CN101254969B - Method for preparing microbial flocculant by using byproduct bacterial of fermentation industry - Google Patents
Method for preparing microbial flocculant by using byproduct bacterial of fermentation industry Download PDFInfo
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- 230000004151 fermentation Effects 0.000 title claims abstract description 46
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- 238000000034 method Methods 0.000 title claims abstract description 22
- 239000006227 byproduct Substances 0.000 title claims description 18
- 230000001580 bacterial effect Effects 0.000 title description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 67
- 239000007788 liquid Substances 0.000 claims abstract description 45
- 235000011187 glycerol Nutrition 0.000 claims abstract description 30
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 25
- 239000008103 glucose Substances 0.000 claims abstract description 25
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 16
- 238000011218 seed culture Methods 0.000 claims abstract description 12
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 11
- 238000005516 engineering process Methods 0.000 claims abstract description 9
- 239000001963 growth medium Substances 0.000 claims abstract description 9
- 239000007787 solid Substances 0.000 claims abstract description 9
- 239000002609 medium Substances 0.000 claims abstract description 8
- 238000000926 separation method Methods 0.000 claims abstract description 8
- 238000010564 aerobic fermentation Methods 0.000 claims abstract description 6
- 238000005374 membrane filtration Methods 0.000 claims abstract description 5
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 238000002156 mixing Methods 0.000 claims description 14
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 13
- 238000012262 fermentative production Methods 0.000 claims description 9
- 239000002054 inoculum Substances 0.000 claims description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 5
- 239000012670 alkaline solution Substances 0.000 claims description 4
- 238000004062 sedimentation Methods 0.000 claims description 3
- 239000002351 wastewater Substances 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 230000002779 inactivation Effects 0.000 claims description 2
- 238000005507 spraying Methods 0.000 claims description 2
- 238000003860 storage Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 9
- 230000003311 flocculating effect Effects 0.000 abstract description 8
- 239000000126 substance Substances 0.000 abstract description 8
- 241000894006 Bacteria Species 0.000 abstract description 3
- 238000010612 desalination reaction Methods 0.000 abstract description 2
- 238000004821 distillation Methods 0.000 abstract description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 abstract 4
- YPFDHNVEDLHUCE-UHFFFAOYSA-N propane-1,3-diol Chemical compound OCCCO YPFDHNVEDLHUCE-UHFFFAOYSA-N 0.000 abstract 4
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 abstract 2
- 241000588748 Klebsiella Species 0.000 abstract 1
- 238000010364 biochemical engineering Methods 0.000 abstract 1
- 238000005119 centrifugation Methods 0.000 abstract 1
- 238000001953 recrystallisation Methods 0.000 abstract 1
- UXFQFBNBSPQBJW-UHFFFAOYSA-N 2-amino-2-methylpropane-1,3-diol Chemical compound OCC(N)(C)CO UXFQFBNBSPQBJW-UHFFFAOYSA-N 0.000 description 11
- 238000005189 flocculation Methods 0.000 description 11
- 230000016615 flocculation Effects 0.000 description 11
- 239000000047 product Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
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- 238000002360 preparation method Methods 0.000 description 5
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- 239000002551 biofuel Substances 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
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- 230000000694 effects Effects 0.000 description 3
- 239000005995 Aluminium silicate Substances 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 235000012211 aluminium silicate Nutrition 0.000 description 2
- 238000004042 decolorization Methods 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 239000008394 flocculating agent Substances 0.000 description 2
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
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- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
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- 229920002521 macromolecule Polymers 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
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- Separation Of Suspended Particles By Flocculating Agents (AREA)
Abstract
The invention relates to a method for preparing a microbial flocculant by using accessory substance thallus in the fermentation industry, which belongs to the biochemical engineering technology field. The invention adopts the processes that klebsiella bacillus is added into a seed culture medium containing glycerin or glucose, then the seed culture medium is mixed into an initial concentration fermentation medium containing glycerin or glucose, to perform anaerobic fermentation or aerobic fermentation, after the fermentation production is finished, solid-liquid separation is performed to the fermentation liquor through membrane filtration or centrifugation means, the obtained solid part can be directly used as the microbial flocculant, and 1, 3-propylene glycol and 2.3 butyleneene glycol can be separated from the clear liquid and be purified through desalination, distillation, vacuum recrystallization, and other steps to prepare the finished products of 1, 3-propylene glycol and 2.3 butyleneene glycol. The method has the advantages that the thallus is directly sourced from the fermentation industry, the special culture to the flocculating bacteria is not required, so that the production cost of the microbial flocculant is greatly reduced, and meanwhile a way for the value-added utilization of the accessory substance in the fermentation industry is found.
