CN104745641A - Method for generating ethyl alcohol through natural pH fermentation of lignocellulose - Google Patents
Method for generating ethyl alcohol through natural pH fermentation of lignocellulose Download PDFInfo
- Publication number
- CN104745641A CN104745641A CN201310752385.2A CN201310752385A CN104745641A CN 104745641 A CN104745641 A CN 104745641A CN 201310752385 A CN201310752385 A CN 201310752385A CN 104745641 A CN104745641 A CN 104745641A
- Authority
- CN
- China
- Prior art keywords
- lignocellulose
- natural
- producing
- ethanol
- enlarged culturing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
-
- 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
-
- 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
Landscapes
- Organic Chemistry (AREA)
- 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)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for generatinbg ethyl alcohol through natural pH fermentation of lignocellulose. Specific induction cultivation is carried out on specific strains by virtue of a selected amplification culture medium which take contained C5 and C6 as carbon sources, and the strains have good fermentation activity, natural pH fermentation can be realized, and meanwhile the fermentation performance of the strains is good. Meanwhile, meta-acid of a fermentable system can effectively restrain growth of infectious microbes like lactic acid bacteria, and the concentration of lactic acid, acetic acid and the like generated by the infectious microbe in the fermentation liquid is obviously reduced.
Description
Technical field
The invention belongs to field of microbial fermentation, be specifically related to a kind of method of natural pH fermenting lignocellulose producing and ethanol.
Background technology
Many fossil energies as non-renewable in oil etc. are day by day exhausted, make renewable energy source particularly biofuel receive increasing concern, and bring huge business opportunity and social effect.
Ethanol is clean regeneratable liquors fuel, and many countries have brought into use the gasoline-gasohol that with the addition of certain proportion ethanol, to replace the consumption of gasoline.This New-type fuel can alleviate the wear rate of oil, can reduce automobile exhaust pollution again, has great application and development potentiality.China starts to promote the use of gasohol from calendar year 2001, and current gasohol accounts for about 20% of gasoline-like fuel total flow, and is in the impetus increased year by year.
The main raw material that production ethanol uses both at home and abroad is at present the food crop such as corn.Along with the continuous growth of world population, grain worsening shortages, therefore in the long run, food crop are not the desirable feedstock of producing ethanol.
Biomass energy is a kind of important renewable energy source in future source of energy field.In recent years, countries in the world are all in the correlation technique of Devoting Major Efforts To Developing comprehensive utilization biomass, and achieving certain achievement utilizing biomass to produce in alcohol fuel, biofuel, biological hydrogen, biogas etc., the correlation technique of biomass energy is still the focus of various countries' research and development from now on.China has abundant biomass resource, and according to statistics, the output that stalk is often only by China can reach 700,000,000 tons, has contained very considerable biomass energy, still need further development and utilization among this.
At present, adopt the mode of biomass ferment to produce the correlation technique of ethanol, by the lignocellulose for fermentation in biomass, and then produce the correlation technique of ethanol, defined certain industrialized scale.But the following problem of ubiquity: 1) for making fermentation strain be in optimal pH environment, have comparatively strong active, to improve the output of ethanol, have to add the pH of appropriate alkali to fermented liquid in fermenting process regulate in real time, this not only makes the operation in fermenting process become complicated, and very easily because the error effect fermentation production rate of regulation and control, and under the environment adding alkali, be conducive to the growth of the miscellaneous bacterias such as milk-acid bacteria, easily cause the content of miscellaneous bacteria in fermentation system higher.
Summary of the invention
For this reason, technical problem to be solved by this invention is to provide in a kind of fermenting process without the need to regulating the method for the fermenting lignocellulose producing and ethanol of pH.
