CN101376905A - Method for producing fermentable sugar by red ramie bark fibre enzymolysis - Google Patents
Method for producing fermentable sugar by red ramie bark fibre enzymolysis Download PDFInfo
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- 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
Abstract
The invention provides a method for producing fermentable sugar through the zymohydrolysis of kenaf bast fiber, in particular the invention provides a method for preparing sugar through the degradation of cellulose, which comprises the following steps: physical degumming, steam blasting and zymohydrolysis are conducted to the kenaf bast fiber, so as to produce the sugar. The fermentable sugar produced by the method can be used for the production of fuel ethanol, and is characterized by short treatment time, high conversion rate, light pollution and the like.
Description
Technical field
The present invention relates to the bioenergy field, more preferably, relate to a kind of novel method producing fermentable sugar by red ramie bark fibre enzymolysis.The fermentable sugar that the inventive method generates can be used for production of fuel ethanol.
Background technology
At present, human very big to the demand of the energy, but resources such as oil belong to non-renewable resource, so people develop various novel energies energetically.
Ethanol both can burn separately, can be used as a kind of good gasoline oxygenation agent and high-octane rating blend component again, had been acknowledged as one of the most promising renewable and clean energy resource.Its production and be applied in and be high speed development trend in the world, fuel ethanol industrial has become national governments and has solved the petroleum resources shortage problem, the important policies means and the economic leverage of protection city atmospheric environment quality and the imbalance between supply and demand of regulation and control agricultural-food.According to estimates, the conservative estimation of China's alcohol fuel demand also will reach 5,000,000 tons every year.
Current ethanol fermentation industry is raw material with food crop such as amylaceous corn, wheats mainly, strive grain with people and animals, and raw materials cost ratio in total production cost is very high.Being subjected to the insufficient restriction of grain resource, at present, is that the biomass fuel production of raw material has not possessed the resources supplIes that enlarges scale more with grain.And the natural fiber cellulosic material is converted into alcohol fuel by biological process, can not only provide a considerable number of, economically viable novel energy for the mankind, and can alleviate agricultural wastes to a great extent to the pollution that ecotope causes, have important economy and ecological significance.
Utilizing the key of natural cellulose material produce alcohol fuel, is to be cellulose hydrolysis fermentable sugars, promptly finishes the saccharifying of cellulose materials.
Mierocrystalline cellulose is a topmost composition in the plant cell wall, be by 8000～12000 β-D-Glucopyranose monomer with β-1, the linear long-chain macromolecule that the 4-glycosidic link is formed by connecting, its glucose subunit is arranged closely.Simultaneously, there is intermolecular misaligned amorphous domain.Mierocrystalline cellulose is usually by hemicellulose, and xylogen surrounds, and forms the mixture of more difficult decomposition.Therefore, Mierocrystalline cellulose is water insoluble, is difficult to hydrolysis.
Hydrocellulose can be taked chemistry or biological method.Biological process is that the enzymic hydrolysis process has the reaction conditions gentleness, and the characteristics of the few or no coupling product of by product are considered to most promising technology.Recent two decades comes, and microbe fermentation method is the mode of production of enzyme process fermentative production cellulosic ethanol as a kind of environmental protection, has obtained countries in the world biologists' favor.Direct bio-transformation natural fiber cellulosic material generation alcoholic acid technology is low because of its cost, equipment simply has bright development prospect.
The cellulosic process of cellulose degraded is adsorbed onto on the Mierocrystalline cellulose from it, and this degradation process can be the effect of single enzyme, as existing one kind of multiple components in conjunction with the cellulase complex body that forms in the Clostridium thermocellum; Also can be that plurality of enzymes is collaborative finishes.In view of the complicacy of cellulosic structure, in fungi, can be without any a kind of enzyme with the thorough hydrolysis of Mierocrystalline cellulose.No matter be in the cellulase complex body, or in the fungus body, cellulosic degraded needs the synergy of plurality of enzymes component.Fungi is by being secreted into the outer free cellulose enzyme of born of the same parents, with lytic enzyme mechanisms of degradation Mierocrystalline cellulose; The bacteria cellulose enzyme then is to work to form the multienzyme complex structure.