Description
Technical field
The invention belongs to technical field of biochemical industry, particularly a kind of by product thalline that utilizes fermentation industry prepares the method for microbial flocculant.
Background technology
Flocculating Settling Technology is the increase water quality a kind of not only economy but also easy treatment process of processing efficiency, be widely used in Industrial Wastewater Treatment, give in the industry such as water treatment, foodstuff production and fermentation, along with the pay attention to day by day of people's environmental pollution problem and improving constantly of human living standard, water quality has been proposed more and more higher requirement, wide stage is provided for simultaneously the application of Flocculating Settling Technology and development.At present; flocculation agent is divided into inorganic flocculating agent; three kinds of organic floculant and microbial flocculants; microbial flocculant is a kind of macromolecular compound with flocculation activity by microorganisms; can make the solid suspended particle in the liquid; metal ion; somatic cells and colloidal particle etc. condense; sedimentation; by contrast; the microbial flocculant that utilizes biotechnology to obtain has efficiently; safety non-toxic; series of advantages such as non-secondary pollution; protection to human beings'health and environment has very important practical sense; therefore, microbial flocculant has broad application prospects.
The bacterium for producing flocculant of microbe wide material sources, kind is more.So far nearly 80 kinds of (Gan Li, Meng Zhaoping of the microorganism with flocculence of Fa Xianing, the progress of microbial flocculant, water technology, 2006-04:5-9), wherein bacterium, fungi, actinomycetes and algae all have distribution, and the microorganism that these have flocculation activity, paid close attention to by the people is mostly screened from soil, mud, sewage.At present, the preparation of microbial flocculant utilizes sugar or agricultural byproducts as raw materials for production more, and cost is higher, has restricted microbial flocculant suitability for industrialized production and large-scale application.The solution of this bottleneck problem need be started with from two aspects: the one, actively seek cheap starting material, reduce production costs significantly, the 2nd, at the physico-chemical property and the purposes of biofloculation, select rational industrialization extraction process and using method (Ma Fang etc., compound biological flocculant, China Patent No.: 200610010369.6), based on this, the present invention turns to the industrial fermentation industry with sight, utilize the by product thalline of fermentation industry, directly be developed to microbial flocculant, not only greatly reduce the production cost of microbial flocculant, also found an outlet simultaneously for the increment utilization of fermentation industry by product.
1, ammediol and 2.3-butyleneglycol are important chemical material, become the focus that people pay close attention to day by day, and its production method has chemical synthesis and microbe fermentation method, by contrast, microbe fermentation method has mild condition, easy and simple to handle, advantage such as by product is few, environmental pollution is little.Along with Nonrenewable resources reduce and the increasing of world population day by day, green industry with the microbe fermentation method by representative is more and more paid attention to by people, the present invention is a main raw material fermentative production 1 by utilizing with glycerine or glucose at first, and the by product thalline of ammediol and 2.3-butyleneglycol prepares microbial flocculant.
Summary of the invention
The purpose of this invention is to provide the method that a kind of by product thalline that utilizes fermentation industry prepares microbial flocculant, as: utilizing with glycerine or glucose is main raw material fermentative production 1, the by product thalline of ammediol and 2.3-butyleneglycol prepares microbial flocculant, concrete technology is Cray Bai Shi bacillus to be inserted contain in the seed culture medium of glycerine or glucose, cultivate 12h～24h for 30 ℃～37 ℃, again with seed liquor by volume 1～10% inoculum size add and contain glycerine or glucose concn is the initial fermention medium of 30～80g/l, carry out anaerobism or aerobic fermentation under 30～40 ℃ of conditions, feed 0～1.0vvm air during the fermentation, mixing speed 30～350rpm, stream glycerol adding or glucose make in the fermented liquid glycerine or glucose concn be controlled at 10～50g/l.And adopt stream to add 3～4M alkaline solution and control pH value 5.0～8.0.After fermentative production finishes, adopt membrane filtration, means such as centrifugal with the fermented liquid solid-liquid separation, clear liquid passes through conventional steps such as desalination, distillation and rectification under vacuum with 1, ammediol separates, purifies with the 2.3-butyleneglycol, be prepared into product 1, ammediol and 2.3-butyleneglycol, solid (can contain a small amount of fermented liquid) the part thalline that just ferments just can directly use as microbial flocculant.Also can handle or spraying drying by 80～105 ℃ of simple heat inactivations, so that storage and transport are little to its flocculation activity influence.
