CN103060391A - Low-temperature sterilization method suitable for cellulosic ethanol production - Google Patents
Low-temperature sterilization method suitable for cellulosic ethanol production Download PDFInfo
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- CN103060391A CN103060391A CN201310014119XA CN201310014119A CN103060391A CN 103060391 A CN103060391 A CN 103060391A CN 201310014119X A CN201310014119X A CN 201310014119XA CN 201310014119 A CN201310014119 A CN 201310014119A CN 103060391 A CN103060391 A CN 103060391A
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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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- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/59—Biological synthesis; Biological purification
Abstract
The invention discloses a low-temperature sterilization method suitable for cellulosic ethanol production. The low-temperature sterilization method comprises the following steps of: (1) directly adding weighed quantitative cellulose material subjected to pretreatment together with other elements of a culture medium into a fermentation kettle; (2) adding quantitative cellulase solution according to the final fermentation volume, heating up to 50-65 DEG C, preserving for 1-3 hours, and then cooling to a fermentation temperature of ethanol fermentation strains; and (3) adding cultured ethanol fermentation strain seed suspension by a sterile inoculation way, keeping the pressure of the kettle at 0.01-0.02MPa, and controlling an appropriate condition to carry out anaerobic fermentation till finishing the fermentation. The low-temperature sterilization method disclosed by the invention has the characteristics that impure bacteria are not mixed and the catalytic efficiency of the cellulase is considered at the same time, suppressive products produced by a traditional sterilization method are eliminated, the energy consumption is reduced, and the sterilization cost is saved.
Description
Technical field
The invention belongs to the cellulose alcoholic fermentation field, specifically, relate to a kind of temperature sterilization method that is applicable to cellulosic ethanol production.
Background technology
Along with petering out of fossil energy, the appearance of the phenomenons such as the climate warming that the consumption fossil energy is brought, environmental degradation, the hope that people are urgent finds a kind of reproducible clean energy as the substitute of fossil energy, reduces environmental disruption when satisfying consumption requirements.Alcohol fuel is as a kind of reproducible clean energy, carry out in the U.S., Brazil since the seventies in last century, because the initial alcohol fuel great majority of carrying out are to be raw material by corn and other starches matter food crop, cause the rise of world food price, form the situation of striving grain with the people, restricted its large-scale promotion.Along with the development of cellulase technology, the exploitation of cellulosic ethanol technology and cellulase are commercially produced and are just gradually made good and scale.
In the production strategy of cellulosic ethanol, synchronous saccharification and fermentation technique (SSF) are considered to both economical, and can save fermentation time, the inhibition of elimination product, are fit to large-scale promotion.By simultaneously dropping in fermentor tank in the fermentation starting stage cellulosic material, ethanol fermentation bacterial strain and cellulase, the ethanol fermentation bacterial strain consumes the glucose of generation when the cellulose degraded Mierocrystalline cellulose generates glucose, so that the content of glucose maintains a very low concentration in the whole fermenting process, reduced greatly because the enzymolysis that the accumulation of glucose causes suppresses phenomenon, improved the efficient of enzymolysis; Also reduce simultaneously glucose and be used in a large number the generation that the fermentation strain growth has brought the problem of transformation efficiency decline.