At present, the cellulase hydrolysis mechanism of accepting extensively the most is the synergy mechanism of endoglucanase, exoglucanase and beta-glucosidase.This mechanism is to the effect that: the beta-glucan glycosidic bond of endoglucanase random hydrolysis cellulose molecular chain inside forms new chain end; Circumscribed dextran glycosides enzyme is from cellulose molecular chain one end, downcuts continuously and discharges the soluble fiber disaccharides; Beta-glucosidase is hydrolyzed into small molecular sugar with intermediate product-cellobiose and cell-oligosaccharide, thereby removes their product inhibitions to endoglucanase and exoglucanase.These three hydrolytic processes are carried out simultaneously, and Mierocrystalline cellulose is degraded into glucose fully.
The structure of natural fiber cellulosic material is very complicated, mainly is cellulosic highly crystalline and lignifying, has hindered enzyme and has contacted with cellulosic, and it is difficult to directly by biological degradation.At present, concerning most of natural fiber cellulosic material, if do not pass through suitable pre-treatment, directly carry out enzymically hydrolyse, enzymatic hydrolyzation is all very low (＜20%) generally.
The method of enzyme process fermentative production of ethanol has direct fermentation, indirect fermentation method, fermented by mixed bacterium method, simultaneous saccharification and fermentation method, non-isothermal simultaneous saccharification and fermentation method (NSSF), immobilized cell fermentation method etc.Wherein, the simultaneous saccharification and fermentation method because of save time, efficiently, the economic dispatch advantage, become the common method that bio-ethanol transforms.
A kind of method of existing production alcohol fuel is to be the lignocellulosic material production alcohol fuel of representative in order to crop stalk.Yet stalk distributes as agricultural wastes and disperses, and specific volume is very low, causes collection, transportation difficulty, has increased raw materials cost.Usually need 6-7 tons of stalks could produce 1 ton of ethanol, raw materials cost just has a significant impact total cost.
In addition, the content of cellulose of straw is low, and higher degree of lignification has limited the wherein utilization of cellulose components.Publication number is to disclose a kind of method of producing alcohol fuel with the straw kind fibrous material in the CN1880415A Chinese patent application, carries out lignin degradation comprising the method that the straw kind fibrous material is adopted liquid crude enzyme liquid degradation of rice straw and handles.Yet the preparation of this liquid state crude enzyme liquid is comparatively cumbersome, and the degradation effect of xylogen is still difficult satisfactory.
Disclose a kind of enzymic degradation ramie bast fiber in the Chinese patent application 200510124496.4 and produced the method for alcohol fuel.This method comprises: ramie raw materials is carried out biological degumming, and the saccharification of degrading of zytase and acidic cellulase is adopted to the ramie bast fiber after coming unstuck in the back, utilizes saccharomycetes to make fermentation to produce alcohol fuel again.In an embodiment, adopt disclosed particular organisms method for degumming in the Chinese patent application 95112564.Yet, this method length consuming time, the industrialization difficulty is big.
U.S. Pat 4649113 discloses a kind of method of handling the non-wood material with alkaline peroxide.Yet this method is that sugared conversion coefficient only is about 58% with cellulose conversion.
Therefore, this area press for exploitation easy, utilize Mierocrystalline cellulose to produce the technology of fermentable sugars efficiently.
Summary of the invention
Purpose of the present invention just provide a kind of easy, utilize Mierocrystalline cellulose to produce the novel method of fermentable sugars efficiently.
In a first aspect of the present invention, a kind of method for preparing the sugar of cellulose degradation is provided, comprise step:
(a) come unstuck: poach bluish dogbane jute skin fiber in water or aqueous solvent, thus make the pectin of fiber surface and fiber break away from, pectin is removed in washing then, thus the bastose after obtaining to come unstuck;
(b) steam explosion: the bluish dogbane jute skin fiber after coming unstuck is carried out steam explosion, thereby obtain bastose through steam explosion; With
(c) enzymolysis Mierocrystalline cellulose: the bluish dogbane jute skin fiber through steam explosion is carried out enzymolysis, thereby produce sugar.