During application, waste water per ton adds 0～500 gram calcium chloride earlier, adds 0.1～1 kilogram of undried microbial flocculant (or the spray-dired microbial flocculant of 20～200 grams) again, uses sodium hydroxide adjust pH 4.0～8.0 behind the mixing, abundant again mixing, standing sedimentation gets final product.
The invention has the advantages that, the by product thalline that microbial flocculant directly produces from fermentation industry, it has not only found an outlet for the increment utilization of fermentation industry by product, and need not specially the flocculation thalline to be cultivated again, greatly reduces the production cost of microbial flocculant.
Embodiment
The purpose of this invention is to provide the method that a kind of by product thalline that utilizes fermentation industry prepares microbial flocculant, with specific embodiment the present invention further specified again below:
Example 1
(1) bacterial classification: Cray Bai Shi bacillus
(2) fermentation mode: 5L mechanical agitating fermentation tank, reagent glycerine are raw material,
(3) fermentative production product: 1, ammediol, a small amount of 2.3-butyleneglycol of by-product.
A. substratum:
* the preparation of ferrous solution: add FeSO in every premium on currency
4H
2O 5.0g, 37% concentrated hydrochloric acid 4ml.
The composition of * trace element solution
B. fermenting process:
Seed culture: Cray Bai Shi bacillus is inserted in the seed culture medium that contains 30g/L glycerine (250ml triangular flask, liquid amount 50ml) 30 ℃ of culture temperature, shaking speed 140rpm, aerobic cultivation 24h.
Fermentation culture: fermentation culture adopts the 5L fermentor tank, liquid amount 4L, and 37 ℃ of culture temperature are with sodium hydroxide control pH value 6.8.Seed liquor 1% inoculum size access is by volume contained in the initial fermention medium of 30g/L glycerine, flow glycerol adding in the fermenting process, glycerol concentration is about 20g/L in the control fermented liquid.Adopt micro-aerobe fermentation, feed the 0.2vvm air, rotating speed 250rpm.During the 48hr fermentation ends, in the fermented liquid 1, ammediol concentration 81.53g/l, 2.3-butyleneglycol concentration 17.28g/l.
(4) preparation of microbial flocculant and application:
After 48 hours fermentation ends, getting fermented liquid leaves the heart with whizzer 10000 and made its solid-liquid separation in 1 minute, clear liquid partly proceeds 1, the extraction of ammediol product, solid part adds the water with centrifuged supernatant equivalent, mixing is stand-by, in 100 milliliter 0.4% kaolin, adds 1 milliliter of 1% calcium chloride solution earlier, add 2 milliliters of microorganism flocculation liquid again, adjust pH fully behind the mixing, did not leave standstill 6 minutes, originally the rapid conglomeration of Hun Zhuo kaolin suspension liquid precipitates, solid-liquid interface is clear, gets the OD value under the supernatant liquor mensuration 550nm, calculates flocculating rate.The result is as follows:
| The pH value | OD550 | Flocculating rate % | |
| After flocculating before the flocculation | 6.5 6.5 | 2.5416 0.1350 | 94.69 |
Example 2
(1) bacterial classification: Cray Bai Shi bacillus
(2) fermentation mode: 5L mechanical agitating fermentation tank, glucose are raw material.
(3) fermentative production product: 2, the 3-butyleneglycol.
A. substratum:
B. fermenting process:
Seed culture: Cray Bai Shi bacillus is inserted in the seed culture medium that contains 30g/L glucose (250ml triangular flask, liquid amount 50ml) 37 ℃ of culture temperature, shaking speed 180rpm, aerobic cultivation 24h.
