CN104372032A - Method for producing fuel ethanol by fermenting wine lees - Google Patents
Method for producing fuel ethanol by fermenting wine lees Download PDFInfo
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- CN104372032A CN104372032A CN201410639840.2A CN201410639840A CN104372032A CN 104372032 A CN104372032 A CN 104372032A CN 201410639840 A CN201410639840 A CN 201410639840A CN 104372032 A CN104372032 A CN 104372032A
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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
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
- C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
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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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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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 Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
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- Bioinformatics & Cheminformatics (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for producing fuel ethanol by fermenting wine lees. The method disclosed by the invention can be used for voluntarily preparing a complex enzyme system, namely high temperature-resistant alpha-amylase, saccharifying enzyme, cellulase and acid protease, can be used for greatly enhancing the effect of decomposing and cutting capacity and effectively decomposing cellulose, xylogen, crude starch, crude protein and pectin in the wine lees, and can be used for solving the problem of pollution of solid wastes on the environment and increasing the demand of energy fuel of a company by producing fuel ethanol by using the wine lees and well solving the problem of organic combination of the cellulose, acid hydrolysis and enzyme hydrolysis and transforming the biological substances into ethanol again by continuously carrying out non-isothermal simultaneous liquefaction, saccharification and fermentation in a same container. According to the method, residues generated in a production process can be taken as feeds for developing the animal husbandry, and microbes are fermented to produce biogas, organic fertilizers and planting edible fungi for developing the planting industry.
Description
Technical field
The present invention relates generally to fuel area, particularly relates to a kind of method that bent vinasse fermentation produces alcohol fuel.
Background technology
The bent vinasse of a large amount of wastes can be produced in bent wine production process, although and these wastes can as the feed a part source of some domestic animal, because its content of cellulose is high, therefore nutritive value is lower, can only utilize a part, and major part still does offal treatment; In recent years, due to the development of science and technology and the raising of feeding quality level, the trophic structure in vinasse can not the requirement of full feeding industry scientific feeding and the needs of scale development.Open up new vinasse to utilize, become the focus of liquor industry.For the practical situation that above reason and bent wine are produced, the thinking using bent vinasse as novel biomass raw material production ethanol emerges.Alcohol fuel is a kind of clean, the reproducible energy of alternative oil in the world at present, and the solid waste of liquor-making enterprises-bent vinasse processing costs is high, is something that Liquor-making Enterprises & compares headache.
Summary of the invention
Object of the present invention is with fermentation process, utilizes various composition in the bent vinasse of combinative enzyme hydrolysis, generates novel energy alcohol fuel, realize the comprehensive utilization of by product, solve the pollution problem of solid waste to environment.
The present invention is achieved by the following technical solutions:
Bent vinasse fermentation produces a method for alcohol fuel, comprises the following steps:
(1) vinasse 480-500g is got, dry, pulverize, cross 0.8-1.0mm sieve plate hole, be placed in 3L fermentation flask, the warm water adding 1400-1500ml, 32-34 DEG C is sized mixing, and adds urea 13-15g, ammonium sulfate 10-15g, drips sulfuric acid after stirring, regulate PH to 3.5-4.0, with three layers of gauze wrapping sealing;
(2) put into steam sterilizer, at 0.1-0.105MPa, 120-122 DEG C, carry out high-temperature sterilization 110-130 minute, step-down is lowered the temperature, take out, when being cooled to 48-50 DEG C to temperature, the prozyme adding system weight of material 1.8-2.5% be hydrolyzed, liquefy, saccharification process, described prozyme is the Thermostable α-Amylase of 1:1:1:0.5-1 by mass ratio, saccharifying enzyme, cellulase, aspartic protease mixing composition, process and be cooled to 31-33 DEG C after 60-65 minute, add the aspartic protease of above-mentioned equal quality and the dry yeast activation solution of resurrection, the dry yeast activation solution of described resurrection is by yeast temperature 35-40 DEG C, the syrup rehydration 15-20 minute of concentration 2-3%, activation preparation in 1-2 hour when being then cooled to 32-33 DEG C, the inoculum size of the dry yeast activation solution brought back to life is the 12-15% of system weight of material, be placed in the cyclotron oscillation device shaking flask constant temperature culture of 100r/min in sterilisable chamber, 8-10h is cultivated with two-layer felt or three layers of gauze sealing, sampling microscopy yeast count and bud ratio, then the aseptic biochemical cultivation case putting into 32-36 DEG C ferments, by 4h, 8h, 12h respectively vibrates and stirs once, stir once every 7-8h later, middle and later periods fermentation is entered after 40h, stir once every 12h, 72h fermentation ends, take out fermenting-ripening wine with dregs, both alcohol fuel was obtained again by distillation.