The method of natural pH fermenting lignocellulose producing and ethanol of the present invention, comprises the following steps:
1) Ho-Purdue yeast 424A(LNH-ST is got) bacterial strain carries out enlarged culturing;
2) get lignocellulose-containing raw material and carry out enzymolysis, obtain the fermention medium of lignocellulose-containing raw material enzymolysis solution;
3) by the strain inoculation after step 1) enlarged culturing to step 2) in the fermention medium that obtains, control temperature 28-35 DEG C, rotating speed 50-100rpm/min, natural pH, fermentation 24-72h obtains fermented liquid;
4) ethanol in the fermented liquid of separation and Extraction step 3), to obtain final product.
Wherein, described Ho-Purdue yeast 424A(LNH-ST) buy for green science and technology company of the U.S. (Green Tech America, Inc).
In described step 1), the culture condition of the step of described enlarged culturing is specially: the pH value 4-6 controlling enlarged culturing base, spread cultivation temperature 28-35 DEG C, rotating speed 100-200rpm/min.
In described step 1), the enlarged culturing base selected by described enlarged culturing step comprises glucose that weight percentage is 2-10% and weight percentage is the wood sugar of 1-6%.Preferably, in the substratum of described enlarged culturing the content of glucose by weight percentage content be 4-6%, Xylose Content 1-4%.
One or more also comprising in described enlarged culturing base in corn mash, molasses, sweet sorghum stalk juice are carbon source, and one or more in soybean cake powder, corn-dodger powder, corn steep liquor, fish meal, peptone, yeast extract paste are nitrogenous source.
The urea that content is 2-5g/L is also comprised and/or content is the inorganic salt of 0.5-4g/L in described enlarged culturing base.Wherein, described inorganic salt are one or more in Sodium phosphate dibasic, SODIUM PHOSPHATE, MONOBASIC, potassium primary phosphate, dipotassium hydrogen phosphate, Secondary ammonium phosphate.Preferably, in the substratum of described enlarged culturing, containing described inorganic salt potassium primary phosphate 1.5g/L and Secondary ammonium phosphate 1.5g/L.
Further, described enlarged culturing is that inoculum size 5%-20% inoculates enlarged culturing to (0.2-0.3) × 10
9/ ml.
Described step 2) in, the step of described lignocellulose-containing raw material enzymolysis is specially: the enzyme adding 100-200fpu filter paper enzyme activity unit of force in described lignocellulose-containing raw material, insulation mixing 48-96h at 40-60 DEG C, obtain the lignocellulosic material enzymolysis solution that sugared concentration is 8-16%, and regulate the pH to 4-6 of enzymolysis solution.
Described lignocellulose-containing raw material is selected from maize straw, corn cob, hardwood, cork, nutshell, grass, paper, barley-straw, wheat-straw, leaf, cottonseed wadding, newspaper, withy or oat shell.
From structure, lignocellulose mainly comprises Mierocrystalline cellulose, hemicellulose and xylogen.In preferred embodiments, lignocellulose-containing raw material packet containing at least 30wt%, preferably at least 50wt%, more preferably at least 70wt%, the even more preferably lignocellulose of at least 90wt%.It should be understood that in lignocellulose-containing raw material and can also comprise other component, as protein material, starch, sugar, as fermentable sugar and/or not fermentable sugar.Preferably, in the present invention, indication lignocellulose-containing raw material is maize straw.
Described enzyme is selected from cellulase, hemicellulase, amylase, proteolytic enzyme, glucoamylase or lipase.Wherein, cellulase includes but not limited to cellobiohydrolase (cellobiohydrolase I and cellobiohydrolase II) and endo-glucanase and beta-glucosidase enzyme.
The inhibitor that working concentration is 2-20 unit/mL is also comprised in described lignocellulose-containing raw material enzymolysis solution; Described inhibitor is one or more in penicillin, penbritin, Streptomycin sulphate, paraxin, terramycin, tsiklomitsin.
Also containing the urea of 2-5g/L and the inorganic salt of 0.5-4g/L in described lignocellulose-containing raw material enzymolysis solution.
