CN101525635B - Method for preparing ethanol by using raw material containing cellulose - Google Patents

Method for preparing ethanol by using raw material containing cellulose Download PDF

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CN101525635B
CN101525635B CN2008101013145A CN200810101314A CN101525635B CN 101525635 B CN101525635 B CN 101525635B CN 2008101013145 A CN2008101013145 A CN 2008101013145A CN 200810101314 A CN200810101314 A CN 200810101314A CN 101525635 B CN101525635 B CN 101525635B
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steam explosion
cellulose
explosion products
weight
raw material
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CN101525635A (en
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岳国君
刘甲申
李春玲
王国忠
林海龙
于天杨
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Cofco Corp
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Cofco Corp
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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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    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Abstract

The invention discloses a method for preparing ethanol by using a raw material containing cellulose, which comprises the steps of subjecting the raw material containing cellulose to steam explosion, mixing an obtained product of the steam explosion with an enzyme for enzymolysis, and fermenting the obtained enzymatic product, wherein a method for the steam explosion of the raw material containing cellulose comprises the steps of continuously delivering the raw material containing cellulose to a heat-preservation pressure unit, keeping the raw material containing cellulose in the heat-preservation pressure unit for 3 to 10 minutes and then releasing part of the raw material containing cellulose in the heat-preservation pressure unit to normal pressure every other 5 to 12 seconds; and a method for mixing the product of the steam explosion and the enzyme comprises the step of mixing the product of the steam explosion with the enzyme in batches, wherein the batches are at least two batches, and in each batch, the additive amount of the product of the steam explosion accounts for 60 percent of the total weight of the product of explosion at most. The method of the invention can effectively improve the sugar conversion rate of the raw material, thereby greatly improving ethanol yield.

Description

Adopt the feedstock production alcoholic acid method of cellulose
Technical field
The invention relates to the alcoholic acid preparation method, more specifically is about adopting the feedstock production alcoholic acid method of cellulose.
Background technology
The raw material by cellulose of prior art prepares the alcoholic acid method like stalk and mainly comprises three steps: 1, the pre-treatment of the raw material of cellulose, 2, cellulosic enzymolysis; 3, the fermentation system ethanol of sugar.
The raw material of cellulose is generally root, stem and the leaf of plant, comprises stalk.Said stalk is root, stem and the leaf that stays behind farm crop (being generally herbage) the results fruit, is mainly stem and leaf, like the stalk of cereal grasss such as corn straw, broomcorn straw, wheat straw.The staple of stalk is a lignocellulose.Said lignocellulose is the mixture of Mierocrystalline cellulose, semicellulose and xylogen, and Mierocrystalline cellulose, semicellulose and xylogen are combined closely through covalent linkage or non covalent bond, and wood fibre accounts for the 70-90% of stalk gross weight.Mierocrystalline cellulose, semicellulose and xylogen content separately can the difference to some extent with the different piece of the farm crop of different sources or farm crop in the stalk.
The pretreatment process of the raw material of cellulose is generally steam explosion and/or acid hydrolysis, to destroy the embedding of xylogen to Mierocrystalline cellulose and semicellulose.Though acid hydrolysis can destroy xylogen, also can destroy Mierocrystalline cellulose, reduce the productive rate of glucose then, and also need add after the acid hydrolysis in a large amount of alkali and could carry out enzymolysis and fermentation.Therefore more tendency is only accomplished preprocessing process through steam explosion, reaches the embedding effect of xylogen to Mierocrystalline cellulose and semicellulose that destroy.