In another preference, the described step of coming unstuck is to boil the bluish dogbane jute skin fiber 90-100 ℃ of water-bath, and about 0.5-300 of time minute preferably is 1-100 minute, more preferably is 2-60 minute.
In another preference, the described step of coming unstuck is carried out in pressure kettle.
In another preference, the described bluish dogbane jute skin fiber in the step (a) is that length is 0.2-2 centimetre, wide 0.5-2.5 centimetre bluish dogbane fiber crops skin graft section.
In another preference, in step (c) with acid acidic cellulase to the Mierocrystalline cellulose saccharification of degrading.
In another preference, the concentration 2-5wt% of acidic cellulase, pH value 4.0-6.0 (more preferably 4.5-5.5), temperature is 50-55 ℃, the 2-24 hour enzymic catalytic reaction time (preferably 3.0-12 hour is 4-8 hour more preferably, as 6.0h).
In another preference, the acid cellulose enzymic activity that step (c) adds is a 500-1000IU/ml hydrolysis reaction liquid.
In another preference, in step (b), also comprise the bluish dogbane jute skin fiber through steam explosion is carried out pulverization process, thus the bluish dogbane jute skin fiber that acquisition is pulverized through steam explosion.
In another preference, the mean size through the bluish dogbane jute skin fiber of steam explosion of described pulverizing is the 20-120 order, preferably 30-100 order, more preferably 40-80 order.
In a second aspect of the present invention, the purposes of sugar of the cellulose degradation of the method for the invention preparation is provided, they are used as the glycogen material of zymotechnique.
In another preference, described fermentation is to produce the alcoholic acid fermentation.
In a third aspect of the present invention, a kind of method of fermentative production of ethanol is provided, comprise step: in fermentation, with the glycogen material as fermentation of the sugar of the cellulose degradation of the method for the invention preparation.
In a fourth aspect of the present invention, a kind of method of fermentative production of ethanol is provided, comprise step:
(a) come unstuck: poach bluish dogbane jute skin fiber in water or aqueous solvent, thus make the pectin of fiber surface and fiber break away from, pectin is removed in washing then, thus the bastose after obtaining to come unstuck;
(b) steam explosion: the bluish dogbane jute skin fiber after coming unstuck is carried out steam explosion, thereby obtain bastose through steam explosion;
(c) enzymolysis Mierocrystalline cellulose: the bluish dogbane jute skin fiber through steam explosion is carried out enzymolysis, thereby produce sugar; With
(d) sugar that step (c) is produced produces the glycogen material of alcoholic acid fermenting process as fermentation, thereby produces ethanol.
Embodiment
The inventor adopts and improves technology through extensive and deep research, thereby suitable industrialization is provided has utilized Mierocrystalline cellulose high efficiency, low cost ground to produce the novel method of fermentable sugars.The inventive method comprises carries out physics pre-treatment (physics comes unstuck and handles and the steam explosion pre-treatment) to bluish dogbane jute skin fiber cellulosic material, generates fermentable sugar through enzymatic hydrolysis then.Through pretreated bluish dogbane jute skin fiber cellulosic material, about 6 hours, can with the cellulose conversion of 85-92% the sugar that can be used for fermenting with the acid cellulose enzymic hydrolysis.Compared with the prior art, the present invention has characteristics such as treatment time weak point, transformation efficiency height.The plain extraction yield 90%-95% of bastose, saccharification of cellulose rate are 85%-92%.
Particularly, the inventor adopts physical method and enzyme technology at the problem that the cellulosic substrate hydrolysis and saccharification method of producing the alcohol fuel employing exists, and provides a kind of physical method pre-treatment, enzymatic hydrolysis to generate the production method of fermentable sugar.