Preferably, in step 2) described in before lignocellulose raw material enzymolysis, the cooking process adopting this area to commonly use to described lignocellulosic material, pickling process, alkaline purification, steam explosion, colloidal mill pulverize in one or more carry out pre-treatment.
In described step 3), by the strain inoculation after step 1) enlarged culturing to step 2) in the fermention medium that obtains, its inoculum size is 5-20%.
In described step 1), before being also included in described enlarged culturing step, described bacterial classification is carried out the step of seed liquor cultivation, seed liquor culturing step condition used is: control ph 4-6, culture temperature 28-35 DEG C, rotating speed 100-200rpm/min.
The seed liquor substratum of described seed liquor culturing step comprises yeast powder 10g/L, peptone 20g/L, glucose 20g/L.Described seed liquor substratum is yeast powder, peptone, glucose, is mixed with and obtains after packing sterilizing.
Further, described seed liquor is cultivated and is: by described Ho-Purdue yeast 424A(LNH-ST) bacterial strain adopts described seed liquor culture medium culturing to (0.1-0.5) × 10
9/ ml.
Technique scheme of the present invention, has the following advantages compared to existing technology:
(1) method of fermentation producing and ethanol of the present invention, by selecting specific bacterial strain, fermentation producing and ethanol is carried out to lignocellulose-containing raw material enzymolysis solution, in whole fermenting process, the activity influence of change to described bacterial strain of the pH of fermented liquid is less, the fermentation carrying out ethanol when not regulating the natural pH of fermented liquid can be realized, and achieve higher ethanol production, avoid and need constantly to add complex operations that alkali brings and the suppression to product during the fermentation; Meanwhile, because in fermenting process, pH is relatively low, inhibit the growth of the miscellaneous bacterias such as milk-acid bacteria, through measuring, in fermented liquid, the concentration of lactic acid, acetic acid etc. that miscellaneous bacteria produces is obviously lower;
(2) method of fermentation producing and ethanol of the present invention, adopt the selected specific inducing culture carrying out specified strain containing C5 and the C6 enlarged culturing base that is carbon source, make described bacterial classification maintain good fermentative activity, not only achieve the fermentation of nature pH, the leavening property of bacterial classification is better simultaneously.
Embodiment
Embodiment 1
The method of the natural pH fermenting lignocellulose producing and ethanol of the present embodiment, comprises the following steps:
1) described Ho-Purdue yeast 424A(LNH-ST is got) bacterial strain carries out seed culture, enlarged culturing; Its cultivating process is specially:
Seed culture: yeast powder 10g/L, peptone 20g/L, glucose 20g/L are mixed with after packing sterilizing and obtain seed liquor substratum, by described Ho-Purdue yeast 424A(LNH-ST) inoculation is to described substratum, and cultivation 20h reaches (0.2-0.3) × 10 to bacterial classification concentration
9/ ml.
Enlarged culturing: get the thalline obtained in described seed culture, be inoculated into containing spreading cultivation in the shaking flask of substratum by inoculum size 5%, the temperature that spreads cultivation 30 DEG C, rotating speed 200rpm, incubation time 16h, first step thalline 5% is seeded to the second stage and spreads cultivation culture medium culturing, adjust ph is 5, the temperature that spreads cultivation 30 DEG C, rotating speed 200rpm, spread cultivation 16h, and enlarged culturing is to (0.2-0.3) × 10
9/ ml.
The substratum of described enlarged culturing is the substratum that spreads cultivation of 2% glucose, 6% wood sugar, containing glucose 2%(mass percent), wood sugar 6%(mass percent), corn steep liquor (dry matter content 40%) 15g/L, KH
2pO
41.5g/L, (NH
4)
2hPO
41.5g/L; Deionized water or softening water configuration solution, adjust pH6.0, packing sterilizing, after cooling, mixing, adds 3g/L urea.
