CN102776247A - Method for increasing content of propionic acid in batch fermentation acid-producing liquid - Google Patents
Method for increasing content of propionic acid in batch fermentation acid-producing liquid Download PDFInfo
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- CN102776247A CN102776247A CN2012102498987A CN201210249898A CN102776247A CN 102776247 A CN102776247 A CN 102776247A CN 2012102498987 A CN2012102498987 A CN 2012102498987A CN 201210249898 A CN201210249898 A CN 201210249898A CN 102776247 A CN102776247 A CN 102776247A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 124
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 title claims abstract description 108
- 230000004151 fermentation Effects 0.000 title claims abstract description 97
- 235000019260 propionic acid Nutrition 0.000 title claims abstract description 54
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 title claims abstract description 54
- 238000000034 method Methods 0.000 title claims abstract description 18
- 239000002253 acid Substances 0.000 title claims abstract description 12
- 239000007788 liquid Substances 0.000 title abstract description 5
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- 239000000203 mixture Substances 0.000 claims abstract description 48
- 239000010802 sludge Substances 0.000 claims abstract description 40
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- 230000007062 hydrolysis Effects 0.000 claims abstract description 6
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 6
- 239000006228 supernatant Substances 0.000 claims description 64
- 238000010411 cooking Methods 0.000 claims description 59
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 50
- 238000003756 stirring Methods 0.000 claims description 29
- 239000002054 inoculum Substances 0.000 claims description 26
- 235000011187 glycerol Nutrition 0.000 claims description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 239000008399 tap water Substances 0.000 claims description 18
- 235000020679 tap water Nutrition 0.000 claims description 18
- 208000010392 Bone Fractures Diseases 0.000 claims description 17
- 239000012530 fluid Substances 0.000 claims description 17
- 239000012634 fragment Substances 0.000 claims description 17
- 239000004033 plastic Substances 0.000 claims description 17
- 229920003023 plastic Polymers 0.000 claims description 17
- 238000012258 culturing Methods 0.000 claims description 13
- 241000209094 Oryza Species 0.000 claims description 10
- 235000007164 Oryza sativa Nutrition 0.000 claims description 10
- 235000009566 rice Nutrition 0.000 claims description 10
- 235000013372 meat Nutrition 0.000 claims description 9
- 239000003921 oil Substances 0.000 claims description 9
- 235000019198 oils Nutrition 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 4
- 238000010298 pulverizing process Methods 0.000 claims description 4
- 238000009987 spinning Methods 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 230000002053 acidogenic effect Effects 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims 1
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- 239000000243 solution Substances 0.000 claims 1
- 230000007613 environmental effect Effects 0.000 abstract description 4
- 238000002156 mixing Methods 0.000 abstract description 2
- 239000010806 kitchen waste Substances 0.000 abstract 2
- 244000005700 microbiome Species 0.000 abstract 2
- 108090000790 Enzymes Proteins 0.000 abstract 1
- 102000004190 Enzymes Human genes 0.000 abstract 1
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- 239000001963 growth medium Substances 0.000 description 25
- 150000004666 short chain fatty acids Chemical class 0.000 description 16
- 239000012141 concentrate Substances 0.000 description 14
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- 239000002609 medium Substances 0.000 description 12
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- 241000894006 Bacteria Species 0.000 description 4
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- 230000000052 comparative effect Effects 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 239000011574 phosphorus Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 3
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- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of environmental protection and relates to a method for increasing content of propionic acid in batch fermentation acid-producing liquid. The method includes the steps of firstly, mixing kitchen waste and thickened sludge, adding mixture into an anaerobic hydrolysis fermenter for anaerobic fermentation, centrifugally separating mixture, and taking supernate for standby; secondly, activating and enriching propionibacterium seed broth; thirdly, sterilizing the supernate obtained in the step 1, inoculating the propionibacterium seed broth obtained in the step 2 to the supernate, and fermenting in an anaerobic fermenter to generate propionic acid; and fourthly, centrifuging the fermented mixture to obtain the fermented supernate rich in propionic acid after fermentation. The sludge and the kitchen waste are subjected to primary fermentation under the weak alkaline condition, and microorganism extracellular enzyme hydrolysis of macromolecular organic matters such as microorganism exoenzyme hydrolysate, protein and fat is facilitated. The content of the propionic acid in the fermented supernate is 3.56-9.95g COD per L of total acid in the supernate.