For to guarantee that fermenting process does not have the pollution of external source bacterium, fermention medium will carry out sterilising treatment by wet-hot steam in advance in synchronous saccharification and fermentation technique, to eliminate the external source miscellaneous bacteria.Then the enzyme liquid and the good fermentation strain of inoculation culture that add degerming.This kind sterilising method exists some drawbacks: 1. lignocellulosic material easily generates furfural, polyphenolic substance under 115 ~ 121 ℃ wet-hot steam sterilization, such material is the inhibitor of cellulase, suppress greatly cellulase in cellulosic combination, reduce enzymolysis efficiency; 2. the concentration that will guarantee final ethanol in the high density cellulose alcoholic fermentation process improves the concentration that must improve the cellulosic material in early stage, because cellulosic material mostly is water-fast solid matter, a large amount of water of condensation can cause fermentation volume to become large final impact fermentation concentration during the larger wet-hot steam sterilization of shared volume ratio; 3. affect heat transfer owing to can cause fermented liquid to become sticky under the cellulose raw material high density, thereby when affecting fermentation volume, also can cause the low of heat utilization ratio, cause the sterilization energy consumption excessive, improved greatly cost; Since the general the suitableeest enzyme work of cellulase about 50 ℃, and present high temperature resistant fermentation strain is generally at 30 ~ 40 ℃, there is the suitableeest contradiction, for being chosen under the lower temperature of cellulase catalytic efficiency, the survival that guarantees fermentation strain ferments, reduced greatly the catalytic efficiency of cellulase, so that fermentation period prolongs.
Summary of the invention
The objective of the invention is: a kind of temperature sterilization method that is applicable to cellulosic ethanol production is provided, deficiency for above 4 synchronous saccharifications and fermentation technique, this temperature sterilization method is taken into account the catalytic efficiency of cellulase when guaranteeing not dye miscellaneous bacteria, eliminate the inhibition product that traditional sterilization method produces, reduce the energy consumption utilization, save the sterilization cost.
Technical solution of the present invention is that this temperature sterilization method may further comprise the steps:
(1) other components that take by weighing the pretreated cellulosic material of quantitative process and substratum directly add in the fermentor tank;
(2) add quantitative cellulose enzyme liquid and be warming up to 50 ~ 65 ℃ according to final fermentation volume, be incubated 1 ~ 3h, be cooled to the leavening temperature of ethanol fermentation bacterial strain;
(3) mode by aseptic inoculation adds cultured ethanol fermentation bacterial strain seed suspension, keeps tank pressure at 0.01 ~ 0.02Mpa, controls suitable condition and carries out anaerobically fermenting until fermentation ends.
Wherein, the source of the cellulosic material in the step 1) is cornstalk, the straw of reproducible wood fiber biomass, or is the relevant raw material of cellulosic that papermaking, textile industry produce.
Wherein, the pretreatment mode of the cellulosic material in the step 1) is chemically treated acid treatment, alkaline purification, or the Irradiation of physical treatment, mechanical treatment, or biological treatment and gas explosion process, or is the combined treatment of several processing modes.
Wherein, step 2) the final fermentation volume of the basis in adds cellulose enzyme liquid and refers to the cellulose enzyme amount required according to fermentation, measure or take by weighing quantitative enzyme powder or enzyme liquid, be diluted with water to the final fermentation volume place of deduction inoculation volume, be pressed in the fermentor tank cellulose raw material and other nutritive ingredient mixings with early investment, or directly being pressed into volume is the fermentation crude enzyme liquid that final fermentation volume deducts the seed volume.
Wherein, step 2) cellulase described in is the bacterium of one or more cellulase-producings or enzyme liquid, the enzyme powder that fungi fermentation obtains, and comprises endoglucanase, exoglucanase and beta-glucosidase.
Wherein, the keeping warm mode step 2) need to be opened and stir to accelerate the material mixing.
The present invention compares with tradition following advantage: 1. without high-temp steam sterilizing, thereby do not have corresponding enzyme to suppress the product generation; 2. can kill other bacterium except thermophile bacteria or genus bacillus etc. by cellulose raw material and enzyme 50 ~ 60 ℃ insulation, the insulation of short period of time is so that the enhancing of cellulose degradation flow of solution simultaneously, increased the mass-transfer efficiency of solution, fermention medium after the insulation contains the monose of a large amount of cellulose degradations, behind access ethanol fermentation bacterial strain, can guarantee flora amplification in the early stage nutritional needs of bacterial strain, form the flora advantage in the short period of time and suppress varied bacteria growing, short period of time produces great amount of carbon dioxide reduction medium pH simultaneously, and form the sour environment that the carbonic acid gas barrier film forms a complete anaerobism at the fermentor tank top, by the flora advantage, the measures such as complete anaerobic environment have suppressed the growth of external source miscellaneous bacteria, have formed the effect of pure culture; 3. the enzyme liquid that adds can be aseptically process, also can be not have treated enzyme liquid simultaneously, has reduced the cost of enzyme liquid aseptically process; 4. greatly reduce energy consumption, shortened simultaneously the time of cellulose degradation.