2) in described lignocellulosic material, add the cellulase of 200fpu filter paper enzyme activity unit of force, at 50 DEG C, insulation mixing 72h, obtains the lignocellulosic material enzymolysis solution that sugared concentration is 8%, regulates pH to 5.0; Wherein, described lignocellulosic material is maize straw.
3) get the bacterial strain obtained in described enlarged culturing, be seeded to step 2 with the inoculum size of 10%) in the described lignocellulosic material enzymolysis solution that obtains, temperature remains on 30 DEG C, shaking flask rotating speed 75rpm/min, natural ph, fermentation 72h.
4) separating step 3) fermented liquid in ethanol, to obtain final product.
Embodiment 2
The method of the natural pH fermenting lignocellulose producing and ethanol of the present embodiment, comprises the following steps:
1) described Ho-Purdue yeast 424A(LNH-ST is got) bacterial strain carries out seed culture, enlarged culturing; Its cultivating process is specially:
Seed culture: yeast powder 10g/L, peptone 20g/L, glucose 20g/L are mixed with after packing sterilizing and obtain seed liquor substratum, by described Ho-Purdue yeast 424A(LNH-ST) inoculation to described substratum, 30 DEG C, pH6.0 is cultured to bacterial classification concentration and reaches (0.1-0.2) × 10
9/ ml.
Enlarged culturing: get the thalline obtained in described seed culture, be inoculated into containing spreading cultivation in the shaking flask of substratum by inoculum size 20%, adjust ph is 4, the temperature that spreads cultivation 35 DEG C, and rotating speed 100rpm, incubation time 14h, enlarged culturing is to (0.2-0.3) × 10
9/ ml.
The substratum of described enlarged culturing is the sweet sorghum stalk juice by weight percentage containing glucose 10%, wood sugar 1%, corn-dodger powder 5g/L, potassium primary phosphate 4g/L; Deionized water or softening water configuration solution, packing sterilizing, after cooling, mixing, adds 2g/L urea.
2) in described lignocellulosic material, add the amylase of 100fpu filter paper enzyme activity unit of force, and working concentration is the tsiklomitsin of 2 units/mL, at 40 DEG C, insulation mixing 48h, obtains the lignocellulosic material enzymolysis solution that sugared concentration is 16%, regulates pH to 4.0; Wherein, described lignocellulosic material is that leaf and cottonseed are wadded a quilt with cotton.
3) get the bacterial strain obtained in described enlarged culturing, be seeded to step 2 with the inoculum size of 20%) in the described lignocellulosic material enzymolysis solution that obtains, temperature remains on 28 DEG C, shaking flask rotating speed 50rpm/min, pH value nature, fermentation 24h;
4) separating step 3) fermented liquid in ethanol, to obtain final product;
Embodiment 3
The method of the natural pH fermenting lignocellulose producing and ethanol of the present embodiment, comprises the following steps:
1) described Ho-Purdue yeast 424A(LNH-ST is got) bacterial strain carries out seed culture, enlarged culturing; Its cultivating process is specially:
Seed culture: yeast powder 10g/L, peptone 20g/L, glucose 20g/L are mixed with after packing sterilizing and obtain seed liquor substratum, by described Ho-Purdue yeast 424A(LNH-ST) inoculation extremely described substratum, be cultured to bacterial classification concentration and reach (0.4-0.5) × 10
9/ ml.
Enlarged culturing: get the thalline obtained in described seed culture, is inoculated into containing spreading cultivation in the shaking flask of substratum by inoculum size 10%, and regulate pH to 6, the temperature that spreads cultivation 28 DEG C, rotating speed 150rpm, incubation time 14h, enlarged culturing is to (0.2-0.3) × 10
9/ ml.
The substratum of described enlarged culturing is the molasses culture medium of 6% glucose, 4% wood sugar, containing glucose 6%(mass percent), 4% wood sugar (mass percent), fish meal 10g/L, SODIUM PHOSPHATE, MONOBASIC 0.5g/L; Deionized water or softening water configuration solution, adjust pH7.0, packing sterilizing, after cooling, mixing, adds 5g/L urea.