Description
Technical field
The invention belongs to environmental protection technical field, relate to a kind of method that batch fermentation produces propionic acid content in the acid solution that improves.
Background technology
Current, body eutrophication is serious day by day, and the nitrogen in the control sewage and the amount of phosphorus are Critical policies that prevents body eutrophication in the world.Applying biological dephosphorization denitrogenation technology can effectively reduce phosphorus and the content of nitrogen in the sewage; And the voltaile fatty acid in the waste water can be biological dephosphorize denitrification required carbon source is provided; There is report (to see Water Science and Technology; 1992,25,185-194) claim that biological process processing 1mg phosphorus needs 6 ~ 9mg short chain fatty acid (SCFAs).
But solvability COD in the water inlet of the many southern municipal wastewater treatment plants of China (comprising SCFAs, particularly acetate and propionic acid) contains quantity not sufficient, is difficult to satisfy the needs of high-performance bio dephosphorization denitrogenation.For this reason, some investigators have studied in batch reactor the mud (comprising primary sludge and excess sludge) that Sewage Plant is produced and carry out the research that anaerobically fermenting prepares short chain fatty acid and (see Chinese invention patent 200510110451.1; Environmental Science and Technology, 2006,40:2025-2029).
According to another bibliographical information (see Water Research, 2004,38:27-36; Bioresource Technology, 2008,99:4400-4407), keeping under the constant prerequisite of the total COD concentration of sewage, the propionic acid/acetate that increases the sewage water inlet than from, can significantly improve the removal effect of nitrogen and phosphorus.This shows that the propionic acid content that increases sewage more helps the raising of saprobiont dephosphorization and denitrification effect than the concentration that increases acetate.
In research process in the past, have been found that through producing to add glucide or rubbish from cooking in the acid system, can improve mud and produce the propionic acid content (Chinese invention patent 200810035585.5 in the acid solution to sludge fermentation; Environmental Science and Technology, 2009,43 (12): 4373-4380).But the content of propionic acid is lower, only less than 50%.
Summary of the invention
The objective of the invention is to provides a kind of method that batch fermentation produces propionic acid content in the acid solution that improves for the defective that overcomes prior art.
For realizing above-mentioned purpose, the present invention adopts following technical scheme:
A kind of method that improves propionic acid content in the batch fermentation product acid solution comprises following steps:
(1) ferment the in advance mixture (fs fermentation) of thickened sludge and rubbish from cooking;
Rubbish from cooking, thickened sludge are mixed, join anaerobically fermenting in the anaerobic hydrolysis fermentor tank, then with the mixture spinning, it is for use to take out supernatant;
(2) activate also enrichment propionibacterium seed liquor;
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant sterilization that step (1) obtains, in the propionibacterium seed liquor that access step (2) obtains, propionic acid is produced in fermentation in anaerobic fermentation tank;
(4) after the fermentation ends, that fermenting mixture is centrifugal, obtain being rich in the fermented supernatant fluid of propionic acid.
In the described step (1); The water ratio of thickened sludge is 98%-99%, and total suspended solid concentration (TS) is 10-20g/L (VS/TS=0.71 ± 0.08), and (wherein VS is an organic substance to be preferably 15g/L; TS is a total suspended matter matter, and VS/TS is the organic content that characterizes mixture).