Fermentation culture: fermentation culture adopts the 5L mechanical agitating fermentation tank, liquid amount 4L, and 37 ℃ of culture temperature are with sodium hydroxide control pH value 6.0.With seed liquor by volume 10% inoculum size insert and to contain in the initial fermention medium of 150g/L glucose, to add mass concentration be 50% glucose solution to stream in the fermenting process, glucose concn is about 20g/L in the control fermented liquid.Adopt aerobic fermentation, feed the 0.5vvm air, rotating speed 350rpm.During the 72hr fermentation ends, 2.3-butyleneglycol concentration 87.36g/l in the fermented liquid.
(4) preparation of microbial flocculant and application: after 72 hours fermentation ends, fermented liquid filters with metallic membrane and makes its solid-liquid separation, clear liquid partly proceeds 2, the extraction of 3-butyleneglycol product, solid (containing a small amount of fermented liquid) adds and the water of crossing cleaner liquid equivalent, stand-by after 121 ℃ of 20 minutes sterilising treatment behind the mixing, in the blue ink of 30 times of 100 milliliters of dilutions, add 1 milliliter of 1% calcium chloride solution earlier, add 2 milliliters of microorganism flocculation liquid again, fully mixing, leave standstill and promptly produce significantly flocculation phenomenon a moment, blue throw out precipitation after 30 minutes, supernatant fluid color is removed substantially, and the colorimetric estimation percent of decolourization reaches 85.79%.
Example 3
(1) bacterial classification: Cray Bai Shi bacillus
(2) fermentation mode: 40L gas lift fermentor tank, biofuel accessory substance glycerine is raw material.
(3) fermentative production product: 1, ammediol, a small amount of 2.3-butyleneglycol of by-product.
A. substratum: with example 1
B. fermenting process:
Seed culture: Cray Bai Shi bacillus is inserted in the seed culture medium that contains 30g/L biofuel accessory substance glycerine (250ml triangular flask, liquid amount 50ml) 30 ℃ of culture temperature, shaking speed 140rpm, aerobic cultivation 24h.
Fermentation culture: fermentation culture adopts 40L gas lift fermentor tank, liquid amount 30L, and 37 ℃ of culture temperature are with sodium hydroxide control pH value 6.8.Seed liquor 1% inoculum size access is by volume contained in the initial fermention medium of 30g/L biofuel accessory substance glycerine, and stream adds biofuel accessory substance glycerine in the fermenting process, and glycerol concentration is about 20g/L in the control fermented liquid.Adopt micro-aerobe fermentation, feed the 0.6vvm air.During the 48hr fermentation ends, in the fermented liquid 1, ammediol concentration 78.90g/l, 2.3-butyleneglycol concentration 20.35g/l.
(4) preparation of microbial flocculant and application: after 48 hours fermentation ends, getting fermented liquid filters with ceramic membrane and makes its solid-liquid separation, clear liquid partly proceeds 1, the extraction of ammediol product, solid part adds the water with centrifuged supernatant equivalent, mixing is stand-by, be in the waste water of blueness in 100 milliliters of certain cloth dyeing and finishing factories, add 1 milliliter of 1% calcium chloride solution earlier, add 2 milliliters of microorganisms flocculation liquid again, shake up, with sodium hydroxide adjust pH to 7.5, fully behind the mixing, left standstill 15 minutes, get the OD value under the supernatant liquor mensuration 550nm, calculate flocculating rate.The result is as follows:
| The pH value | OD550 | CODCr | Percent of decolourization % | |
| After flocculating before the flocculation | 6.90 7.42 | 0.948 0.065 | 418 118 | 93.14 |
Claims (3)
1. a by product thalline that utilizes fermentation industry prepares the method for microbial flocculant, it is characterized in that, technology is: the access of Cray Bai Shi bacillus is contained in the seed culture medium of glycerine or glucose, cultivate 12h～24h for 30 ℃～37 ℃, again with seed liquor by volume the inoculum size of 1-10% add and to contain glycerine or glucose concn is the initial fermention medium of 30～80g/L, carry out anaerobism or aerobic fermentation under 30～40 ℃ of conditions, feed 0～1.0vvm air during the fermentation, mixing speed 30～350rpm, stream glycerol adding or glucose make in the fermented liquid glycerine or glucose concn be controlled at 10～50g/L; And adopt stream to add 3～4M alkaline solution and control pH value 5.0～8.0; After fermentative production finishes, adopt membrane filtration, centrifugal means with the fermented liquid solid-liquid separation, solid part directly uses as microbial flocculant.