2) getting oat shell is lignocellulosic material, after alkali soaks, add the proteolytic enzyme of 150fpu filter paper enzyme activity unit of force, working concentration be the Streptomycin sulphate of 20 units/mL, the inorganic salt SODIUM PHOSPHATE, MONOBASIC of the urea of 2g/L and 4g/L, insulation mixing 96h at 60 DEG C, obtain the lignocellulosic material enzymolysis solution that sugared concentration is 12%, regulate pH to 6;
3) get the bacterial strain obtained in described enlarged culturing, be seeded to step 2 with the inoculum size of 5%) in the described lignocellulosic material enzymolysis solution that obtains, temperature remains on 32 DEG C, shaking flask rotating speed 100rpm/min, pH value nature, fermentation 48h;
4) separating step 3) fermented liquid in ethanol, to obtain final product;
Embodiment 4
The method of the natural pH fermenting lignocellulose producing and ethanol of the present embodiment, comprises the following steps:
1) described Ho-Purdue yeast 424A(LNH-ST is got) bacterial strain carries out seed culture, enlarged culturing; Its cultivating process is specially:
Seed culture: yeast powder 10g/L, peptone 20g/L, glucose 20g/L are mixed with after packing sterilizing and obtain seed liquor substratum, by described Ho-Purdue yeast 424A(LNH-ST) inoculation extremely described substratum, be cultured to bacterial classification concentration and reach (0.4-0.5) × 10
9/ ml.
Enlarged culturing: get the thalline obtained in described seed culture, is inoculated into containing spreading cultivation in the shaking flask of substratum by inoculum size 5%, and regulate pH to be 4, the temperature that spreads cultivation 37 DEG C, rotating speed 200rpm, incubation time 14h, enlarged culturing is to (0.2-0.3) × 10
9/ ml.
The substratum of described enlarged culturing is the corn mash of 4% glucose, 2% wood sugar, containing glucose 4%(mass percent), wood sugar 2%(mass percent), soybean cake powder 7g/L, dipotassium hydrogen phosphate 2.3g/L; Deionized water or softening water configuration solution, packing sterilizing, after cooling, mixing, adds 3g/L urea.
2) corn cob is got and hardwood is lignocellulosic material, after boiling 1h, add the cellulase of 120fpu filter paper enzyme activity unit of force, working concentration be the penbritin of 10 units/mL, the primary ammonium phosphate of the urea of 5g/L and 0.5g/L, at 50 DEG C, insulation mixing 72h, obtains lignocellulosic material enzymolysis solution;
3) get the bacterial strain obtained in described enlarged culturing, be seeded to step 2 with the inoculum size of 10%) in the described lignocellulosic material enzymolysis solution that obtains, temperature remains on 35 DEG C, shaking flask rotating speed 100rpm/min, pH value nature, fermentation 72h;
4) separating step 3) fermented liquid in ethanol, to obtain final product.
Embodiment 5
The method of the natural pH fermenting lignocellulose producing and ethanol of the present embodiment, comprises the following steps:
1) described Ho-Purdue yeast 424A(LNH-ST is got) bacterial strain carries out seed culture, enlarged culturing; Its cultivating process is specially:
Seed culture: yeast powder 10g/L, peptone 20g/L, glucose 20g/L are mixed with after packing sterilizing and obtain seed liquor substratum, by described Ho-Purdue yeast 424A(LNH-ST) inoculation is to described substratum, and cultivation 14h spreads cultivation to (0.4-0.5) × 10
9/ ml.
Enlarged culturing: get the thalline obtained in described seed culture, be inoculated into containing spreading cultivation in the shaking flask of substratum by inoculum size 10%, the temperature that spreads cultivation 30 DEG C, rotating speed 200rpm, incubation time 14h, enlarged culturing is to (0.2-0.3) × 10
9/ ml.