Advantage of the present invention is:
1, prepare multiply anchor-pile voluntarily, i.e. Thermostable α-Amylase, saccharifying enzyme, cellulase, aspartic protease, the effect of decomposing cutting power substantially increases, and effectively the Mierocrystalline cellulose in vinasse, xylogen, Crude starch, crude protein, pectin is decomposed.
2, produce alcohol fuel with bent vinasse, both solved the pollution of solid waste to environment, increase again the demand of company's energy fuel simultaneously.
3, in same container, carry out that non-isothermal liquefies simultaneously continuously, saccharification, fermentation.Solving Mierocrystalline cellulose well, acid hydrolysis is organically combined with enzymic hydrolysis, is ethanol by Wood Adhesives from Biomass again.
4, the slag of production process generation, can as feed advance livestock industry, and fermentable produces biogas, fertilizer, plantation edible mushrooms, development plant husbandry.
5, boiling temperature controls not high, and within the scope of 120-122 DEG C, and Alcohol Production boiling temperature requires at 136-138 DEG C, greatly reduces steam consumption.
6, this technology can make use of the Multiple components in solid waste-bent vinasse well, and it is better to go out rate, produces 95.5 ° of ethanol of 24%.
7, the rice husk in vinasse is through repeatedly repeating fermented soy, pyrogenic distillation, chaff shell softening fibre element, hemicellulose add sulfuric acid through pre-treatment again and adjust pH value, pyrohydrolysis, can effectively destroy cellulosic crystalline structure, make chaff shell become loose hydrolyzed hemicellulose effectively, thus chaff shell is fully used
Accompanying drawing explanation
Fig. 1 is ethanol fermentation curve of the present invention;
Embodiment
Embodiment 1
Bent vinasse fermentation produces a method for alcohol fuel, comprises the following steps:
Vinasse conventional sense composition result unit g/100g
1, bent vinasse pre-treatment:
Vinasse are dried and are pulverized, and as in 3L fermentation flask, reach thick mash fermentation technique Raw and pulverize profit slurry concentration requirement.Stir evenly simultaneously, put into steam sterilizer and carry out step-down after pyrohydrolysis.Cooling taking-up is cooled to 50 DEG C and adds prozyme and carry out enzymatic saccharification.
2, complex enzyme hydrolysis, liquefaction, saccharification:
The synergy of enzyme, just as " relay race ", such as Crude starch generates a large amount of dextrin under the effect of Thermostable α-Amylase.Need be decomposed by saccharifying enzyme.Mierocrystalline cellulose, need be hydrolyzed by cellulase.Protein need be degraded by aspartic protease.Otherwise affect semi-manufactured goods quality, fermenting speed and the yield of liquor.
Each enzyme just can only can show maximum enzyme activity within the scope of certain pH value, i.e. the optimum pH of enzyme.The optimal pH of prozyme is 3.5 – 4.0.The optimum pH of prozyme by adding when moistening slurry that sulfuric acid adjusts between PH to 3.5 – 4.0.Further raising enzyme decomposition reaction velocity.Shorten saccharification time, improve enzyme utilization ratio.
Enzymic activity in saccharification to raising Crude starch and cellulose utilization rate very important.Due to prozyme decomposition, liquefaction, saccharification, whether thoroughly the large young pathbreaker of consumption is one and is directly connected to saccharification react key issue.
Prozyme is to the vegetable cell in raw material, and fiber boundary dextrin and remaining starch produce hydrolysis, liquefaction, saccharification are strong.Prozyme optimum temperuture is 48 – 52 DEG C, action time about 60min.Under this condition, enzyme decomposition reaction is the suitableeest.Hydrolysis, liquefaction, saccharification can be given full play to.Can use with several enzyme simultaneously add, therefore enzyme has specificity, does not interfere with each other.Simultaneously due to the synergy of multi-enzyme system, its effect is better.
The performance characteristics of prozyme:
1, cellulase is a kind of prozyme, primarily of circumscribed β-glucose, and the compositions such as inscribe β-glucolase and β-Isosorbide-5-Nitrae-glucoside bond.Mierocrystalline cellulose is made to become cellobiose and glucose.Natural cellulose can be decomposed into short chain cellulose, by straight chain Mierocrystalline cellulose inner cut-out, glucose molecule oligosaccharides and molecular weight polymers can be resolved into.Carboxymethyl cellulose can be hydrolyzed to cellobiose, and cellobiose is hydrolyzed to glucose.It also can destroy plant cell wall.Lysis is fully utilized.Promote starch release and cellulosic brief introduction, improve fermentation rate.
