5mg.
Embodiment 4:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 8: 1, the control dissolved oxygen concentration is 8mg/L, it is 6 times that fermented liquid adds number of times, distinguish aeration 5,6,7,8,9,10h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA354.6mg.
Embodiment 5:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 1: 1, the control dissolved oxygen concentration is 6mg/L, it is 2 times that fermented liquid adds number of times, distinguish aeration 1,2h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA374.9mg.
Embodiment 6:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 5: 1, the control dissolved oxygen concentration is 7mg/L, it is 3 times that fermented liquid adds number of times, distinguish aeration 4,5,6h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA408.6mg.
Embodiment 7:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 2: 1, the control dissolved oxygen concentration is 4mg/L, it is 4 times that fermented liquid adds number of times, distinguish aeration 2,3,4,5h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA423.8mg.
Embodiment 8:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 0.5: 1, the control dissolved oxygen concentration is 5mg/L, it is 10 times that fermented liquid adds number of times, distinguish aeration 1,2,3,4,5,6,7,8,9,10h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA485.2mg.
Embodiment 9:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 3: 1, the control dissolved oxygen concentration is 6mg/L, it is 3 times that fermented liquid adds number of times, distinguish aeration 2.5,3.5,5h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA548.3mg.
Embodiment 10:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 1: 1, the control dissolved oxygen concentration is 6mg/L, it is 3 times that fermented liquid adds number of times, distinguish aeration 1,2,3 after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA647.9mg.
Embodiment 11:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 1: 1, the control dissolved oxygen concentration is 6mg/L, it is 4 times that fermented liquid adds number of times, distinguish aeration 1,2,3,4h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA729.1mg.
Embodiment 12:
The mensuration of PHA and extracting method are seen embodiment 1 in the acquisition of excess sludge fermentation broth and the mud.
The synthetic method of PHA is as follows: change the volume ratio of the fermented liquid that adds and active sludge be 1: 1, the control dissolved oxygen concentration is 6mg/L, distinguish aeration 1,2,3,4,5h after adding fermented liquid at every turn, all the other conditions are with embodiment 1, collect active sludge when reaction finishes, record in every gram mud and contain PHA732.4mg.The weight-average molecular weight that the 150C type gel permeation chromatography of employing Waters company records gained PHA is 8.5 * 10
5Da, the fusing point that the Q100 type difference formula scanning calorimeter of employing TA company records gained PHA is 101.4 ℃, glass transition temp is 2.68 ℃.
Above-mentioned description to embodiment is can understand and apply the invention for the ease of those skilled in the art.The person skilled in the art obviously can easily make various modifications to these embodiment, and needn't pass through performing creative labour being applied in the General Principle of this explanation among other embodiment.Therefore, the invention is not restricted to the embodiment here, those skilled in the art should be within protection scope of the present invention for improvement and modification that the present invention makes according to announcement of the present invention.
Claims (7)
1. the method for a synthesizing polyhydroxyalkanoateby, it is characterized in that: with sewage treatment plant residual sludge fermentation liquid is carbon source, utilizes active sludge synthesizing polyhydroxyalkanoateby under aerobic repeatedly charging-drainage technology condition.
2. method according to claim 1 is characterized in that:
(1) excess sludge of getting sewage work's generation is put into fermentor tank, regulates pH to 8～11, at room temperature stirs 6～10d, and mixture is carried out mud-water separation, and the liquid of collecting is fermented liquid;
(2) active sludge is placed reactor, the fermented liquid that in wherein adding step (1), obtains, aeration under aerobic condition, short chain fatty acid in the fermented liquid of waiting to be added is exhausted, stop aeration and staticly settle, discharge the supernatant liquor with add of fermented liquid equal volume, then add also aeration once more of isopyknic fermented liquid; Repeat the said process several times, treat that the short chain fatty acid in the last fermented liquid that adds is exhausted, stop aeration;
(3) accumulated the active sludge of polyhydroxyalkanoate by centrifugal collection, after lyophilize, mix concussion, get its supernatant liquor evaporation concentration in Rotary Evaporators, then with chloroform, add cold ethanol under agitation condition, the flocks of gained white is the synthetic polyhydroxyalkanoate.
3. method according to claim 2 is characterized in that: the fermented liquid that is added in the step (2) and the volume ratio of active sludge are 0.5: 1～10: 1.
4. method according to claim 2 is characterized in that: the aeration time in the step (2) after each adding fermented liquid is exhausted with short chain fatty acid and is as the criterion aeration 0.5～10h.