Described rubbish from cooking is for after rejecting refuses such as chopsticks, broken bone, plastics, the scraps of paper, fragment, and through pulverizing, cross 10 mesh sieves, obtaining C/N is the concentrated rubbish from cooking of 16-20; It is 80% that the adding tap water makes water ratio; The rubbish from cooking main ingredient is a rice, and meat, oils are auxilliary, greengrocery less (can ignore), and wherein, the dry weight of rice, meat, oils is than being (3 ~ 10): (0.5 ~ 5): (0 ~ 2); Preferred rice, the dry weight of meat, oils is than being 7:2:1.
In the described step (1); Thickened sludge and rubbish from cooking by dry weight than (0.3 ~ 0.08): 1 mixes; The total COD (TCOD) that adds after tap water dilutes is 20-40g COD/L, and preferred thickened sludge is 0.18:1 with rubbish from cooking by the dry weight ratio.
In the described step (1), the temperature of anaerobically fermenting is 10～65 ℃, and stirring velocity is 60-250rpm, with Ca (OH)
2The pH of control fermentation is 5-12, and fermentation time is 2-5 days; Preferred leavening temperature is 25 ℃, and the pH of fermentation is 7, and fermentation time is 4 days.
In the described step (1), the rotating speed of spinning is 2000-4000rpm, and centrifugation time is 4-6min.
In the described step (2), propionibacterium (SICC1.256) is activated the back enrichment and is stored in 10%-30% from the inclined-plane glycerine pipe stores the substratum (culture medium prescription is seen table 1), and preserving environment is-50 to-80 ℃; Be seeded to enrichment medium (culture medium prescription is seen table 2) according to inoculum size 5-15% at every turn, in culturing bottle, charge into N
2, for use after 30 ℃ of enrichment 2-4 of temperature days, preferred enrichment 3 days.
In the described step (3), the supernatant that step (1) is obtained moves sterilization 10～40min under 110～130 ℃, 0.06～0.16MPa; Be cooled to room temperature, join in the anaerobism acidogenic fermentation jar behind the adjusting pH 6-8, in supernatant, insert propionibacterium by 5%～15% inoculum size and stir anaerobically fermenting, the time of this anaerobically fermenting is 3 ~ 5 days, keeps pH=5.5 ~ 8.5 (Ca (OH)
2Adjusting), temperature is 25 ~ 35 ℃.
In the described step (4), centrifugal rotation speed is 500-2000rpm, centrifugation time 2-4min.
In the described step (4), propionic acid content is 3.56 ~ 9.95gCOD/L in the fermented supernatant fluid.The kind of rubbish from cooking has a lot, and also influential to the effect of fermentation, the glucide of rice is more; Be converted into propionic acid more easily; And the composition of protein and fat also helps the metabolism growth of mikrobe, and is according to experimental result, more favourable to producing propionic acid under this ratio
The invention has the beneficial effects as follows:
(1) mud and rubbish from cooking are carried out the fs fermentation under weak basic condition, help larger molecular organicses such as extracellular microbial exoenzyme hydrolysis sugar, protein, fat.
(2) propionic acid content that produces of subordinate phase fermentation be total acid 61.2%-74.6% (in the fermented supernatant fluid propionic acid content be in the supernatant total acid 3.56 ~ 9.95gCOD/L), high more a lot of than the propionic acid content of bibliographical information.
Description of drawings
Fig. 1 is the technological synoptic diagram that pure bacterium is improved mud and rubbish from cooking mixed fermentation liquid propionic acid content in the embodiment of the invention.
Embodiment
Further specify the present invention below in conjunction with embodiment and accompanying drawing.
Rubbish from cooking in the following example is for after rejecting refuses such as chopsticks, broken bone, plastics, the scraps of paper, fragment, and through pulverizing, cross 10 mesh sieves, obtaining C/N is the concentrated rubbish from cooking of 16-20.Wherein the dry weight of rice, meat, oils is than being (3 ~ 10): (0.5 ~ 5): (0 ~ 2); Preferred rice, the dry weight of meat, oils is than being 7:2:1.