Mierocrystalline cellulose is commonly referred to enzyme digestion reaction by the reaction of cellulase hydrolysis, can be called saccharification react again, and the primary product of enzymolysis is a monose.Mierocrystalline cellulose can change into glucose under the effect of cellulase; Semicellulose can change into monose such as wood sugar under the effect of hemicellulase; Glucose and wood sugar may be used to fermentative production of ethanol; Can prepare ethanol by glucose fermentation such as yeast saccharomyces cerevisiae, pichia stipitis can prepare ethanol by xylose-fermenting.Cellulosic non-crystalline structure is easy to broken, and it can be degraded into glucose fully, becomes ethanol by glucose fermentation then.But cellulosic crystalline texture is difficult to be destroyed.In addition, xylogen is a kind of non-polysaccharide material that is polymerized by phenyl-propane, by the verivate of aromatic hydrocarbon with-C-C-key ,-the O-key is crosslinked together in length and breadth; Its side chain again with semicellulose with covalent bonds, form a very fine and close network structure, Mierocrystalline cellulose tightly is wrapped in the inside; Be unfavorable for that cellulase is to cellulosic attack; Make cellulase can't contact substrate, therefore, need earlier stalk to be carried out pre-treatment with hemicellulase; Contact with cellulase and/or hemicellulase again after steam explosion or the s.t. as stalk is carried out, Mierocrystalline cellulose is hydrolyzed.
CN1117835C discloses a kind of method of dissolve with ethanol steam burst straw lignin prepare liquid fuel, and this method is a raw material with the steam puffed stalk, in autoclave, with the xylogen in the alcohol extraction steam puffed stalk; Obtain filtrating with vacuum filtration again, and steam puffed stalk is extracted slag carry out the synchronous saccharification solid state fermentation of ethanol through, alcohol extraction liquid and vacuum filtration gained filtrating are the liquid fuel of this method preparation.
The preparation method of said steam puffed stalk raw material comprises plant materials such as wheat straw, straw or the cornstalk steam-explosion jar of packing into of chopping, under 1.2-1.7 MPa pressure, keeps pressure 5-10 minute, and afterwards, instant decompression discharges.
The method of the xylogen in the described alcohol extraction steam puffed stalk is in autoclave, adds steam puffed stalk and ethanol, and steam puffed stalk and alcoholic acid volume ratio are 1: 10-1: 50, in the pressure range of 1.5-2.0 MPa, kept 1-2 hour.
Saidly steam puffed stalk is extracted slag carry out the synchronous saccharification solid state fermentation of ethanol through and be meant: in 33-35 ℃ of scope (fermentation), kept 14-16 hour, in 38-40 ℃ of scope (enzymolysis), keep 68 hours, and fermentation 2-3 days that hockets.
CN1193099C discloses a kind of use heat resistant type yeast and from the lignin fiber cellulosic biomass, has produced the alcoholic acid method, and this method may further comprise the steps:
It is the 15-30 millimeter that the lignin fiber cellulosic biomass is ground into particle size; Lignin fiber cellulosic biomass to pulverizing carries out the vapor explosion pre-treatment, keeps the pressure of 1-3 MPa and 190-230 ℃ temperature 1-10 minute time according to the type of material therefor, causes explosive decompression afterwards; Collect pretreated material and pass through filtering separation liquid and solid part, solid part is introduced in the fermentation storage; Adding concentration to the fermentation storage is that 15FPU/ restrains the cellulosic beta-glucosidase of cellulosic cellulase and 12.6 ius/restrain; Give fermentation storage inoculation Marx Crewe Vickers yeast CECT the suspension-s of 10875 heat resistant type yeast cultures; Mixture was shaken under 42 ℃ 72 hours.
Document " to the introduction of straw biomass to alcohol conversion process ", Zhang Xuefeng, orchid are kindly helped secure the success of, wine brewing, the 29th the 4th phase of volume; The 37-38 page or leaf discloses a kind of method of utilizing straw to produce alcohol in July, 2002, and this method comprises to be removed corn straw after the stone deironing cleans; Cut into 1.5 centimetres segment, water logging 40 minutes feeds 2.5 MPa steam, is incubated 8 minutes; Pressure release, the steam explosion products of washing 1/10th, and this wooden enzyme produces cellulase in said 1/10th steam explosion products, cultivating, and the gained culture is mixed with other 9/10ths steam explosion products; Be incubated 50 ℃ of hydrolysis 24 hours, through filtering 6% liquid glucose, mineral membrane is condensed into the liquid glucose more than 20%, adds Xiu Hata yeast (0.8-1.2 hundred million/milliliter); Fermented 24 hours, and produced wine part 10%, distill 99.5% alcohol.