Preferably, the present invention boils the pectin on de-fibering surface by boiling water bath earlier; Then after steam explosion and the pulverization process, thereby make the connecting key between the fibrous bundle partly interrupt, one-piece construction changes, and is easy to the hydrolytic action of cellulase; The effect of acid cellulose enzymatic hydrolysis can be monose or oligosaccharides with the cellulose degradation saccharification, can be used for production of fuel ethanol.
Compared with the prior art, the present invention has characteristics such as treatment time weak point, transformation efficiency height.The plain extraction yield 90%-95% of bluish dogbane jute skin fiber, saccharification of cellulose rate are 85%-90% (handling about 6 hours under acid cellulose enzymic activity 750IU/ml).
As used herein, " bluish dogbane " bluish dogbane (formal name used at school) has another name called mestha, wild flax, gombo hemp etc., belongs to Dicotyledoneae Malvaceae rose of Sharon plant.
Usually, the high about 1.5-3.5 rice of bluish dogbane cane, the about 1-3 of diameter centimetre.Bluish dogbane is the very strong fast growing and high yield crop of a kind of adaptability, just can gather in the crops in 5-7 month, and output is 16-20t/hm
2
Pre-treatment step:
Pre-treatment of the present invention generally comprises step: physics comes unstuck, washes fiber crops, steam explosion and pulverizes.
In the present invention, it is usually boiling bluish dogbane jute skin fiber in 90-100 ℃ of water or aqueous solvent that physics comes unstuck, thereby makes the pectin of fiber surface and fiber break away from, and about 0.5-300 of poach time minute preferably is 1-100 minute, more preferably is 2-60 minute.Certainly, also can under higher temperature, carry out boiling, for example utilize equipment such as pressure kettle.
As used herein, term " aqueous solvent " comprises the mixed solvent that water and water and alcohol (as methyl alcohol, ethanol) form.For example, the pure content aqueous solution that is lower than 5% (v/v) can be considered aqueous solvent.In addition, in water of the present invention or aqueous solvent, can add materials such as some salt (as sodium-chlor), alkali (sodium hydroxide), acid (example hydrochloric acid) or alcohol.
The consumption of water or aqueous solvent is not particularly limited during poach, getting final product by submergence bluish dogbane fiber crops skin.
For save energy, should reclaim behind the poach hydro-thermal can or the recycling poach after water.
Wash behind the poach, thus the pectin of removal fiber surface.The number of times of washing is not particularly limited, and the pectin of removing fiber surface usually substantially gets final product.
The steam explosion step
Bluish dogbane jute skin fiber after only pretreated the cleaning can directly carry out steam explosion, thereby obtains the bastose cellulosic material through explosion.
Steam explosion can adopt conventional steam explosion equipment.The steam explosion condition can be with reference to the specification sheets of steam explosion equipment or steam explosion condition routinely.Explosion can destroy the composition and the structure of natural fiber raw material, improves its susceptibility to the enzyme effect significantly.
Behind the steam explosion, can randomly carry out pulverising step, be the bluish dogbane jute skin fiber through steam explosion of the described pulverizing of 30-100 purpose thereby obtain mean size.Pulverising step can be used conventional disintegrating apparatus
Enzymolysis step
Through the bluish dogbane jute skin fiber that pre-treatment step of the present invention obtained, the available enzyme solution is produced the sugar that can be used for fermenting.
Can be used for cellulase of the present invention and be not particularly limited, can be the conventional various cellulases that use in this area.
Cellulase (cellulase) claim cellulase system again, and a complicated enzyme of being made up of various ingredients is to be the general name of one group of enzyme of hydrocellulose and derivative thereof.Cellulase production bacterial strain typical and that obtain broad research mainly concentrates on filamentous fungus, wherein the complete and active higher cellulase strain of enzyme system mainly comprises: wood mould (Trichoderma sp.), mould (Penicillium sp.) and basidiomycetes (Basidiomycetes), and as the Penicillium decumbens (P.decumbens) in the Rui Shi wood in the Trichoderma mould (T.reesei), koning trichoderma (T.Koningii), viride (T.viride) and the Penicillium, little purple mould (P.janthinellum) and penicillium funiculosum (P.funiculosum) etc.