Embodiment 1
As shown in Figure 1:
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
The rubbish from cooking of crushing screening of learning from else's experience (is rejected in the present embodiments such as chopsticks, broken bone, plastics, the scraps of paper, fragment rubbish from cooking for after rejecting refuses such as chopsticks, broken bone, plastics, the scraps of paper, fragment; Through pulverizing, cross 10 mesh sieves, obtaining C/N is the concentrated rubbish from cooking of 16-20; It is 80% that the adding tap water makes water ratio; The rubbish from cooking main ingredient is a rice, and meat, oils are auxilliary, rice, and meat, oils optimal dry anharmonic ratio are 7:2:1.) 0.41L; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 2.5 days under room temperature, and stirring velocity is 120rpm and uses Ca (OH)
2Control pH is 8.5; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 20% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 20ml glycerine pipe is seeded to 200ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 3 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) that step (2) obtains, anaerobism stirs fermentation 4 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 2min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 9.95gCOD/L (accounts for total short chain fatty acid 74.6%) with mixture.
Embodiment 2
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.37L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.7L and mixes (mud with rubbish from cooking dry weight than being 0.3:1); Add the 1.43L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 2.5 days under room temperature, and stirring velocity is 250rpm, and uses Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 10% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 10ml glycerine pipe is seeded to 100ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 4 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes; Be cooled to room temperature; Inject the 2.5L anaerobic fermentation tank, with Ca (OH) 2 pH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) of step (2) acquisition; Anaerobism stirs fermentation 4 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 2min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 6.10gCOD/L (accounts for total short chain fatty acid 68.2%) with mixture.
Embodiment 3
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.45L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.22L and mixes (mud with rubbish from cooking dry weight than being 0.08:1); Add the 1.83L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 2.5 days under room temperature, and stirring velocity is 60rpm, and uses Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 15% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 30ml glycerine pipe is seeded to 200ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 15%, in culturing bottle, charges into N
2, after 2 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes; Be cooled to room temperature; Inject the 2.5L anaerobic fermentation tank, with Ca (OH) 2 pH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) of step (2) acquisition; Anaerobism stirs fermentation 4 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 2min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 8.70gCOD/L (accounts for total short chain fatty acid 63.9%) with mixture.
Embodiment 4
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 2.5 days under room temperature, and stirring velocity is 120rpm, and uses Ca (OH)
2Control pH is 5; Take out fermenting mixture at the centrifugal 3min of 2000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 30% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 20ml glycerine pipe is seeded to 200ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 3 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes; Be cooled to room temperature; Inject the 2.5L anaerobic fermentation tank, with Ca (OH) 2 pH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) of step (2) acquisition; Anaerobism stirs fermentation 4 days, keeps 30 ℃ of pH=7 (Ca (OH) 2 regulates), temperature.
(4) after the fermentation ends, at the centrifugal 2min of 500rmp, the propionic acid amount in the supernatant of obtaining is 3.56gCOD/L (accounts for total short chain fatty acid 61.2%) with mixture.
Embodiment 5
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 2.5 days under room temperature, and stirring velocity is 120rpm, and uses Ca (OH)
2Control pH is 12; Take out fermenting mixture at the centrifugal 6min of 4000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 20% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 20ml glycerine pipe is seeded to 200ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 3 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) that step (2) obtains, anaerobism stirs fermentation 5 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 4min of 1500rmp, the propionic acid amount in the supernatant of obtaining is 7.50gCOD/L (accounts for total short chain fatty acid 68.9%) with mixture.
Embodiment 6
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 1.5 days under room temperature, and stirring velocity is 120rpm, and uses Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 20% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 30ml glycerine pipe is seeded to 300ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 3 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 40min that under 110 ℃ of high-pressure sterilizing pots, 0.06MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 15%) that step (2) obtains, anaerobism stirs fermentation 4 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 3min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 5.70gCOD/L (accounts for total short chain fatty acid 69.2%) with mixture.
Embodiment 7
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 5 days under room temperature, and stirring velocity is 120rpm, and uses Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 20% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 10ml glycerine pipe is seeded to 200ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 5%, in culturing bottle, charges into N
2, after 4 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 10min that under 130 ℃ of high-pressure sterilizing pots, 0.116MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) that step (2) obtains, anaerobism stirs fermentation 4 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 3min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 7.32gCOD/L (accounts for total short chain fatty acid 64.2%) with mixture.