Adopt aforesaid method all can realize feedstock production alcoholic acid purpose, but ethanol yield is lower, generally is no more than 20% by cellulose.
Summary of the invention
The objective of the invention is to overcome the lower defective of feedstock production alcoholic acid method alcohol yied of existing employing cellulose, the feedstock production alcoholic acid method of the higher employing cellulose of a kind of alcohol yied is provided.
Contriver of the present invention finds; In the feedstock production alcoholic acid method of the employing cellulose of prior art, the pretreatment process of the raw material of said cellulose adopts the intermittent type steam explosion usually, and this method comprises that elder generation is delivered to steam blasting device with a collection of cellulose containing raw material; Feed the pressure of steam to steam explosion; And insulation for some time, moment pressure release is then accomplished steam explosion and is collected steam explosion products; And then the raw material of a collection of cellulose is delivered in the steam blasting device, feed the pressure of steam to steam explosion once more, and insulation for some time, moment pressure release is then accomplished steam explosion and is also collected steam explosion products; And according to treatment capacity, the raw material to subsequently cellulose carries out the intermittent type steam explosion according to the method described above.Contriver of the present invention finds, and is all identical like pressure and temperature even guarantee the condition of each steam explosion when adopting such steam explosion method, and still, the explosion degree of the steam explosion products that but can't guarantee to obtain behind each steam explosion is all identical.The reason of inferring possibly be; Because the amount of every batch of raw material of inserting the cellulose in the steam blasting device is more; And the factors such as instability of the difference of the accumulation mode of every batch of raw material or steam explosion condition cause can't guarantee to make and fully absorb water vapor in the short several minutes of whole cellulose containing raw materials after feeding steam; Therefore; Still the raw material that has most of cellulose be owing to can't fully absorb water vapor, and can not be popped by its inner water vapour fully, causes the xylogen sheath of embedding Mierocrystalline cellulose and semicellulose in the raw material of cellulose can't be by abundant destruction; Reduce the cellulose ratios that can contact with enzyme in the raw material of cellulose, thereby reduced ethanol yield.In addition, the method for intermittent type steam explosion consumed time in the pre-treatment of raw material is longer, and the treatment capacity of raw material is lower in the unit time, thereby is unfavorable for the raising of production efficiency.
In addition, in the method for the feedstock production alcohol fuel of the employing cellulose of prior art, the raw material of cellulose; Normally exist like stalk with the form of liquid slurry; When the whole stalks with the slurries shape directly mix with enzyme, enzyme concn is reduced, enzyme activity is descended.Moreover; To pass through pretreated stalk and the method for enzyme mixed enzymolysis is generally in the mixed solution that stalk particle with fragmentation once all joins enzyme and water and mixes with enzyme liquid; Carry out the enzymatic saccharification reaction, perhaps, earlier used reaction mass (through pretreated stalk particle) is mixed with water in retort; Then under hydrolysis temperature, enzyme directly joined in the retort react.Although above-mentioned mixing is carried out under agitation condition, stalk particle still is not easy to mix with enzyme.Because; Diminish through pretreated stalk granularity, particle and clastic stalk particle are easy to mutual adhesion and reunion, therefore once whole stalks are mixed with enzyme fully; Only there is the part stalk fully to contact with enzyme; Because the adhesion between the stalk has the part stalk with reunion and can not fully contact with enzyme, even under agitation, can not fully stalk be broken up.Therefore, behind the enzymolysis cellulose conversion be sugar transformation efficiency low, the ethanol yield that also can cause sugar-fermenting to obtain is lower.