2, Thermostable α-Amylase acts on amylose starch, and after α-Isosorbide-5-Nitrae-glycosidic link is cut into short chain dextrin brokenly, dextrin is continued enzymolysis and inoperative to cellulosic β-Isosorbide-5-Nitrae-glycosidic link.It also acts on amylopectin (also known as amylan), cuts α-Isosorbide-5-Nitrae-glycosidic link equally, can not cut α-1.6-glycosidic link.
3, the reducing end under neutral glucose unit from starch can cut down by saccharifying enzyme one by one, generates glucose.Simultaneously again can tapping point in hydrolyzing amylopectin.Generally α-Isosorbide-5-Nitrae-key is disconnected, then continue hydrolysis incision α-1,6 – glycosidic link and α-1,3 – glycosidic link.Finally amylose starch and amylopectin all can be hydrolyzed to glucose.
4, aspartic protease: this enzyme is similar to the stomach en-of animal and the character of rennin.The optimal pH of this enzyme is between 2.5 – 5.5.When PH raises, enzyme activity disappears very soon.Optimum temperuture is 30 – 45 DEG C.Enzyme deactivation when temperature raises.The protein of this enzyme in the abundant hydrolysis material of Crude starch earlier fermentation, improves yeast absorbability nitrogen, a-amino acid, carbon source, yeast rate of propagation is accelerated.
5, the synergy of enzyme, just as " relay race ", such as Crude starch generates a large amount of dextrin under the effect of Thermostable α-Amylase.Need be decomposed by saccharifying enzyme.Mierocrystalline cellulose, need be hydrolyzed by cellulase.Protein need be degraded by aspartic protease.Otherwise affect semi-manufactured goods quality, fermenting speed and the yield of liquor.
Pretty youth is lived according to good fortune in 1979, soluble components is become under the effect of the phenol oxidase (laccase) that xylogen can produce in microorganism, again under the effect of the relevant oxidases of cytopigment, generate the products such as forulic acid, Vanillin, vanillic acid, coumaric acid further.All can be undertaken by yeast and fermentation using bacteria.Therefore material change is complicated, that it is hard to tell is perfectly clear.
6, enzymatic reaction is generally all carried out under the conditions such as comparatively gentle temperature, pH value, and the requirement for conversion unit is also lower.Now understand multiply anchor-pile acting in conjunction Crude starch, Mierocrystalline cellulose, breaks down proteins are transformed into glucose yeast and glucose could be become ethanol.
3, the ethanol fermentation of vinasse acid hydrolysis and enzyme hydrolyzate:
1, aerobic incubation period temperature 31 – 33 DEG C, time 8 – 10h.After acid hydrolysis and enzyme hydrolyzate access yeast, be placed in constant temperature oxygen supply in the shaking flask in the cyclotron oscillation device of 31 – 33 DEG C, 100r/min in sterilisable chamber and cultivate.Promote the breeding of yeast, cytoactive is strengthened, density increases.Make fermented liquid stir enhancing, facilitate acid hydrolysis and enzymic hydrolysis.Reach the parallel binary catalysts method carried out fermenting in enzymatic saccharification limit, limit.Fermentation reaches best effect, improves the utilization ratio of Mierocrystalline cellulose and residual starch.Reduce Yeast Flocculation, discharge more yeast cell and participate in reaction.Improve the contact surface area of yeast, cell count can reach 1.4 – 1.6 hundred million/more than ml.
2, prior fermentation temperature adjusting is 33 – 35 DEG C, the time at 12 – 14h, incubation period 8 – 10h.Stop shaking flask in cyclotron oscillation device after terminating, aseptic oxygen supply is cultivated, and inserts biochemical cultivation case.Carry out anaerobic process yeast phase.At this moment yeast cell is healthy and strong, loose, neat.
3, lord ferment period temperature adjusting is 35 – 36 DEG C, time 26 – 30h, is the critical period of producing and ethanol.The inhibition of aerobic yeast cell is there is in acid hydrolysis and enzyme hydrolyzate.And this part inhibition has sub-fraction to be small molecules lipid acid, also produces a certain amount of inhibition in hydrolyzed solution, but its be specifically what and content how much unknown.These inhibitions are all low-molecular-weight materials, less on lord ferment period impact.Prozyme generates ethanol to the hydrolysis of hydrolyzed solution and yeast fermentation to carry out continuously at unified pods.The product of such combinative enzyme hydrolysis---glucose.Because the fermentation of yeast is constantly utilized, eliminate the too high feedback inhibition to prozyme of glucose concn, improve fermenting speed.