Substrate, action site and releasing product according to cellulase are divided three classes cellulase:
(1) molecular weight of endo-beta-1,4-glucanase (1,4-D-glucanohydrolase or endo-1,4-β-D-glucanase, EC 3.2.1.4, abbreviation EG) is between 23～146KDa.This fermentoid acts on the noncrystalline domain of cellulosic molecule inside, random hydrolysis β-1, and the 4-glycosidic link with the brachymemma of long-chain fiber molecule, produces the Mierocrystalline cellulose small molecules of a large amount of band non reducing ends, as short chain Mierocrystalline cellulose, cell-oligosaccharide etc.Isomerase EG I as fungi is 53KDa, and EG III is about 49.8KDa, and has the restriction endonuclease EG molecular weight of two kinds of fiber Acarasiales to have only 6.3KDa.
(2) circumscribed-β-1, the 4-dextranase, or claim cellobiohydrolase (1,4-β-D-glucancellobiohydrolase or exo-1,4-β-D-glucanase, EC 3.2.1.91, abbreviation CBH), molecular weight is between 38～118KDa.This fermentoid acts on the reduction or the non-reduced end of cellulosic molecule, hydrolysis β-1, and the 4-glycosidic link is cut next cellobiose molecule at every turn, thereby destroys cellulosic molecule.Have two kinds as the mould CBH of wood, CBH I molecular weight is about 66KDa, and CBH II is about 53KDa.
(3) β-1,4-glucuroide (EC 3.2.1.21 is called for short GE for β-1,4-glucosidase) has the branch of interior (or cell wall) enzyme of born of the same parents and extracellular enzyme, and molecular weight is between 47～76KDa (extracellular enzyme) or 90～100KDa (intracellular enzyme).Main catalytic hydrolysis contains the compound of β-glucose former times key, makes irreducibility β-D-glucosyl residue and terminal fracture thereof, discharges β-D-glucose and corresponding aglucon simultaneously, as aromatic base and hydroxyl etc.In the cellulose hydrolysis process, low-molecular-weight cellodextrin of beta-glucoside endonuclease capable hydrolysis and cellobiose generate glucose, thereby remove the restraining effect to cellulase activity that causes because of the cellobiose accumulation.Therefore, beta-glucosidase is one of main component of generalized cellulase system, and the height that its enzyme is lived directly influences the action effect of whole cellulase system.
Except above-mentioned three big components, participate in some small peptide compounds, " hydrogen bond enzyme ", the cellobiose desaturase etc. in addition of cellulose degradation process in the fungi.They come the degraded cellulose molecule by oxygenizement or destruction hydrogen bond.
The consumption of enzyme is not particularly limited, and can be 100-5000IU/ml usually, preferably is 500-2000IU/ml.
A kind of particularly preferred enzyme is an acidic cellulase.Its consumption is generally 100-5000IU/ml, preferably is 200-3000IU/ml, more preferably is 500-1500IU/ml.
In a preference of the present invention, described method comprises: get the raw materials ready, operation such as physics comes unstuck, wash fiber crops, steam explosion, pulverizing, acidic cellulase hydrolysis and saccharification.
(1) the bluish dogbane fiber crops skin of getting the raw materials ready is cut into long 0.5-1cm, and the fragment of wide 1-1.5cm is standby.
(2) the physics boiling water bath that comes unstuck boils 10-60 minute (as 30min).The interchangeable water rehydration of laying equal stress on is boiled (as 2-3 time) 1-5 time.
(3) wash the bluish dogbane jute skin fiber that fiber crops have been come unstuck with the tap water flushing, to eliminate the product that comes unstuck that sticks on the fiber.
(4) steam explosion steam explosion condition: be generally 1.5-5Mpa, about 30-300 of time second (as 3MPa, 90s) is dry, standby.