The substratum of described enlarged culturing is the mixture of molasses and sweet sorghum stalk juice, containing glucose 4%(mass percent), wood sugar 5%(mass percent), mixture 5g/L, Sodium phosphate dibasic 1.5g/L, the Secondary ammonium phosphate 1.5g/L of peptone and yeast extract paste; Deionized water or softening water configuration solution, packing sterilizing, after cooling, mixing, adds 3g/L urea.
2) in described lignocellulosic material, add glucoamylase, the urea of 3g/L and the primary ammonium phosphate of 2.3g/L, at 50 DEG C, insulation mixing 72h, obtains lignocellulosic material enzymolysis solution;
3) get the bacterial strain obtained in described enlarged culturing, be seeded to step 2 with the inoculum size of 10%) in the described lignocellulosic material enzymolysis solution that obtains, temperature remains on 30 DEG C, shaking flask rotating speed 80rpm/min, pH value nature, fermentation 72h;
4) separating step 3) fermented liquid in ethanol, to obtain final product.
Comparative example
The method of the production ethanol of this comparative example, get commercially available candida tropicalis Candidatropicalis, the method identical with embodiment 1 is adopted to carry out seed culture and enlarged culturing, and adopt identical fermented liquid, fermentation condition ferments, distinguishing characteristics is only: add ammoniacal liquor in fermenting process and regulate pH, according to on-line pH value varitrol, supplement 20% ammoniacal liquor according to pH value change in real time, pH value is remained between 4.6-5.5.
Effect experimental examples
According to the method ethanol production in embodiment 1-5 and comparative example.
Measure the initial sugared concentration in the fermented liquid after fermentation, terminal sugar concentration, wood sugar rate of consumption, theoretical total reducing sugar alcohol conversion, lactic acid concn and acetic acid concentration.
Its test result is as follows:
It can thus be appreciated that, adopt method of the present invention to produce ethanol, higher alcohol yied can be reached, simultaneously in fermenting alcohol process without the need to carrying out pH regulator, reduce cost, and simplify the operation in fermenting process.In addition, adopt method of the present invention to produce ethanol, the concentration of lactic acid and acetic acid all significantly declines, and illustrates that the content of the miscellaneous bacterias such as milk-acid bacteria in fermentation system is minimized.
Obviously, above-described embodiment is only for clearly example being described, and the restriction not to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And thus the apparent change of extending out or variation be still among the protection domain of the invention.
Claims (11)
1. a method for natural pH fermenting lignocellulose producing and ethanol, is characterized in that, comprise the following steps:
1) Ho-Purdue yeast 424A(LNH-ST is got) bacterial strain carries out enlarged culturing;
2) get lignocellulose-containing raw material and carry out enzymolysis, obtain the fermention medium of lignocellulose-containing raw material enzymolysis solution;
3) by the strain inoculation after step 1) enlarged culturing to step 2) in the fermention medium that obtains, control temperature 28-35 DEG C, rotating speed 50-100rpm/min, natural pH, fermentation 24-72h obtains fermented liquid;
4) ethanol in the fermented liquid of separation and Extraction step 3), to obtain final product.
2. the method for natural pH fermenting lignocellulose producing and ethanol according to claim 1, is characterized in that:
In described step 1), the culture condition of the step of described enlarged culturing is specially: the pH value 4-6 controlling enlarged culturing base, spread cultivation temperature 28-35 DEG C, rotating speed 100-200rpm/min.
3. the method for natural pH fermenting lignocellulose producing and ethanol according to claim 1 and 2, is characterized in that:
In described step 1), the enlarged culturing base selected by described enlarged culturing step comprises glucose that weight percentage is 2-10% and weight percentage is the wood sugar of 1-6%.
4. the method for natural pH fermenting lignocellulose producing and ethanol according to claim 3, is characterized in that:
Also comprise one or more in corn mash, molasses, sweet sorghum stalk juice in described enlarged culturing base for carbon source, and soybean cake powder, corn-dodger powder, corn steep liquor, fish meal, peptone, one or more in yeast extract paste are nitrogenous source.