2. a by product thalline that utilizes fermentation industry prepares the method for microbial flocculant, it is characterized in that, technology is: the access of Cray Bai Shi bacillus is contained in the seed culture medium of glycerine or glucose, cultivate 12h～24h for 30 ℃～37 ℃, again with seed liquor by volume the inoculum size of 1-10% add and to contain glycerine or glucose concn is the initial fermention medium of 30～80g/L, carry out anaerobism or aerobic fermentation under 30～40 ℃ of conditions, feed 0～1.0vvm air during the fermentation, mixing speed 30～350rpm, stream glycerol adding or glucose make in the fermented liquid glycerine or glucose concn be controlled at 10～50g/L; And adopt stream to add 3～4M alkaline solution and control pH value 5.0～8.0; After fermentative production finishes, adopt membrane filtration, centrifugal means with the fermented liquid solid-liquid separation; Solid part is handled or spraying drying at 80～105 ℃ of heat inactivations, so that storage and transport.
3. a by product thalline that utilizes fermentation industry prepares the method for microbial flocculant, its spy just is, technology is: the access of Cray Bai Shi bacillus is contained in the seed culture medium of glycerine or glucose, cultivate 12h～24h for 30 ℃～37 ℃, again with seed liquor by volume the inoculum size of 1-10% add and to contain glycerine or glucose concn is the initial fermention medium of 30～80g/L, carry out anaerobism or aerobic fermentation under 30～40 ℃ of conditions, feed 0～1.0vvm air during the fermentation, mixing speed 30～350rpm, stream glycerol adding or glucose make in the fermented liquid glycerine or glucose concn be controlled at 10～50g/L; And adopt stream to add 3～4M alkaline solution and control pH value 5.0～8.0; After fermentative production finishes, adopt membrane filtration, centrifugal means with the fermented liquid solid-liquid separation, solid part is a microbial flocculant; When microbial flocculant is used, waste water per ton adds 0～500 gram calcium chloride earlier, adds 0.1～1 kilogram of undried microbial flocculant or the spray-dired microbial flocculant of 20～200 grams again, uses sodium hydroxide adjust pH 4.0～8.0 behind the mixing, mixing again, standing sedimentation.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101941767B (en) * | 2009-07-10 | 2012-03-07 | 中国石油天然气股份有限公司 | Treatment method of 1, 3-propylene glycol concentrated bacterial liquid |
| CN102260729B (en) * | 2011-06-24 | 2013-03-13 | 哈尔滨工业大学 | Bioflocculant fermentation method with mycelium pellet as vector |
| CN103103125B (en) * | 2011-11-10 | 2015-08-12 | 中国石油化工股份有限公司 | A kind of microalgae recovery method that microbial flocculation and air supporting are coupled |
| CN103451232A (en) * | 2012-05-28 | 2013-12-18 | 浙江树人大学 | Preparation method and use method of self-circulation microbial flocculant |
| CN102876600B (en) * | 2012-09-13 | 2013-08-07 | 哈尔滨工业大学宜兴环保研究院 | High-efficiency bioflocculant producing bacterium, screening method thereof and application of high-efficiency bioflocculant producing bacterium in treatment of sulfamethoxazole |
| CN103146797B (en) * | 2013-02-26 | 2015-04-15 | 中国科学院重庆绿色智能技术研究院 | Compound biological flocculant and preparation method thereof |
| CN107381838A (en) * | 2017-08-31 | 2017-11-24 | 常州豪坦商贸有限公司 | A kind of preparation method of compound microbial water purification catridge material |
| CN107794238B (en) * | 2017-11-13 | 2020-10-13 | 中国科学院成都生物研究所 | BFX-01 strain for high yield of biological flocculant and biological flocculant obtained thereby |
| CN112226399B (en) * | 2020-12-11 | 2021-02-26 | 中国科学院上海高等研究院 | Klebsiella (Klebsiella. spp) strain, engineering bacteria and application |
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