(5) pulverizing is crushed to the 60-80 order with pulverizer.
(6) acidic cellulase hydrolysis and saccharification diastatic fiber element completely, the about 500-1000IU/ml of acid cellulose enzymic activity, pH value 4.8 ± 0.5, enzymic catalytic reaction temperature are 55 ± 2 ℃, reaction times 4-8 hour (as 6.0 hours).
Fermentative production of ethanol
Can be used as the glycogen material of various zymotechniques with the sugar of the cellulose degradation of the inventive method preparation, especially as the glycogen material of producing the alcoholic acid fermentation industry.
Utilize various engineering bacterias, the technology by fermentative production of ethanol is well-known.In these technologies, all can use the plain sugar of degrading and being produced of bastose of the inventive method preparation.In brief, be exactly the sugar that in zymotechnique, partly or entirely uses the inventive method preparation.
Bastose ethanol and existing stalk fibre alcoholic acid are compared as follows shown in the table:
| Bastose ethanol | Stalk fibre ethanol | |
| Raw material | Bastose | Stalk fibre |
| The average yield per mu fiber | 300kg | |
| Be degraded into the fermentable sugars transformation efficiency | 70％-80％ | 20％-35％ |
| The raw material acquisition mode | Can self-built raw material planting base | From peasant household, purchase |
| Raw material processing and transportation | Processing stores on the spot | Big area stock, long-distance transportation |
| Production cost | 3000-3500 unit/ton | 5000 yuan/ton or higher |
As can be seen, major advantage of the present invention is:
(a) grow northerly saltings, desert ground etc. of the kenaf raw material that adopts of the present invention do not utilize the soil, have not only administered ecotope but also important economic value is arranged.Compare with other cellulose raw materials, the content of cellulose 70%-78% of bluish dogbane fiber crops skin, degree of lignification is low, is easy to the hydrolytic action of cellulase.
(b) the inventive method is easy and simple to handle, and the treatment time is short, sugared transformation efficiency height.
(c) the inventive method whole efficiency height, cost is low.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, or the condition of advising according to manufacturer.
Universal method and raw material
Acidic cellulase: by the acidic cellulase of Trichodermareesei, standard enzyme unit of activity (endo cellulase): 20000u/g.(buying) in the outstanding person of Zaozhuang City promise biological enzyme company limited
Cellulosic conversion coefficient=gained saccharic amount/(content of cellulose of substrate dry basis * substrate * 1.1)
The measuring method of reducing sugar: adopt the DNS method.Under the condition of boiling, 3,5-dinitrosalicylic acid (DNS reagent) and reducing sugar generation color reaction, the depth of its color is directly proportional with reducing sugar (with glucose meter) content, by measure its absorbancy at 540nm, can obtain producing the amount of reducing sugar.
Embodiment 1
1, get the raw materials ready: 20kg bluish dogbane fiber crops skin is cut into long 0.5-1cm, and the fragment of wide 1-1.5cm is standby.
2, physics comes unstuck: after boiling water bath boils 30min, wash the bluish dogbane jute skin fiber that has come unstuck with tap water, to eliminate the product that comes unstuck that sticks on the fiber, triplicate.
3, steam explosion: steam explosion condition: 3MPa, 90s are dry, standby.
4, pulverize: be crushed to 80 orders with the plant pulverizer.
5, acidic cellulase hydrolysis and saccharification: diastatic fiber element completely, concentration of substrate is 3%, acid cellulose enzymic activity 750IU/ml, pH value 4.8, enzymic catalytic reaction temperature are 55 ℃, reaction times 6.0h, thereby obtain by cellulose degradation reducing sugar.
After above-mentioned art breading, cellulosic sugared transformation efficiency is 91%.
Embodiment 2
1, get the raw materials ready: 20kg bluish dogbane fiber crops skin is cut into long 0.5-1cm, and the fragment of wide 1-1.5cm is standby.
2, physics comes unstuck: after boiling water bath boils 15min, wash the bluish dogbane jute skin fiber that has come unstuck with tap water, to eliminate the product that comes unstuck that sticks on the fiber, triplicate.