5. the method for the natural pH fermenting lignocellulose producing and ethanol according to claim 3 or 4, is characterized in that: also comprise the urea that content is 2-5g/L in described enlarged culturing base and/or content is the inorganic salt of 0.5-4g/L.
6. according to the method for the arbitrary described natural pH fermenting lignocellulose producing and ethanol of claim 1-5, it is characterized in that: described step 2) in, the step of described lignocellulose-containing raw material enzymolysis is specially: the enzyme adding 100-200fpu filter paper enzyme activity unit of force in described lignocellulose-containing raw material, insulation mixing 48-96h at 40-60 DEG C, obtain the lignocellulosic material enzymolysis solution that sugared concentration is 8-16%, and regulate the pH to 4-6 of enzymolysis solution.
7. the method for natural pH fermenting lignocellulose producing and ethanol according to claim 6, is characterized in that:
Described lignocellulose-containing raw material is selected from maize straw, corn cob, hardwood, cork, nutshell, grass, paper, barley-straw, wheat-straw, leaf, cottonseed wadding, newspaper, withy or oat shell.
8. the method for the natural pH fermenting lignocellulose producing and ethanol according to claim 6 or 7, is characterized in that:
Described enzyme is selected from cellulase, hemicellulase, amylase, proteolytic enzyme, glucoamylase or lipase.
9. the method for the natural pH fermenting lignocellulose producing and ethanol according to claim 6 or 7 or 8, is characterized in that:
The inhibitor that working concentration is 2-20 unit/mL is also comprised in described lignocellulose-containing raw material enzymolysis solution; Described inhibitor is one or more in penicillin, penbritin, Streptomycin sulphate, paraxin, terramycin, tsiklomitsin.
10., according to the method for the arbitrary described natural pH fermenting lignocellulose producing and ethanol of claim 1-9, it is characterized in that:
In described step 1), before being also included in described enlarged culturing step, described bacterial classification is carried out the step of seed liquor cultivation, seed liquor culturing step condition used is: control ph 4-6, culture temperature 28-35 DEG C, rotating speed 100-200rpm/min.
The method of 11. natural pH fermenting lignocellulose producing and ethanols according to claim 10, is characterized in that: the seed liquor substratum of described seed liquor culturing step comprises yeast powder 10g/L, peptone 20g/L, glucose 20g/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310752385.2A CN104745641B (en) | 2013-12-31 | 2013-12-31 | Method for producing ethanol by fermenting lignocellulose through natural pH |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310752385.2A CN104745641B (en) | 2013-12-31 | 2013-12-31 | Method for producing ethanol by fermenting lignocellulose through natural pH |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104745641A true CN104745641A (en) | 2015-07-01 |
| CN104745641B CN104745641B (en) | 2020-05-12 |
Family
ID=53585897
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310752385.2A Active CN104745641B (en) | 2013-12-31 | 2013-12-31 | Method for producing ethanol by fermenting lignocellulose through natural pH |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104745641B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101289677A (en) * | 2007-04-17 | 2008-10-22 | 中粮集团有限公司 | Process for preparing ethanol by using cellulose-containing raw material |
| CN101899478A (en) * | 2010-07-26 | 2010-12-01 | 安徽丰原发酵技术工程研究有限公司 | Method for producing ethanol by simultaneous high-efficiency saccharification half enzymolysis and fermentation of straw lignocellulose |
| CN102083992A (en) * | 2008-05-07 | 2011-06-01 | 诺维信公司 | Fermentation of a lignocellulose-containing material |
| CN102168113A (en) * | 2011-02-25 | 2011-08-31 | 安徽丰原发酵技术工程研究有限公司 | Method for producing ethanol by using straw lignocellulose raw materials |