4, secondary fermentation phase temperature drops to 31 – 33 DEG C at leisure from 35 – 36 DEG C, and the time is 18 – 20h.Expire (yeast phase 72h) along with fermentation time residual concentration of reduced sugar reduces, and alcoholic strength and ethanol production all reach maximum value, and maturing fermentation wine with dregs supernatant liquor in golden yellow, and obtains satisfied result.
5, vinasse acid hydrolysis and enzyme hydrolyzate maturing fermentation wine with dregs concentration of reduced sugar are at below 0.32g/g, and alcoholic strength is at more than 80.2g/L, and the yield of liquor is 24%.In fermenting-ripening mash, determining concentration of alcohol result is as Fig. 1;
High temperature resistant wine brewing high-activity yeast as seen from the figure, mainly concentrates on 24h-64h fermentation in mid-term and substantially terminates.Now participate in concentration of reduced sugar in fermented liquid and be only 0.32g/g, alcohol concn reaches 80.2g/L.Alcohol transformation efficiency is 240g/kg vinasse.
It can thus be appreciated that vinasse acid hydrolysis liquid adds prozyme after high temperature with process, add high temperature resistant high temperature high-activity yeast of make wine after hydrolysis and enzymolysis and can produce alcohol fuel completely, and often 4.2kg vinasse can produce 1.0kg alcohol fuel.
Claims (1)
1. bent vinasse fermentation produces a method for alcohol fuel, it is characterized in that comprising the following steps:
(1) vinasse 480-500g is got, dry, pulverize, cross 0.8-1.0mm sieve plate hole, be placed in 3L fermentation flask, the warm water adding 1400-1500ml, 32-34 DEG C is sized mixing, and adds urea 13-15g, ammonium sulfate 10-15g, drips sulfuric acid after stirring, regulate PH to 3.5-4.0, with three layers of gauze wrapping sealing;
(2) put into steam sterilizer, at 0.1-0.105MPa, 120-122 DEG C, carry out high-temperature sterilization 110-130 minute, step-down is lowered the temperature, take out, when being cooled to 48-50 DEG C to temperature, the prozyme adding system weight of material 1.8-2.5% be hydrolyzed, liquefy, saccharification process, described prozyme is the Thermostable α-Amylase of 1:1:1:0.5-1 by mass ratio, saccharifying enzyme, cellulase, aspartic protease mixing composition, process and be cooled to 31-33 DEG C after 60-65 minute, add the aspartic protease of above-mentioned equal quality, the dry yeast activation solution brought back to life, the dry yeast activation solution of described resurrection is by yeast temperature 35-40 DEG C, the syrup rehydration 15-20 minute of concentration 2-3%, activation preparation in 1-2 hour when being then cooled to 32-33 DEG C, the inoculum size of the dry yeast activation solution brought back to life is the 12-15% of system weight of material, be placed in the cyclotron oscillation device shaking flask constant temperature culture of 100r/min in sterilisable chamber, 8-10h is cultivated with two-layer felt or three layers of gauze sealing, sampling microscopy yeast count and bud ratio, then the aseptic biochemical cultivation case putting into 32-36 DEG C ferments, by 4h, 8h, 12h respectively vibrates and stirs once, stir once every 7-8h later, middle and later periods fermentation is entered after 40h, stir once every 12h, 72h fermentation ends, take out fermenting-ripening wine with dregs, both alcohol fuel was obtained again by distillation.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698817A (en) * | 2009-10-19 | 2010-04-28 | 李家民 | Method for producing white wine by distilled grain continuous solid state fermentation |
| CN103571878A (en) * | 2012-07-31 | 2014-02-12 | 青岛嘉能节能环保技术有限公司 | Composite agent for producing fuel ethanol by jerusalem artichoke powder fermentation |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698817A (en) * | 2009-10-19 | 2010-04-28 | 李家民 | Method for producing white wine by distilled grain continuous solid state fermentation |
| CN103571878A (en) * | 2012-07-31 | 2014-02-12 | 青岛嘉能节能环保技术有限公司 | Composite agent for producing fuel ethanol by jerusalem artichoke powder fermentation |
Non-Patent Citations (6)
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| 任海伟等: ""白酒丢糟糖化条件的优化及乙醇发酵"", 《应用与环境生物学报》 * |
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