5. method according to claim 2 is characterized in that: in the aeration process, dissolved oxygen concentration is 0.5～8mg/L in the reactor in the step (2).
6. method according to claim 2 is characterized in that: the number of times that adds of fermented liquid is 2-10 time in the step (2).
7. method according to claim 2 is characterized in that: the volume ratio of fermented liquid and active sludge is 1: 1～5: 1; Add fermented liquid aeration time 1～5h at every turn; Dissolved oxygen concentration is controlled at 4～7mg/L; It is 3～6 times that fermented liquid adds number of times.
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| CN101735440B (en) * | 2009-12-24 | 2011-12-14 | 哈尔滨工业大学 | Method for synthesizing polyhydroxy alkanoates by excess sludge in water treatment |
| US20120006758A1 (en) * | 2010-07-09 | 2012-01-12 | National Chi Nan University | Method of recovering polyhydroxyalkanoates (phas) from municipal waste sludge |
| CN102505025A (en) * | 2011-12-02 | 2012-06-20 | 厦门大学 | Method for synthesizing polyhydroxyalkanoate by using residual sludge broth as substrate |
| CN103290073A (en) * | 2013-05-20 | 2013-09-11 | 同济大学 | Method for synthesizing polyhydroxyalkanoate with high hydroxyvalerate content |
| CN107207710A (en) * | 2015-02-27 | 2017-09-26 | 阿弗于伦公司 | For by can the precursor method for preparing PHA that obtains of fermentation of biomass by anaerobic fermentation |
| CN108866116A (en) * | 2018-07-12 | 2018-11-23 | 同济大学 | Extend the method that medium chain fatty acid is prepared as raw material using organic waste using carbochain |
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| CN101255227A (en) * | 2008-04-14 | 2008-09-03 | 邵胜学 | Method for synthesizing polyhydroxyalkanoates by employing oil-containing sludge |
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| CN101735440B (en) * | 2009-12-24 | 2011-12-14 | 哈尔滨工业大学 | Method for synthesizing polyhydroxy alkanoates by excess sludge in water treatment |
| US20120006758A1 (en) * | 2010-07-09 | 2012-01-12 | National Chi Nan University | Method of recovering polyhydroxyalkanoates (phas) from municipal waste sludge |
| US8641907B2 (en) * | 2010-07-09 | 2014-02-04 | National Chi Nan University | Method of recovering polyhydroxyalkanoates (PHAs) from municipal waste sludge |
| CN102505025B (en) * | 2011-12-02 | 2014-09-24 | 厦门大学 | Method for synthesizing polyhydroxyalkanoate by using residual sludge broth as substrate |
| CN102505025A (en) * | 2011-12-02 | 2012-06-20 | 厦门大学 | Method for synthesizing polyhydroxyalkanoate by using residual sludge broth as substrate |
| CN103290073B (en) * | 2013-05-20 | 2014-12-10 | 同济大学 | Method for synthesizing polyhydroxyalkanoate with high hydroxyvalerate content |
| CN103290073A (en) * | 2013-05-20 | 2013-09-11 | 同济大学 | Method for synthesizing polyhydroxyalkanoate with high hydroxyvalerate content |
| US10465214B2 (en) | 2014-11-20 | 2019-11-05 | Full Cycle Bioplastics Llc | Producing resins from organic waste products |
| US11377672B2 (en) | 2014-11-20 | 2022-07-05 | Full Cycle Bioplastics Llc | Producing resins from organic waste products |
| CN107207710A (en) * | 2015-02-27 | 2017-09-26 | 阿弗于伦公司 | For by can the precursor method for preparing PHA that obtains of fermentation of biomass by anaerobic fermentation |
| CN107207710B (en) * | 2015-02-27 | 2019-11-08 | 阿弗于伦公司 | For by by anaerobic fermentation can fermentation of biomass obtain precursor prepare polyhydroxyalkanoates method |
| CN108866116A (en) * | 2018-07-12 | 2018-11-23 | 同济大学 | Extend the method that medium chain fatty acid is prepared as raw material using organic waste using carbochain |
| CN108866116B (en) * | 2018-07-12 | 2022-04-05 | 同济大学 | Method for preparing medium-chain fatty acid by using carbon chain extension and using organic waste as raw material |
| CN112813112A (en) * | 2021-01-07 | 2021-05-18 | 上海碧州环保能源科技有限公司 | Non-methanation process with PHA production as guide |
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