The invention provides a kind of feedstock production alcoholic acid method that adopts cellulose, this method comprises the raw material of steam blasting cellulose-containing; With the steam explosion products that obtains mix with enzyme, enzymolysis; The enzymolysis product that fermentation obtains; Wherein, The method of the raw material of said steam blasting cellulose-containing comprises that the raw material with cellulose is delivered in the heat preservation pressure device continuously; The raw material of cellulose was kept in this heat preservation pressure device after 3-10 minute, and every separated 5-12 makes the raw material of the part cellulose in the said heat preservation pressure device be discharged into normal pressure second; Mix steam explosion products said steam explosion products is comprised with the enzyme method of mixing with enzyme in batches, said number of times in batches is at least 2 times, and the amount of the steam explosion products of every batch of adding is at most 60% of whole steam explosion products weight.
On the one hand; The method that the present invention adopts continuous steam explosion is carried out pre-treatment to the raw material of cellulose; Can make said raw material successive of inserting the whole cellulose in the steam blasting device under stable steam explosion condition, in the heat preservation pressure device, fully absorb aqueous vapor, and make the inner aqueous vapor of raw material of cellulose change into water vapor; Water vapour is gone out by the raw material inside of cellulose when pressure release; Fully the xylogen sheath of embedding Mierocrystalline cellulose and semicellulose in the raw material of destruction cellulose has increased the cellulose ratios that can contact with enzyme in the raw material of cellulose, thereby has improved ethanol yield effectively.And; The method of the raw material of steam blasting cellulose-containing of the present invention is that successive makes the raw material of cellulose in the heat preservation pressure device, be incubated and keep pressure; Successive is accomplished the steam explosion of raw material then; Therefore, the treatment capacity of the raw material of the cellulose that method of the present invention was handled in the unit time is more, helps the raising of production efficiency.On the other hand; Method of the present invention is through mix steam explosion products in batches with enzyme; Make the steam explosion products of every batch of adding mix and fully to contact, and in reaction process, the concentration of enzyme remain on higher level always with enzyme; Therefore, steam explosion products can fully be reacted with enzyme.Thereby improved the sugared transformation efficiency of raw material effectively, therefore improved ethanol yield greatly.
For example, under other enzymatic hydrolysis condition and all identical situation of steam explosion condition, the alcohol yied of embodiment 1 reaches 23.7%, and the alcohol yied of Comparative Examples 1 is merely 18.3%, and the alcohol yied of Comparative Examples 2 is merely 18.6%, and increase rate is up to 29.5-27.4%.And the continuous steam explosion method of embodiment 1 per hour can be handled the raw material 400-500 kilogram of cellulose, and the steam explosion method of Comparative Examples 1 per hour can only be handled the raw material 180-260 kilogram of cellulose.
Description of drawings
Fig. 1 is the electron scanning micrograph of natural corn stalk;
Fig. 2 is the electron scanning micrograph according to stalk behind the method steam explosion of Comparative Examples 1;
Fig. 3 is the electron scanning micrograph according to stalk behind the method steam explosion of Comparative Examples 2;
Fig. 4 is the electron scanning micrograph according to stalk behind the method steam explosion of the embodiment of the invention 1.
Embodiment
According to the present invention, this method comprises the raw material of steam blasting cellulose-containing; With the steam explosion products that obtains mix with enzyme, enzymolysis; The enzymolysis product that fermentation obtains; Wherein, The method of the raw material of said steam blasting cellulose-containing comprises that the raw material with cellulose is delivered in the heat preservation pressure device continuously, and the raw material of cellulose was kept in this heat preservation pressure device after 3-10 minute, be preferably 4-7 minute after; Every at a distance from 5-12 second, be preferably 7-10 and make the raw material of the part cellulose in the said heat preservation pressure device be discharged into normal pressure second; Mix steam explosion products said steam explosion products is comprised with the enzyme method of mixing with enzyme in batches, said number of times in batches is at least 2 times, and the amount of the steam explosion products of every batch of adding is at most 60% of whole steam explosion products weight.