Embodiment 8
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
In 10 ℃ of fermentations of temperature 2.5 days, stirring velocity was 120rpm, and used Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 10% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 30ml glycerine pipe is seeded to 200ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 15%, in culturing bottle, charges into N
2, after 2 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) that step (2) obtains, anaerobism stirs fermentation 4 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 3min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 5.48gCOD/L (accounts for total short chain fatty acid 71.2%) with mixture.
Embodiment 9
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
In 65 ℃ of fermentations of temperature 2 days, stirring velocity was 120rpm, and used Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 6min of 4000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 20% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 20ml glycerine pipe is seeded to 200ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 3 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 7, and insert the propionibacterium seed liquor (inoculum size 10%) that step (2) obtains, anaerobism stirs fermentation 4 days, keeps pH=7 (Ca (OH)
2Adjusting), temperature is 30 ℃.
(4) after the fermentation ends, at the centrifugal 3min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 9.45gCOD/L (accounts for total short chain fatty acid 73.3%) with mixture.
Embodiment 10
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 2.5 days under room temperature, and stirring velocity is 120rpm, and uses Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 20% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 30ml glycerine pipe is seeded to 300ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 3 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 5.5, and insert the propionibacterium seed liquor (inoculum size 15%) that step (2) obtains, anaerobism stirs fermentation 3 days, keeps pH=5.5 (Ca (OH)
2Adjusting), temperature is 25 ℃.
(4) after the fermentation ends, at the centrifugal 3min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 10.12gCOD/L (accounts for total short chain fatty acid 72.1%) with mixture.
Embodiment 11
(1) mixture (fs fermentation) of preparatory fermented sludge and rubbish from cooking;
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.41L of crushing screening learns from else's experience; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.46L and mixes (mud with rubbish from cooking dry weight than being 0.18:1); Add the 1.63L tap water, be injected into 3L anaerobic fermentation tank (TCOD is 25g COD/L);
(25 ℃) fermentation is 1.5 days under room temperature, and stirring velocity is 120rpm, and uses Ca (OH)
2Control pH is 8; Take out fermenting mixture at the centrifugal 5min of 3000rpm, the 2L centrifuged supernatant of acquisition.
(2) activation and enrichment propionibacterium;
Propionibacterium (SICC1.256) activated the back enrichment and be stored in 20% glycerine pipe store the substratum (culture medium prescription is seen table 1) from the inclined-plane, preserving environment is-80 ℃; Before using 30ml glycerine pipe is seeded to 300ml enrichment medium (culture medium prescription is seen table 2) according to inoculum size 10%, in culturing bottle, charges into N
2, after 3 days, obtain the propionibacterium seed liquor in 30 ℃ of enrichments of temperature.
(3) utilize propionibacterium seed liquor fermented supernatant fluid to produce propionic acid (subordinate phase fermentation);
With the supernatant that arrives of step (1) 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes, be cooled to room temperature, inject the 2.5L anaerobic fermentation tank, with Ca (OH)
2PH is transferred to 8.5, and insert the propionibacterium seed liquor (inoculum size 15%) that step (2) obtains, anaerobism stirs fermentation 3 days, keeps pH=8.5 (Ca (OH)
2Adjusting), temperature is 35 ℃.
(4) after the fermentation ends, at the centrifugal 3min of 1000rmp, the propionic acid amount in the supernatant of obtaining is 6.32gCOD/L (accounts for total short chain fatty acid 68.2%) with mixture.
Comparative Examples 1
Do not inoculate the situation of propionibacterium:
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.25L of crushing screening learns from else's experience; Add the 0.75L tap water; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 1.5L and mixes (mud with rubbish from cooking dry weight than being 0.9:1), be injected into 3L anaerobic fermentation tank (TCOD is 26.90g COD/L), (25 ℃) fermented 8 days under room temperature; Stirring velocity is 120rpm, and uses Ca (OH)
2Control pH is 8.