Embodiment
Below by specific embodiment content of the present invention is described in detail, these embodiment can not be interpreted as it is restriction to technical scheme.
Embodiment 1: according to the following steps production of cellulosic ethanol
(1) cellulosic material: straw, humidity about 20%, mean length are about 2 centimetres; Pre-treatment is directly 2 kilograms of straw to be placed 50 liters pressurized vessel, processes 20 minutes post-dryings under 200 ℃ wet-hot steam;
(2) substratum: take by weighing pretreated straw 15kg, (NH
4)
2SO
4300g, KH
2PO
4300g, MgSO
445g and CSL 300g add in the 250L fermentor tank, add to be warming up to 121 ℃ of insulation 20min after water 70L stirs and to be cooled to 37 ℃;
(3) enzyme-added liquid and inoculation: being pressed into the total enzyme work of cellulose enzyme liquid 23L filter paper enzyme activity of processing through degerming by aseptic culture transferring pipeline is 7.5 ten thousand IU(international unit) cellulase solution, and the culture transferring pipeline by sterilization in advance cultured volume be that the 7L fermented incubation time is that the high temperature resistant yeast saccharomyces cerevisiae of 10h is pressed in the ethanol fermentation tank;
(4) zymamsis is cultivated: control tank temperature is 37 ℃, stirs to be 200rpm/min, and regularly fermentor tank exhaust maintenance fermentor tank tank pressure is 0.01Mpa, the 5 days fermentation ends of fermenting.
Fermentation results: ethanol concn is 16g/L in the fermented liquid through measuring.
Embodiment 2: according to the following steps production of cellulosic ethanol
(1) cellulosic material: straw, humidity are about 20%, and mean length is about 2 centimetres; Pre-treatment is directly 2 kilograms of straw to be placed 50 liters pressurized vessel, processes 20 minutes post-dryings under 200 ℃ wet-hot steam;
(2) substratum: take by weighing pretreated straw 15kg, (NH
4)
2SO
4300g, KH
2PO
4300g, MgSO
445g and CSL 300g add in the 250L fermentor tank;
(3) cellulase powder 250g enzyme-added liquid: take by weighing filter paper enzyme activity and be the enzyme that the 300IU/g(international unit represents and live) dissolves with the tap water of pH4.8 and is pressed in the fermentor tank after being dissolved to the 93L mixing, open to stir and be 200rpm/min, be cooled to 37 ℃ after being warming up to 50 ℃ of insulation 3h;
(4) inoculation: by the culture transferring pipeline of sterilization will be in advance cultured volume be that the 7L fermented incubation time is that the high temperature resistant yeast saccharomyces cerevisiae of 10h is pressed in the ethanol fermentation tank;
(5) zymamsis is cultivated: control tank temperature is 37 ℃, stirs to be 200rpm/min, and regularly fermentor tank exhaust maintenance fermentor tank tank pressure is 0.01Mpa, cultivates 3 days fermentation ends.
Fermentation results: ethanol concn is 19g/L in the fermented liquid through measuring.