According to the present invention, the raw material of said cellulose is delivered in the heat preservation pressure device continuously, in this device, is transmitted, and in this heat preservation pressure device, is incubated and keeps pressure.The specification of the heat preservation pressure device that the different production scale is adopted is also different; Therefore; The transfer rate of the heat preservation pressure device of different size is also different; The present invention does not limit the transfer limes speed of the raw material of said cellulose is special, and the transfer limes speed of the raw material of said cellulose is as long as guaranteeing the raw material of said cellulose kept 3-10 minute in this heat preservation pressure device, so that the aqueous vapor of formation can fully get in the raw material of cellulose under the temperature of this device, pressure; Water vapour is gone out when the pressure release by the raw material inside of cellulose, with the xylogen sheath of embedding Mierocrystalline cellulose and semicellulose in the raw material of abundant destruction cellulose.
Said heat preservation pressure device can be various heat preservation pressure devices, as long as can make the raw material of the cellulose of continuous conveying in this device, reach the condition of steam explosion.The condition of said continuous steam explosion can adopt the conventional steam explosion condition of prior art, and like the temperature of steam explosion, promptly the temperature of heat preservation pressure device can be 180-200 ℃, more preferably 185-195 ℃; The pressure of said steam explosion, promptly the pressure of heat preservation pressure device can be 1.4-2.0 MPa, more preferably 1.5-1.8 MPa.Under above-mentioned steam explosion condition, can both fully destroy the xylogen sheath of stalk embedding Mierocrystalline cellulose and semicellulose, realize the object of the invention.
Said raw material with the part cellulose in the heat preservation pressure device is discharged into the method that non-pressurized method can adopt various routines, for example, can realize through the pressure valve that control is arranged on the heat preservation pressure device; Said pressure valve is arranged on the exit of heat preservation pressure device, when the pressure in the heat preservation pressure device reaches the pressure of said steam explosion, and valve opening; Make the raw material of the part cellulose in the heat preservation pressure device be discharged into normal pressure and ejection in the moment of valve opening; The said opening time is about 2-3 second, and the closing presure valve recovers the temperature and pressure in the heat preservation pressure device then; Subsequently; Every be preferably 7-10 second at a distance from 5-12 second, pressure valve can be opened so that the raw material of the part cellulose in the heat preservation pressure device is discharged into normal pressure automatically; Automatically close then, to realize the continuous steam explosion of fibrous raw material.
Said not special qualification of amount that at every turn is discharged into non-pressurized fibrous raw material; The specification of the heat preservation pressure device that the different production scale is adopted is different; The processing power of each steam explosion is also different; Therefore, can select the device of different size for use according to the production needs, thus the amount of the each cellulose containing raw material of handling of decision.According to the present invention, under the preferable case, the raw material that at every turn is discharged into non-pressurized cellulose is the 6-15 weight % of cellulose containing raw material in the heat preservation pressure device, more preferably 8-12 weight %.Generally, the raw material that at every turn is discharged into non-pressurized cellulose is the 120-180 gram, is preferably the 120-150 gram.Generally speaking, the raw material of said cellulose is at the uniform velocity to carry, and therefore, the transfer rate of the raw material of cellulose remains unchanged the amount of the raw material of the cellulose in this heat preservation pressure device basically.
Under the preferable case; For enhance productivity and can guarantee simultaneously steam explosion products and enzyme thorough mixing, obtain the enzyme speed of action of higher steam explosion products concentration and higher level; With the amount of first steam explosion products of enzyme blended be the 20-60% of whole steam explosion products weight, more preferably 20-40%; The amount of the steam explosion products that adds after first is the 40-80% of whole steam explosion products weight, more preferably 60-80%.
More preferably under the situation, said number of times in batches is 3 times, with the amount of first steam explosion products of enzyme blended be the 20-60% of whole steam explosion products weight, more preferably 20-40%; The amount of the steam explosion products of second batch of adding is the 20-50% of whole steam explosion products weight, more preferably 20-40%; The amount of the steam explosion products of the 3rd batch of adding is the 20-40% of whole steam explosion products weight, more preferably 20-30%.