-
2013
- 2013-12-31 CN CN201310752385.2A patent/CN104745641B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101289677A (en) * | 2007-04-17 | 2008-10-22 | 中粮集团有限公司 | Process for preparing ethanol by using cellulose-containing raw material |
| CN102083992A (en) * | 2008-05-07 | 2011-06-01 | 诺维信公司 | Fermentation of a lignocellulose-containing material |
| CN101899478A (en) * | 2010-07-26 | 2010-12-01 | 安徽丰原发酵技术工程研究有限公司 | Method for producing ethanol by simultaneous high-efficiency saccharification half enzymolysis and fermentation of straw lignocellulose |
| CN102168113A (en) * | 2011-02-25 | 2011-08-31 | 安徽丰原发酵技术工程研究有限公司 | Method for producing ethanol by using straw lignocellulose raw materials |
Non-Patent Citations (1)
| Title |
|---|
| MING W. LAU等: "Cellulosic ethanol production from AFEX-treated corn stover using Saccharomyces cerevisiae 424A(LNH-ST)", 《PNAS》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104745641B (en) | 2020-05-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101434913B (en) | Wine brewing yeast strain and method for producing ethanol by efficient stalk fermentation | |
| Zhu et al. | A comparison of the production of ethanol between simultaneous saccharification and fermentation and separate hydrolysis and fermentation using unpretreated cassava pulp and enzyme cocktail | |
| CN104774877B (en) | A kind of method of lignocellulose biomass co-producing ethanol, acetone and butanol | |
| Li et al. | A consolidated bio-processing of ethanol from cassava pulp accompanied by hydrogen production | |
| CN101182551B (en) | Method for producing fuel alcohol through alkaline method preprocessing plant fibre | |
| CN102199554B (en) | Saccharomyces cerevisiae strain with multiple-stress resistance, and application thereof in cellulose alcohol fermentation | |
| CN105200094B (en) | A method of utilizing microbial fermentation lignocellulosic material producing and ethanol | |
| CN102174433B (en) | Clostridium beijerinckii with high stress resistance and application thereof | |
| CN101555494A (en) | Method for preparing fuel of ethanol from immobilized mixed strain fermented cellulosic hydrolysate | |
| CN101638673A (en) | Method for manufacturing alcohol by utilizing fermentation of plant straws | |
| CN103898167A (en) | A method of producing ethanol | |
| CN103898166A (en) | Method of producing ethanol | |
| CN104805133B (en) | A method of ethyl alcohol is produced using microorganism common fermentation C5 and C6 | |
| Rai et al. | OPTIMIZATION OF PROCESS PARAMETERS FOR CELLULASE PRODUCTION BY NOVEL THERMOTOLERANT YEAST. | |
| CN104004794A (en) | Method for preparing n-butyl alcohol in fermentation mode by hydrolyzing lignocellulose through mixed cellulose crude enzyme | |
| JP5352882B2 (en) | A method for producing a cellulolytic enzyme using a Clostridium microorganism and a method for culturing and growing the Clostridium microorganism. | |
| CN104263771B (en) | A kind of method using not detoxification cellulosic hydrolysate producing microbial grease | |
| CN104745499A (en) | Amplification culture method for producing ethanol microorganisms by virtue of co-fermentation of C5 and C6 | |
| CN102703523B (en) | Method for producing butanol by mixed fermentation of bagasse and molasses serving as raw materials | |
| CN103103221B (en) | Method for converting cellulose into ethanol by use of mixed culture of genetically recombinant yeast | |
| CN102311977A (en) | Method for producing ethanol by using cassava residues | |
| CN101603056B (en) | Method for fermenting ethanol by utilizing cellulase from animals and microorganisms for cooperating enzymolysis | |
| JP2014176351A (en) | Method for producing ethanol | |
| CN102851325A (en) | Fermentation method for producing ethanol by using enzymatic saccharification of corn cob | |
| CN104745641A (en) | Method for generating ethyl alcohol through natural pH fermentation of lignocellulose |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |