CN104694582A - Fermenting method of anaerobe - Google Patents
Fermenting method of anaerobe Download PDFInfo
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- CN104694582A CN104694582A CN201510090875.XA CN201510090875A CN104694582A CN 104694582 A CN104694582 A CN 104694582A CN 201510090875 A CN201510090875 A CN 201510090875A CN 104694582 A CN104694582 A CN 104694582A
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- anaerobion
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/065—Ethanol, i.e. non-beverage with microorganisms other than yeasts
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/16—Butanols
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
- C12P7/26—Ketones
- C12P7/28—Acetone-containing products
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a fermenting method of anaerobe. The method includes at least two steps, wherein in the first step, microorganism is continuously cultured; in the second step, the product is produced by continuous or batch charging, microorganism is fixed on a carrier material; the carrier material is open-celled sintered glass, pumice or equivalents containing lots of SO2 and having open-celled structures; the aperture of the carrier material is 30 to 180 microns, the water absorbing performance is at least 0.35ml/cm<3>, and the ratio of the carrier material to the total operation volume is 1:8 to 1: 1.5. With the adoption of the fermenting method, the output can be stably increased within a long time; in addition, the fermenting method is safe, free of pollution, and high in efficiency, and has a good application prospect.
Description
Technical field
The present invention relates to the fermentation process of a kind of microorganism, particularly relate to the method for carrying out anaerobion fermentation by the microorganism that can produce butanols, acetone, ethanol and/or Virahol, belong to technical field of microbial fermentation.
Background technology
As everyone knows, the cultivation of anaerobion has two traditional culturing steps, namely fermenting process is controlled by through-put or phosphorus restriction hydrochlorate, make to keep constant high concentration microorganism in the first step (cultivation), namely produce stable equilibrium state, and make a part of substrate conversion be butanols, acetone and/or ethanol.Then production peak is reached with product needed for chien shih time short as far as possible at second step.If through-put is too high in the first step, then the cell washed out can be caused more than the new cell grown.On the contrary, when needing can low through-put to control Growth of Cells.
Output still can not be satisfactory first for the shortcoming of the method, second can not keep the permanent stability of cultured continuously, namely can not keep the stable rate of hundreds of hour or longer time.Its reason is that formed product and butanols, acetone and/or ethanol or Virahol have toxic action to organism of fermentation, therefore, when generating product and reaching finite concentration, can stop formation or the fermentation of grievous injury microorganism culturing of product.This currently known methods also another shortcoming noticeable is almost to only have and just can obtains gratifying result with hexose.
Therefore, be badly in need of industry personnel to carry out further exploring and research and development.
Summary of the invention
For the deficiencies in the prior art, the problem to be solved in the present invention is, provides the fermentation process of a kind of anaerobion, and it interiorly for a long time stably can improve output, and safety non-pollution, efficiency is high.
In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:
A fermentation process for anaerobion, the method is carried out at least in two steps, and the first step is cultured continuously microorganism mainly, and second step mainly generates product by continuous or batch charging, at second step by Microorganism incubation on a support material;
Described solid support material adopts open-cell sintered glass, float stone or contains a large amount of SiO
2and there is the equivalent of open-celled structure, aperture is 30-180 um, and water-absorbent is at least 0.35ml/cm
3, solid support material is 1: 8 to 1: 1.5 with the ratio of total operational volume.
Preferably, from product formation stages removing substratum, then by extracting, preferentially adopt currently known methods, be separated with higher alcohols or higher fatty acid the product generated, the more a small amount of product reacting phase of formation is turned back to the first step process of product formation stages and/or present method.
Preferably, when carrying out the first step process, adopting consecutive evaporation method, steaming except product from substratum.
Preferably, adopt evaporating film, especially silicone rubber membrane, steam except product, steaming the temperature removed is 40-80 DEG C, and adopts vector gas, as N
2, H
2, CO
2, CO or these gases mixture, or vapor pres-sure when adopting fermentation gas or adopt vacuum method to reduce evaporation.
Preferably, gas gas stripping process is adopted to carry out best being continuously separated to product in subordinate phase.
Further, stripping gas is adopted, as N
2, H
2, CO
2, CO or these gases mixture, or fermentation gas is continuously separated product, and liquid is 0.06: 1 to 1.8: 1 with the volumetric flow ratio of gas, and temperature is 40-80 DEG C.
Preferably, the add-on of nutritive medium and/or sugar soln is controlled by the product assay of second step process.
Should also be noted in that in a first step the high material throughput of selection number should be able to make microorganism growth, and product acid phase in do not form product significantly.At this growth phase, microorganism develops its good adhesion characteristics, then enters second step with this feature, makes Microorganism incubation on a support material.If the through-put selected in a first step is too low, then cell growth arrest, may product be formed simultaneously, also can lose the microorganism adsorpting characteristic good to solid support material thus.
Adopt microfiltration, be good with crossflow microfiltration method especially, in second step, cell be separated with substratum, Microorganism incubation ground method can be replaced, and then return, so also can obtain the effect same with fixation method.
In addition, high density product (because low through-put causes) can cause the degeneration of microorganism in a first step, thus makes the product of generation generate new acid, and learn from experience, this change is irreversible.And too high in the first step through-put, then microorganism can be washed away, thus after a certain time, cause the cell quantity for second step not enough.
Therefore, through-put must be determined by particular case.
By adopting preceding solution, the invention has the beneficial effects as follows:
Fermentation process of the present invention interiorly for a long time stably can improve output, and safety non-pollution, efficiency is high, and application prospect is good.
Embodiment
Describe embodiments of the present invention in detail below with reference to specific embodiment, to the present invention, how utilisation technology means solve technical problem whereby, and the implementation procedure reaching technique effect can fully understand and implement according to this.
Embodiment 1
The substratum prepared by following composition is used as nutritional medium:
Wood sugar (anhydrous) 60g/l
KH2PO41g/l
K2HPO41g/l
（NH4）2SO42g/l
MgSO4.7H2O 0.1g/l
NaCl 0.01g/l
Na2MoO4.2H2O 0.01g/l
CaCl20.01g/l
MnSO4.H2O 0.015g/l
FeSO4.7H2O 0.015g/l
Vitamin H 0.1mg/l
Thiamine chloride 0.002g/l
Para-amino benzoic acid 0.002g/l
Na2S2O40.035g/l
The reddish black 0.001g/l of cutter
Yeast extract paste 0.5g/l
Na4OOCCH32.2g/l
Clostridium acetobutyricum ATCC824 is used as zymophyte, is used further to the inventive method after first suitably cultivating.
Nutrient medium in gravitation tank is introduced stirred tank bioreactor, and through-put is 0.3/ hour.Inoculum size is about 10% of stirred tank bioreactor capacity, ferments under remaining on 37 DEG C of temperature and pH5.1 in the reactor.Cell density in bio-reactor roughly remains unchanged by selecting through-put, fixed-bed reactor are entered by pipeline so as to making the new cell by producing in microorganism culturing, due to cell adhesion on a support material, so they still keep adherence state in the reactor.Adopt open-cell sintered glass as solid support material, the volume of sintered glass is equivalent to 50% of cumulative volume.Temperature in fixed-bed reactor remains on 43 DEG C, and pH4.3 ferments.In two-step pretreatment, total through-put is 0.15/ hour.Reactant nitrogen in fixed-bed reactor is inflated, because fixed-bed reactor are built with sleeve pipe, so can keep circulation.
Productive rate and output provide in Table 1.
Table 1
Thinning ratio (hour-1) 0.15
Flow out wood sugar (g/l) 28.59
Sugar utilization (%) 50.98
Sugar utilization (g/lhr) 4.46
Butanols (g/l) 5.7
Total solvent amount (g/l) 9.30
Solvent production (%) 31.28
Solvent productive rate (g/lhr) 1.395
Total acid content (g/l) 3.06
Acid yield (%) 5.88
Optical density(OD) 2.765
pH 4.32
Electronegativity (mv)-436
Embodiment 2
Identical nutritive compositions, inoculation and method condition is selected in stirred tank bioreactor and fixed-bed reactor.One therefrom can be obtained continuously or in batches the outer circulation separation system generating product to be connected with fixed-bed reactor.In addition, after the substratum being rich in product obtained is heated to about 40 DEG C, be incorporated into stripping tower by fixed-bed reactor, to introduce in tower nitrogen as stripping gas with liquid countercurrent direction.Liquid is 0.35: 1 with the volumetric flow ratio of gas.Substantially the substratum of product crossed by stripping, and wherein a part enters into fixed-bed reactor again after being cooled to leavening temperature, and small portion also can turn back to stirred tank bioreactor.
Substantially the remaining liq that product crossed by stripping is got rid of from system.The gas stream being rich in product of discharging from stripping tower, through cooling system, is discharged after product cohesion.The steady-state value obtained provides in table 2.
Table 2
Thinning ratio (hour-1) 0.15
Flow out wood sugar (g/l) 21.50
Sugar utilization (%) 62.50
Sugar utilization (g/lhr) 5.37
Butanols (g/l) 6.82
Total solvent amount (g/l) 10.87
Solvent production (%) 30.36
Solvent productive rate (g/lhr) 1.638
From table, employing embodiment 2 method, namely from the product that outer circulation separation system obtains continuously, its solvent productive rate and sugared utilization obviously better.As shown in above-mentioned experiment, if fermentation is for a long time as 500 hours, also effect same can be obtained.
In sum, fermentation process of the present invention interiorly for a long time stably can improve output, and safety non-pollution, efficiency is high, and application prospect is good.
Above-mentioned embodiment; be only and technical conceive of the present invention is described; object is to allow the stakeholder being familiar with technique implement according to this; but above said content does not limit the scope of the invention; every any equivalence done according to spirit of the present invention changes or modifies, and all should fall within protection scope of the present invention.
Claims (7)
1. a fermentation process for anaerobion, the method is carried out at least in two steps, and the first step is cultured continuously microorganism mainly, and second step mainly generates product by continuous or batch charging, it is characterized in that, at second step by Microorganism incubation on a support material;
Described solid support material adopts open-cell sintered glass, float stone or contains a large amount of SiO
2and there is the equivalent of open-celled structure, aperture is 30-180 um, and water-absorbent is at least 0.35ml/cm
3, solid support material is 1: 8 to 1: 1.5 with the ratio of total operational volume.
2. the fermentation process of anaerobion according to claim 1, it is characterized in that, from product formation stages removing substratum, then by extracting, preferential employing currently known methods, be separated with higher alcohols or higher fatty acid the product generated, the more a small amount of product reacting phase of formation is turned back to the first step process of product formation stages and/or present method.
3. the fermentation process of anaerobion according to claim 2, is characterized in that, when carrying out the first step process, adopting consecutive evaporation method, steaming except product from substratum.
4. the fermentation process of anaerobion according to claim 3, is characterized in that, adopts evaporating film, especially silicone rubber membrane, and steam except product, steaming the temperature removed is 40-80 DEG C, and adopts vector gas, as N
2, H
2, CO
2, CO or these gases mixture, or vapor pres-sure when adopting fermentation gas or adopt vacuum method to reduce evaporation.
5. the fermentation process of anaerobion according to claim 1, is characterized in that, adopts gas gas stripping process to carry out best being continuously separated to product in subordinate phase.
6. the fermentation process of anaerobion according to claim 4, is characterized in that, adopts stripping gas, as N
2, H
2, CO
2, CO or these gases mixture, or fermentation gas is continuously separated product, and liquid is 0.06: 1 to 1.8: 1 with the volumetric flow ratio of gas, and temperature is 40-80 DEG C.
7. the fermentation process of anaerobion according to claim 1, is characterized in that, controls the add-on of nutritive medium and/or sugar soln by the product assay of second step process.
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| CN201510090875.XA CN104694582A (en) | 2015-02-28 | 2015-02-28 | Fermenting method of anaerobe |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN88101266A (en) * | 1987-03-10 | 1988-09-28 | 赫尔穆特·埃芬堡格 | The microbial fermentation processes that contains culture medium of carbohydrate |
| CN1228124A (en) * | 1994-11-30 | 1999-09-08 | 生物工程资源股份有限公司 | Biological production of acetic acid from waste gases |
| CN1444658A (en) * | 2000-07-25 | 2003-09-24 | 生物工程资源股份有限公司 | Methods for increasing production of ethanol from microbial fermentation |
| CN102676589A (en) * | 2012-05-09 | 2012-09-19 | 大连理工大学 | Method for producing, separating and purifying butanol by coupling fermenting with gas stripping |
| CN103555560A (en) * | 2013-10-28 | 2014-02-05 | 南京工业大学 | Device and method for preparing butanol through fermentation, coupling, separation and purification of acetone-butanol |
-
2015
- 2015-02-28 CN CN201510090875.XA patent/CN104694582A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN88101266A (en) * | 1987-03-10 | 1988-09-28 | 赫尔穆特·埃芬堡格 | The microbial fermentation processes that contains culture medium of carbohydrate |
| CN1228124A (en) * | 1994-11-30 | 1999-09-08 | 生物工程资源股份有限公司 | Biological production of acetic acid from waste gases |
| CN1444658A (en) * | 2000-07-25 | 2003-09-24 | 生物工程资源股份有限公司 | Methods for increasing production of ethanol from microbial fermentation |
| CN102676589A (en) * | 2012-05-09 | 2012-09-19 | 大连理工大学 | Method for producing, separating and purifying butanol by coupling fermenting with gas stripping |
| CN103555560A (en) * | 2013-10-28 | 2014-02-05 | 南京工业大学 | Device and method for preparing butanol through fermentation, coupling, separation and purification of acetone-butanol |
Non-Patent Citations (3)
| Title |
|---|
| SI-YU LI 等: "Performance of batch, fed-batch, and continuous A–B–E fermentation with pH-control", 《BIORESOURCE TECHNOLOGY》 * |
| 王鑫昕 等: "发酵罐水平的气提-萃取-发酵生产丁醇", 《沈阳农业大学学报》 * |
| 郭加明 等: "乙醇补料发酵技术研究进展", 《化工进展》 * |
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