Embodiment 3: according to the following steps production of cellulosic ethanol
(1) cellulosic material: maize straw, humidity are about 20%, and mean length is about 2 centimetres; Pre-treatment is directly 2 kilograms of stalks to be placed 50 liters pressurized vessel, adds mass concentration 2% NaOH 20L at 90 ℃ of insulation 4h, collects solid straws with washing from the beginning post-drying 3 times;
(2) substratum: take by weighing pretreated stalk 15kg, (NH
4)
2SO
4300g, KH
2PO
4300g, MgSO
445g and CSL 300g add in the 250L fermentor tank;
(3) cellulase solution 2.5L enzyme-added liquid: measure filter paper enzyme activity and be the enzyme that the 30IU/ml(international unit represents and live) is pressed in the fermentor tank after being dissolved to the 93L mixing with the tap water of pH4.8, open to stir and be 200rpm/min, be cooled to 37 ℃ after being warming up to 60 ℃ of insulation 2h;
(4) inoculation: by the culture transferring pipeline of sterilization will be in advance cultured volume be that the 7L fermented incubation time is that the high temperature resistant yeast saccharomyces cerevisiae of 10h is pressed in the ethanol fermentation tank:
(5) zymamsis is cultivated: control tank temperature is 37 ℃, stirs to be 200rpm/min, and regularly fermentor tank exhaust maintenance fermentor tank tank pressure is 0.015Mpa, cultivates 4 days fermentation ends.
Fermentation results: ethanol concn is 17g/L in the fermented liquid through measuring.
Embodiment 4: according to the following steps production of cellulosic ethanol
(1) cellulosic material: the waste paper of industrial waste;
(2) substratum: take by weighing industrial waste waste paper 10kg, (NH
4)
2SO
4300g, KH
2PO
4300g, MgSO
445g and CSL 300g add in the 250L fermentor tank;
(3) enzyme-added liquid: be pressed into the fermented liquid 93L after the Trichodermareesei RUTC-30 fermentation ends in the fermentor tank, total filter paper enzyme activity is the international enzyme of the 150000 IU(unit that lives), open to stir and be 200rpm/min, be cooled to 37 ℃ after being warming up to 65 ℃ of insulation 1h;
(4) inoculation: by the culture transferring pipeline of sterilization will be in advance cultured volume be that the 7L fermented incubation time is that the high temperature resistant yeast saccharomyces cerevisiae of 10h is pressed in the ethanol fermentation tank;
(5) zymamsis is cultivated: control tank temperature is 37 ℃, stirs to be 200rpm/min, and regularly fermentor tank exhaust maintenance fermentor tank tank pressure is 0.02Mpa, cultivates the 80h secondary fermentation and finishes.
Fermentation results: ethanol concn is 33g/L in the fermented liquid through measuring.
The method by temperature sterilization of can finding out from above embodiment can guarantee carrying out smoothly of fermenting process, can save time greatly and energy consumption simultaneously.
Claims (6)
1. be applicable to the temperature sterilization method of cellulosic ethanol production, it is characterized in that this temperature sterilization method may further comprise the steps:
(1) other components that take by weighing the pretreated cellulosic material of quantitative process and substratum directly add in the fermentor tank;
(2) add quantitative cellulose enzyme liquid and be warming up to 50 ~ 65 ℃ according to final fermentation volume, be incubated 1 ~ 3h, be cooled to the leavening temperature of ethanol fermentation bacterial strain;
(3) mode by aseptic inoculation adds cultured ethanol fermentation bacterial strain seed suspension, keeps tank pressure at 0.01 ~ 0.02Mpa, controls suitable condition and carries out anaerobically fermenting until fermentation ends.
2. the temperature sterilization method that is applicable to cellulosic ethanol production according to claim 1, it is characterized in that: wherein, the source of the cellulosic material in the step 1) is cornstalk, the straw of reproducible wood fiber biomass, or is the relevant raw material of cellulosic that papermaking, textile industry produce.
3. the temperature sterilization method that is applicable to cellulosic ethanol production according to claim 1, it is characterized in that: wherein, the pretreatment mode of the cellulosic material in the step 1) is chemically treated acid treatment, alkaline purification, or the Irradiation of physical treatment, mechanical treatment, or biological treatment and gas explosion processing, or be the combined treatment of several processing modes.