After the fermentation ends, mixture is centrifugal, and the propionic acid amount in the supernatant of obtaining is 4.73gCOD/L (accounts for total short chain fatty acid 49.1%)
Only with the rubbish from cooking fermentation, mud drops into considerably less the fermentation of 2 fs of Comparative Examples, only does the inoculation of hydrolysis bacterium.
Rubbish from cooking (rejecting chopsticks, broken bone, plastics, the scraps of paper, the fragment etc.) 0.57L of crushing screening learns from else's experience; Add the 1.43L tap water; Concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84) with 0.2L and mixes (dry weight is than being 0.01:1); Be injected into 3L anaerobic fermentation tank (TCOD is 39.08g COD/L), and use Ca (OH)
2Control pH is 8, and (25 ℃) fermentation was taken out fermenting mixture at the centrifugal 4min of 2000rpm after 2.5 days under the room temperature; The 2L centrifuged supernatant that the obtains 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes; Be cooled to room temperature, pH transfers to 7, inserts 200ml enrichment bacterium liquid.Keep 30 ℃ of reaction system pH=7, temperature, anaerobism stirs fermentation 5 days.
After the fermentation ends, mixture is centrifugal, and the propionic acid amount in the supernatant of obtaining is 1.15gCOD/L (accounts for total short chain fatty acid 62.7%).
Sludge fermentation is only used in the fermentation of 3 fs of Comparative Examples, does not relate to rubbish from cooking
Only get 2.5L and concentrate excess sludge (TSS 20g/L, VSS 14g/L, water ratio are 98%, pH=6.84), use Ca (OH)
2Control pH is 8 mixing; (25 ℃) fermentation is after 2.5 days under the room temperature; Take out fermenting mixture at the centrifugal 5min of 2000rpm, the 2L centrifuged supernatant of the acquisition 20min that under 121 ℃ of high-pressure sterilizing pots, 0.110MPa, sterilizes is cooled to room temperature; PH transfers to 7, and 200ml enrichment bacterium liquid is injected 2.5L anaerobism acidogenic fermentation jar simultaneously.Keep 30 ℃ of reaction system pH=7, temperature, anaerobism stirs fermentation 5 days.
After the fermentation ends, mixture is centrifugal, and the propionic acid amount in the supernatant of obtaining is 0.6gCOD/L (accounts for total short chain fatty acid 20.1%).
Embodiment 1-10 compares with the prior art in the Comparative Examples 1 can improve propionic acid content greatly.
Table 1 glycerine stores culture medium prescription
Table 2 enrichment culture based formulas
The above-mentioned description to embodiment is can understand and use the present invention for ease of the those of ordinary skill of this technical field.The personnel of skilled 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 are according to announcement of the present invention, and not breaking away from the improvement that category of the present invention makes and revise all should be within protection scope of the present invention.
Claims (10)
1. one kind is improved the method that batch fermentation produces propionic acid content in the acid solution, it is characterized in that: comprise following steps:
(1) rubbish from cooking, thickened sludge are mixed, join anaerobically fermenting in the anaerobic hydrolysis fermentor tank, then with the mixture spinning, it is for use to take out supernatant;
(2) activate also enrichment propionibacterium seed liquor;
(3) the supernatant sterilization that step (1) is obtained, in the propionibacterium seed liquor that access step (2) obtains, propionic acid is produced in fermentation in anaerobic fermentation tank;
(4) after the fermentation ends, that fermenting mixture is centrifugal, obtain being rich in the fermented supernatant fluid of propionic acid.
2. method according to claim 1 is characterized in that: in the described step (1), the water ratio of thickened sludge is 98%-99%, and total suspended solid concentration is 10-20g/L, is preferably 15g/L.