3, steam explosion: steam explosion condition: 2.5MPa, 180s are dry, standby.
4, pulverize: be crushed to 80 orders with pulverizer.
5, acidic cellulase hydrolysis and saccharification: diastatic fiber element completely, concentration of substrate is 3%, acid cellulose enzymic activity 700IU/ml, pH value 4.8, enzymic catalytic reaction temperature are 55 ℃, reaction times 6.5h, thereby obtain by cellulose degradation reducing sugar.
After above-mentioned art breading, cellulosic sugared transformation efficiency is 85%.
Embodiment 3
1, the 20kg bluish dogbane fiber crops skin of getting the raw materials ready is cut into long 0.5-1cm, and the fragment of wide 1-2cm is standby.
2, the physics boiling water bath that comes unstuck boils 45min.
3, wash the bluish dogbane jute skin fiber that fiber crops have been come unstuck with the tap water flushing, to eliminate the product that comes unstuck that sticks on the fiber.
4, steam explosion steam explosion condition: 3MPa, 100s are dry, standby.
5, pulverizing is crushed to 60 orders with the plant pulverizer.
6, acidic cellulase hydrolysis and saccharification diastatic fiber element completely, concentration of substrate is 3%, acid cellulose enzymic activity 1500IU/ml, pH value 4.8, enzymic catalytic reaction temperature are 55 ℃, reaction times 6.0h, thus obtain by cellulose degradation reducing sugar.
After above-mentioned art breading, cellulosic sugared transformation efficiency is 87%.
Embodiment 4
1, the 100kg bluish dogbane fiber crops skin of getting the raw materials ready is cut into long 0.5-1.5cm, and the fragment of wide 1-1.5cm is standby.
2, the physics boiling water bath that comes unstuck boils 15min.
3, wash the bluish dogbane jute skin fiber that fiber crops have been come unstuck with the tap water flushing, to eliminate the product that comes unstuck that sticks on the fiber.
4, steam explosion steam explosion condition: 3.5MPa, 120s are dry, standby.
5, pulverizing is crushed to 90 orders with the plant pulverizer.
6, acidic cellulase hydrolysis and saccharification diastatic fiber element completely, concentration of substrate is 3%, acid cellulose enzymic activity 1000IU/ml, pH value 4.8, enzymic catalytic reaction temperature are 55 ℃, reaction times 5.0h, thus obtain by cellulose degradation reducing sugar.
After above-mentioned art breading, cellulosic sugared transformation efficiency is 86%.
Embodiment 5
1, the 200kg bluish dogbane fiber crops skin of getting the raw materials ready is cut into long 0.5-1cm, and the fragment of wide 1-1.5cm is standby.
2, the physics boiling water bath that comes unstuck boils 10min.
3, wash the bluish dogbane jute skin fiber that fiber crops have been come unstuck with the tap water flushing, to eliminate the product that comes unstuck that sticks on the fiber.
4, steam explosion steam explosion condition: 3MPa, 90s are dry, standby.
5, pulverizing is crushed to 80 orders with the plant pulverizer.
6, acidic cellulase hydrolysis and saccharification diastatic fiber element completely, concentration of substrate is 2.5%, acid cellulose enzymic activity 800IU/ml, pH value 4.8, the enzymic catalytic reaction temperature is 55 ℃, reaction times 6.0h, thereby the reducing sugar that acquisition is got by cellulose degradation.
After above-mentioned art breading, cellulosic sugared transformation efficiency is 85%.
Implementation result
The present invention is an experimental raw with 3% bluish dogbane jute skin fiber, and the cellulose hydrolysis conversion coefficient after the technology of the present invention is implemented to handle is 85%-91%.
Embodiment 6
Fermenting experiment
Adopt high-temperature resistant and highly-active dry yeast (buying) in Hubei Angel Yeast company limited
The Angel Yeast fermention medium adds nutrition and forms: yeast powder 0.5%, peptone 0.3%, (NH
4)
2SO
40.1%, potassium primary phosphate 0.2% is regulated pH5.0 with acetic acid.