4. the temperature sterilization method that is applicable to cellulosic ethanol production according to claim 1, it is characterized in that: wherein, step 2) the final fermentation volume of the basis in adds cellulose enzyme liquid and refers to the cellulose enzyme amount required according to fermentation, measure or take by weighing quantitative enzyme powder or enzyme liquid, be diluted with water to the final fermentation volume place of deduction inoculation volume, be pressed in the fermentor tank cellulose raw material and other nutritive ingredient mixings with early investment, or directly being pressed into volume is the fermentation crude enzyme liquid that final fermentation volume deducts the seed volume.
5. the temperature sterilization method that is applicable to cellulosic ethanol production according to claim 1, it is characterized in that: wherein, step 2) cellulase described in is the bacterium of one or more cellulase-producings or enzyme liquid, the enzyme powder that fungi fermentation obtains, and comprises endoglucanase, exoglucanase and beta-glucosidase.
6. the temperature sterilization method that is applicable to cellulosic ethanol production according to claim 1 is characterized in that: wherein, step 2) in keeping warm mode need to open and stir to accelerate the material mixing.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337936A (en) * | 2018-10-30 | 2019-02-15 | 孙金良 | Association fibre element Ethanol Method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1205743A1 (en) * | 2000-06-29 | 2002-05-15 | Hogy Medical Co., Ltd. | Indicator for plasma sterilization |
| CN1807610A (en) * | 2006-01-23 | 2006-07-26 | 迟乃玉 | Method for producing low temperature cellulase using microbe fermentation |
| CN102174410A (en) * | 2009-10-22 | 2011-09-07 | 郑州大学 | Stain for producing heat-resisting and acid-resisting cellulase with high yield and new method for fermenting cellulosic ethanol |
| CN102776242A (en) * | 2011-05-13 | 2012-11-14 | 国家海洋局第一海洋研究所 | Process method for producing ethanol by utilizing low-temperature cellulase to perform simultaneous saccharification and fermentation |
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2013
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1205743A1 (en) * | 2000-06-29 | 2002-05-15 | Hogy Medical Co., Ltd. | Indicator for plasma sterilization |
| CN1807610A (en) * | 2006-01-23 | 2006-07-26 | 迟乃玉 | Method for producing low temperature cellulase using microbe fermentation |
| CN102174410A (en) * | 2009-10-22 | 2011-09-07 | 郑州大学 | Stain for producing heat-resisting and acid-resisting cellulase with high yield and new method for fermenting cellulosic ethanol |
| CN102776242A (en) * | 2011-05-13 | 2012-11-14 | 国家海洋局第一海洋研究所 | Process method for producing ethanol by utilizing low-temperature cellulase to perform simultaneous saccharification and fermentation |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337936A (en) * | 2018-10-30 | 2019-02-15 | 孙金良 | Association fibre element Ethanol Method |
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Inventor after: Zhao Jiehong Inventor after: Xiong Peng Inventor after: He Jianlong Inventor after: Yuwen Weigang Inventor after: Xu Jiming Inventor before: Xiong Peng Inventor before: He Jianlong Inventor before: Yuwen Weigang Inventor before: Xu Jiming |
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Free format text: CORRECT: INVENTOR; FROM: XIONG PENG HE JIANLONG YUWEN WEIGANG XU JIMING TO: ZHAO JIEHONG XIONG PENG HE JIANLONG YUWEN WEIGANG XU JIMING |
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Application publication date: 20130424 Assignee: Huaian Bama Keyu green biological energy Co. Ltd. Assignor: Xiong Peng Contract record no.: 2016320000228 Denomination of invention: Low-temperature sterilization method suitable for cellulosic ethanol production Granted publication date: 20141217 License type: Exclusive License Record date: 20161226 |
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