3. method according to claim 1 is characterized in that: described rubbish from cooking is for after rejecting chopsticks, broken bone, plastics, the scraps of paper, fragment refuse, and through pulverizing, cross 10 mesh sieves, obtaining C/N is the concentrated rubbish from cooking of 16-20; It is 80% that the adding tap water makes water ratio; Wherein, the dry weight of rice, meat, oils is than being (3 ~ 10): (0.5 ~ 5): (0 ~ 2); Preferred rice, the dry weight of meat, oils is than being 7:2:1.
4. method according to claim 1; It is characterized in that: in the described step (1); Thickened sludge and rubbish from cooking by dry weight than (0.3 ~ 0.08): 1 mixes; Preferred thickened sludge is 0.18:1 with rubbish from cooking by the dry weight ratio, and the total COD after the dilution of adding tap water is 20-40g COD/L.
5. method according to claim 1 is characterized in that: in the described step (1), the temperature of anaerobically fermenting is 10～65 ℃, and stirring velocity is 60-250rpm, with Ca (OH)
2The pH of control fermentation is 5-12, and fermentation time is 2-5 days; Preferred leavening temperature is 25 ℃, and the pH of fermentation is 7, and fermentation time is 4 days.
6. method according to claim 1 is characterized in that: in the described step (1), the rotating speed of spinning is 2000-4000rpm, and centrifugation time is 4-6min.
7. method according to claim 1 is characterized in that: in the described step (2), propionibacterium is activated the back enrichment and is stored in 10%-30% from the inclined-plane glycerine pipe stores the substratum, and preserving environment is-50 to-80 ℃; Be seeded to enrichment medium according to inoculum size 5-15% at every turn, in culturing bottle, charge into N
2, for use after 30 ℃ of enrichment 2-4 of temperature days, preferred enrichment 3 days.
8. method according to claim 1 is characterized in that: in the described step (3), the supernatant that step (1) is obtained moves sterilization 10～40min under 110～130 ℃, 0.06～0.16MPa; Be cooled to room temperature, join in the anaerobism acidogenic fermentation jar behind the adjusting pH 6-8, in supernatant, insert propionibacterium by 5%～15% inoculum size and stir anaerobically fermenting, the time of this anaerobically fermenting is 3 ~ 5 days, Ca (OH)
2Keep 25 ~ 35 ℃ of pH=5.5 ~ 8.5, temperature.
9. method according to claim 1 is characterized in that: in the described step (4), centrifugal rotation speed is 500-2000rpm, centrifugation time 2-4min.
10. method according to claim 1 is characterized in that: in the described step (4), propionic acid content is 3.56 ~ 9.95gCOD/L in the fermented supernatant fluid.
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| CN103333925A (en) * | 2013-07-05 | 2013-10-02 | 同济大学 | Rapid production method of fermented acid-producing liquid with high propionic acid content |
| CN103466802A (en) * | 2013-09-03 | 2013-12-25 | 同济大学 | Method for cutting down adverse effects of nano material on sewage biological denitrification and phosphorus removal system |
| CN108060181A (en) * | 2017-12-14 | 2018-05-22 | 天津城建大学 | The method for improving excess sludge and protide rubbish from cooking mixed fermentation production acid |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103333925A (en) * | 2013-07-05 | 2013-10-02 | 同济大学 | Rapid production method of fermented acid-producing liquid with high propionic acid content |
| CN103333925B (en) * | 2013-07-05 | 2015-08-19 | 同济大学 | The method of a kind of quick production high propionic acid content fermentation and acid liquid |
| CN103466802A (en) * | 2013-09-03 | 2013-12-25 | 同济大学 | Method for cutting down adverse effects of nano material on sewage biological denitrification and phosphorus removal system |
| CN103466802B (en) * | 2013-09-03 | 2015-04-15 | 同济大学 | Method for cutting down adverse effects of nano material on sewage biological denitrification and phosphorus removal system |
| CN108060181A (en) * | 2017-12-14 | 2018-05-22 | 天津城建大学 | The method for improving excess sludge and protide rubbish from cooking mixed fermentation production acid |
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