Distiller's yeast preparation: 0.5g Angel highly active dry yeast is dissolved in 2% the G/W, shakes up, place on 35-40 ℃, 100rmp shaking table and cultivate 15-20min, temperature is reduced to below 34 ℃ and is activated 1-2h then, can do distiller's yeast and use.
As the glycogen material, adopt conventional fermentation condition with the fermentable sugars for preparing among the embodiment 1, wherein leavening temperature is 35 ℃, yeast consumption %0.5%, fermentation time 48h.
The result: after measured, the alcohol concn in the fermenation raw liquid is 7.44g/L.
All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Claims (10)
1. a method for preparing the sugar of cellulose degradation is characterized in that, comprising:
(a) come unstuck: poach bluish dogbane jute skin fiber in water or aqueous solvent, thus make the pectin of fiber surface and fiber break away from, pectin is removed in washing then, thus the bastose after obtaining to come unstuck;
(b) steam explosion: the bluish dogbane jute skin fiber after coming unstuck is carried out steam explosion, thereby obtain bastose through steam explosion; With
(c) enzymolysis Mierocrystalline cellulose: the bluish dogbane jute skin fiber through steam explosion is carried out enzymolysis, thereby produce sugar.
2. the method for claim 1 is characterized in that, the described step of coming unstuck is to boil the bluish dogbane jute skin fiber 90-100 ℃ of water-bath, and about 0.5-300 of time minute preferably is 1-100 minute, more preferably is 2-60 minute.
3. the method for claim 1 is characterized in that, the described step of coming unstuck is carried out in pressure kettle.
4. the method for claim 1 is characterized in that, the described bluish dogbane jute skin fiber in the step (a) is that length is 0.2-2 centimetre, wide 0.5-2.5 centimetre bluish dogbane fiber crops skin graft section.
5. the method for claim 1 is characterized in that, in step (c) with acid acidic cellulase to the Mierocrystalline cellulose saccharification of degrading.
6. method as claimed in claim 5 is characterized in that, the concentration 2-5wt% of acidic cellulase, and pH value 4.0-6.0, temperature is 50-55 ℃, 2-24 hour enzymic catalytic reaction time.
7. method as claimed in claim 5 is characterized in that, the acid cellulose enzymic activity that step (c) adds is a 500-1000IU/ml hydrolysis reaction liquid.
8. the method for claim 1 is characterized in that, in step (b), also comprise the bluish dogbane jute skin fiber through steam explosion is carried out pulverization process, thus the bluish dogbane jute skin fiber that acquisition is pulverized through steam explosion.
9. the purposes of the sugar of the cellulose degradation of method preparation according to claim 1 is characterized in that, is used as the glycogen material of zymotechnique.
10. the method for a fermentative production of ethanol is characterized in that, comprises step: in fermentation, with the glycogen material as fermentation of the sugar of the cellulose degradation of the described method of claim 1 preparation.
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| CN1966693B (en) * | 2005-11-15 | 2010-11-10 | 中国农业科学院麻类研究所 | Process for preparing fuel ethanol by enzyme method degradation of ramie phloem fiber |
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| CN109182418A (en) * | 2018-09-25 | 2019-01-11 | 中国农业科学院麻类研究所 | A kind of method of microorganism enzymatic saccharification stalk |
| CN109112172B (en) * | 2018-09-25 | 2021-03-23 | 中国农业科学院麻类研究所 | Method for saccharifying straw through microbial enzymolysis |
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Application publication date: 20090304 Assignee: Jiangsu Wei Wei Biomass Technology Co., Ltd. Assignor: Zhongwei Bio-Chemical Co., Ltd., Shanghai Contract record no.: 2016990000191 Denomination of invention: Method for producing fermentable sugar by red ramie bark fibre enzymolysis Granted publication date: 20130123 License type: Exclusive License Record date: 20160509 |
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