CN101376905B - 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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- CN101376905B CN101376905B CN2008102009908A CN200810200990A CN101376905B CN 101376905 B CN101376905 B CN 101376905B CN 2008102009908 A CN2008102009908 A CN 2008102009908A CN 200810200990 A CN200810200990 A CN 200810200990A CN 101376905 B CN101376905 B CN 101376905B
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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
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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 to producing fermentable sugar by red ramie bark fibre enzymolysis.The fermentable sugar that the inventive method generates can be used for the production of alcohol fuel.
Background technology
At present, human very large to the demand of the energy, but the resources such as oil belong to non-renewable resource, so the various novel energies of people's Devoting Major Efforts To Developing.
Ethanol both can burn separately, can be used as again a kind of good Gasoline oxygenate and high-octane rating blend component, had been acknowledged as one of the most promising renewable and clean energy resource.Its production and be applied in and be in the world high speed development trend, fuel ethanol industrial has become national governments and has solved the petroleum resources shortage problem, 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 strive grain with people and animals, and raw materials cost ratio in total production cost is very high mainly take food crop such as amylaceous corn, wheats as raw material.Be subjected to the restriction of grain resource deficiency, at present, the biomass fuel production take grain as raw material has not possessed expands large-scale resources supplIes again.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 to a great extent agricultural wastes 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, namely finishes the saccharifying of cellulose materials.
Mierocrystalline cellulose is topmost composition in the plant cell wall, is by the linear long-chain macromolecule that 8000～12000 β-D-Glucopyranose monomer is formed by connecting with β-Isosorbide-5-Nitrae-glycosidic link, and 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 technique.Recent two decades comes, microbe fermentation method be the enzymic fermentation production of cellulosic ethanol as a kind of mode of production of environmental protection, obtained countries in the world biologists' favor.The technique that direct bioconversion natural fiber cellulosic material generates ethanol is low because of its cost, equipment simply has good development prospect.
The cellulosic process of cellulose degraded is from it is adsorbed onto on the Mierocrystalline cellulose, 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, Mierocrystalline cellulose thoroughly can be hydrolyzed without any a kind of enzyme.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 form the multienzyme complex structure and work.
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 downcuts continuously and discharges the soluble fiber disaccharides from cellulose molecular chain one end; Beta-glucosidase is hydrolyzed into small molecular sugar with intermediate product-cellobiose and cell-oligosaccharide, thereby removes them to the product inhibition of endoglucanase and exoglucanase.These three hydrolytic processes are carried out simultaneously, with the degradable glucose that becomes of Mierocrystalline cellulose.
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, makes it be difficult to directly be biodegradable.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 that enzymic fermentation is produced 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 processes.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.The method comprises: ramie raw materials is carried out biological degumming, rear ramie bast fiber after coming unstuck is adopted the saccharification of degrading of zytase and acidic cellulase, the recycling saccharomycetes to make fermentation is produced alcohol fuel.In an embodiment, adopt disclosed particular organisms method for degumming in the Chinese patent application 95112564.Yet, the method length consuming time, the industrialization difficulty is large.
US Patent No. 4649113 discloses a kind of method of processing the non-wood material with alkaline peroxide.Yet the method is that sugared conversion coefficient only is about 58% with cellulose conversion.
Therefore, this area easy in the urgent need to developing, utilize Mierocrystalline cellulose to produce the technique 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 so that the pectin of fiber surface and fiber break away from, then pectin is removed in washing, 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 the 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, such as 6.0h).
In another preference, the acid cellulose enzymic activity that step (c) adds is 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 through steam explosion that acquisition is pulverized.
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 the fermentation of producing ethanol.
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 so that the pectin of fiber surface and fiber break away from, then pectin is removed in washing, 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 the 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 of step (c) being produced produces the glycogen material of the fermenting process of ethanol as fermentation, thereby produces ethanol.
Embodiment
The inventor adopts and improves technique 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 degumming process and steam explosion pre-treatment) to bluish dogbane jute skin fiber cellulosic material, then generates fermentable sugar through enzymatic hydrolysis.Pretreated bluish dogbane jute skin fiber cellulosic material about 6 hours, can be the sugar that can be used for fermenting with the cellulose conversion of 85-92% with the acid cellulose enzymic hydrolysis.Compared with the prior art, the present invention has short, transformation efficiency high for the treatment of time.Bastose element extraction yield 90%-95%, cellulose sugar rate are 85%-92%.
Particularly, the inventor adopts physical method and enzyme technology for 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 first the pectin on de-fibering surface by boiling water bath; Then after steam explosion and the pulverization process, thereby so that the connecting key between the fibrous bundle partly interrupts, 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 the production of alcohol fuel.
Compared with the prior art, the present invention has short, transformation efficiency high for the treatment of time.Bluish dogbane jute skin fiber element extraction yield 90%-95%, cellulose sugar rate are 85%-90% (processing 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, diameter 1-3 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 so that the pectin of fiber surface and fiber break away from, 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, such as utilizing the equipment such as pressure kettle.
As used herein, term " aqueous solvent " comprises the mixed solvent that water and water and alcohol (such 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 the materials such as some salt (such 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 usually substantially removing fiber surface gets final product.
The steam explosion step
Only the bluish dogbane jute skin fiber after 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 structure of natural fiber raw material, improves significantly it to the susceptibility of enzyme effect.
Behind the steam explosion, optionally carrying out pulverising step, is 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 obtains, 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 again cellulase system, and a complicated enzyme that is comprised 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) are such as the Rui Shi wood in the Trichoderma mould (T.reesei), koning trichoderma (T.Koningii), Penicillium decumbens (P.decumbens) in 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 (EC3.2.1.4 is called for short EG for Isosorbide-5-Nitrae-D-glucanohydrolase or endo-1,4-β-D-glucanase) is between 23～146KDa.This fermentoid acts on the noncrystalline domain of cellulosic molecule inside, and random hydrolysis β-Isosorbide-5-Nitrae-glycosidic link with the brachymemma of long-chain fiber molecule, produces in a large number the Mierocrystalline cellulose small molecules with non reducing end, such as short chain Mierocrystalline cellulose, cell-oligosaccharide etc.Isomerase EG I such 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 only have 6.3KDa.
(2) circumscribed-β-Isosorbide-5-Nitrae-dextranase, or claim cellobiohydrolase (EC3.2.1.91 is called for short CBH for Isosorbide-5-Nitrae-β-D-glucancellobiohydrolase or exo-1,4-β-D-glucanase), molecular weight is between 38～118KDa.This fermentoid acts on reduction or the non-reduced end of cellulosic molecule, and hydrolysis β-Isosorbide-5-Nitrae-glycosidic link is cut next cellobiose molecule at every turn, thereby destroys cellulosic molecule.Have two kinds such as the mould CBH of wood, CBH I molecular weight is about 66KDa, and CBHII is about 53KDa.
(3) β-1,4-glucuroide (β-Isosorbide-5-Nitrae-glucosidase, EC3.2.1.21, be called for short GE) arranged dividing of interior (or cell wall) enzyme of born of the same parents and extracellular enzyme, 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-Glucose residue and terminal fracture thereof, discharges simultaneously β-D-Glucose and corresponding aglucon, such as aromatic base and hydroxyl etc.In the cellulose hydrolysis process, the beta-glucoside endonuclease capable is hydrolyzed low-molecular-weight cellodextrin and cellobiose, generates glucose, thereby removes the restraining effect to cellulase activity that causes because of the cellobiose accumulation.Therefore, beta-glucosidase is one of the main component of the cellulase system of broad sense, and the height that its enzyme is lived directly affects the action effect of whole cellulase system.
Except above-mentioned three large components, participate in addition some small peptide compounds, " hydrogen bond enzyme ", the cellobiose dehydrogenase etc. 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 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, the operation such as physics comes unstuck, wash fiber crops, steam explosion, pulverizing, acid cellulose Enzymatic hydrolysis.
(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 for subsequent use.
(2) the physics boiling water bath that comes unstuck boils 10-60 minute (such as 30min).The interchangeable water rehydration of laying equal stress on is boiled (such 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 (such as 3MPa, 90s) is dry, for subsequent use.
(5) pulverizing is crushed to the 60-80 order with pulverizer.
(6) fully and completely diastatic fiber is plain for the acid cellulose Enzymatic hydrolysis, the about 500-1000IU/ml of acid cellulose enzymic activity, and pH value 4.8 ± 0.5, enzymic catalytic reaction temperature are 55 ± 2 ℃, reaction times 4-8 hour (such 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 the fermentation industry of producing ethanol.
Utilize various engineering bacterias, the technique by fermentative production of ethanol is well-known.In these techniques, all can use the plain sugar of degrading and producing of bastose of the inventive method preparation.In brief, be exactly the sugar that in zymotechnique, partly or entirely uses the inventive method preparation.
Being compared as follows shown in the table of bastose ethanol and existing stalk fibre ethanol:
| 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 yuan/ton | 5000 yuan/tons or higher |
Can find out that 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 worth 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, and sugared transformation efficiency is high.
(c) the inventive method whole efficiency is high, and 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 for explanation the present invention 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.(purchasing 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 for subsequent use.
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, for subsequent use.
4, pulverize: be crushed to 80 orders with the plant pulverizer.
5, acid cellulose Enzymatic hydrolysis: fully and completely diastatic fiber is plain, and concentration of substrate is 3%, acid cellulose enzymic activity 750IU/ml, and pH value 4.8, enzymic catalytic reaction temperature are 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 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 for subsequent use.
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, for subsequent use.
4, pulverize: be crushed to 80 orders with pulverizer.
5, acid cellulose Enzymatic hydrolysis: fully and completely diastatic fiber is plain, and concentration of substrate is 3%, acid cellulose enzymic activity 700IU/ml, and pH value 4.8, enzymic catalytic reaction temperature are 55 ℃, reaction times 6.5h, thereby the reducing sugar that acquisition is got by cellulose degradation.
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 for subsequent use.
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, for subsequent use.
5, pulverizing is crushed to 60 orders with the plant pulverizer.
6, fully and completely diastatic fiber is plain for the acid cellulose Enzymatic hydrolysis, and concentration of substrate is 3%, acid cellulose enzymic activity 1500IU/ml, and pH value 4.8, enzymic catalytic reaction temperature are 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 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 for subsequent use.
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, for subsequent use.
5, pulverizing is crushed to 90 orders with the plant pulverizer.
6, fully and completely diastatic fiber is plain for the acid cellulose Enzymatic hydrolysis, and concentration of substrate is 3%, acid cellulose enzymic activity 1000IU/ml, and pH value 4.8, enzymic catalytic reaction temperature are 55 ℃, reaction times 5.0h, thereby the reducing sugar that acquisition is got by cellulose degradation.
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 for subsequent use.
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, for subsequent use.
5, pulverizing is crushed to 80 orders with the plant pulverizer.
6, fully and completely diastatic fiber is plain for the acid cellulose Enzymatic hydrolysis, and concentration of substrate is 2.5%, acid cellulose enzymic activity 800IU/ml, and pH value 4.8, enzymic catalytic reaction temperature are 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 bluish dogbane jute skin fiber of the present invention take 3% is as experimental raw, and the cellulose hydrolysis conversion coefficient after the technology of the present invention is implemented to process is 85%-91%.
Embodiment 6
Fermenting experiment
Adopt high-temperature resistant and highly-active dry yeast (purchasing 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, then temperature is down to below 34 ℃ and is activated 1-2h, 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 as each piece document is quoted separately as a reference.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 limited range 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, thus so that the pectin of fiber surface and fiber break away from, then pectin is removed in washing, 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 the 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;
Wherein, the described step of coming unstuck is to boil the bluish dogbane jute skin fiber 90-100 ℃ of water-bath, time 0.5-300 minute; The described step of coming unstuck is carried out in pressure kettle.
2. the method for claim 1 is characterized in that, described usually time is 1-100 minute.
3. the method for claim 1 is characterized in that, described usually time is 2-60 minute.
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 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 through steam explosion that acquisition is pulverized.
9. the purposes of the sugar of the cellulose degradation of method preparation as claimed in 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 sugar of the cellulose degradation of the described method of the claim 1 preparation glycogen material as fermentation.
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| CN103898616B (en) * | 2014-01-23 | 2016-09-07 | 青岛大学 | A kind of sisal hemp degumming technique |
| CN109182418B (en) * | 2018-09-25 | 2020-02-07 | 中国农业科学院麻类研究所 | Method for saccharifying straw through microbial enzymolysis |
| 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 |
| CN114272165B (en) * | 2022-01-11 | 2024-01-16 | 广州亿人康生物医药科技有限公司 | Skin-whitening and tightening plant essence skin-care emulsion and preparation method thereof |
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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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| 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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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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