In order to guarantee that enzyme has best reactive behavior; With first steam explosion products with before enzyme mixes, the pH value of steam regulation explosion product is greater than 3 less than 7, is preferably 4-5.5; So that steam explosion products is with after enzyme mixes, enzyme has best reactive behavior.The method of said conditioned reaction material pH value can adopt and well known to a person skilled in the art the whole bag of tricks, like the pH value according to the gained steam explosion products, in this steam explosion products, adds acidic substance or alkaline matter.For example, said acidic substance can be one or more in sulfuric acid, hydrochloric acid and the phosphoric acid; Said alkaline matter can be sodium hydroxide and/or Pottasium Hydroxide.
Under the preferable case, also comprise the timed interval between said two adjacent batch mixing steps.In order to make fully hydrolysis steam explosion products of enzyme, improve the transformation efficiency of sugar, the said timed interval is preferably 10-120 minute, more preferably 30-60 minute.
In order to make enzyme and reaction raw materials blended more even, the enzymolysis of said steam explosion products preferably carries out under the condition that water exists, and more preferably in the suspension-s that contains water and steam explosion products, carries out.That is to say; When the present invention preferably mixes first steam explosion products with enzyme, add entry, first steam explosion products and water are mixed with suspension-s after; Again the required whole enzymes of enzymolysis are mixed with suspension-s, and then continue to add steam explosion products in batches according to method of the present invention.Do not limit the amount of the water that added is special, under the preferable case, the add-on of said water is 1-4 with whole weight ratios of steam explosion products: 1.More preferably under the situation; Mixture to obtain after first steam explosion products, water and the enzyme mixing is a benchmark, and with the dry weight basis of steam explosion products, the content of first steam explosion products is at least 1 weight %; Be preferably 1-20 weight %, more preferably 1.2-10 weight %.In existing feedstock production alcoholic acid method by cellulose, when enzymolysis,, cause the production cycle long because the concentration of the raw material of cellulose is too rare, production efficiency is low.And contriver of the present invention finds; The concentration of steam explosion products in the mixture of when enzymolysis, controlling first steam explosion products, water and obtaining after enzyme mixes, with first steam explosion products, water and the mixture that obtains after enzyme mixes be benchmark, with the dry weight basis of steam explosion products; Make the content of steam explosion products be at least 1 weight %; Be preferably 1-20 weight %, more preferably 1.2-10 weight % can either guarantee that the cycle of enzymolysis is short; Guarantee to have high production efficiency simultaneously, promptly the productive rate of monose is high.
In addition, when adopting prior art to carry out the raw material of steam blasting cellulose-containing, the raw material that it has been generally acknowledged that said cellulose is corn straw for example; Fully water logging; Can obtain the effect of better steam explosion, because abundant water logging can make the raw material of said cellulose, for example a large amount of water is contained in corn straw inside; When steam explosion; The raw material of the said cellulose for example inner water of corn straw can be gasificated into water vapor, is for example gone out corn straw inside by the raw material of said cellulose during pressure release, with the raw material that fully the destroys said cellulose from inside to outside xylogen sheath of corn straw embedding Mierocrystalline cellulose and semicellulose for example; Contriver of the present invention is surprised to find that; The water cut of raw material that control is used for the cellulose of steam explosion is 30-60 weight %, can further guarantee under the steam explosion condition, to make the inner aqueous vapor of raw material of cellulose to change into water vapor, helps when pressure release water vapour and is gone out by the raw material inside of cellulose; With the xylogen sheath of embedding Mierocrystalline cellulose and semicellulose in the raw material of abundant destruction cellulose, thereby further improve ethanol yield.According to the present invention, the raw material of the cellulose of water cut all is an available arbitrarily, and under the preferable case, the water cut of the raw material of the used cellulose of the present invention is 30-60 weight %, more preferably 35-50 weight %.
According to the present invention, can obtain the raw material that water cut is the cellulose of 30-60 weight % through variety of way.The raw material of the different batches cellulose that there are differences for water cut can carry out different processing, such as, for the high stalk of water cut, can make its water cut be reduced to 30-60 weight % at 45-90 ℃ of oven dry part moisture; For the low stalk of water cut, can adopt control water logging time method, the water cut that increases stalk is to 30-60 weight %; Just in time at stalk, can directly carry out steam explosion for water cut to 30-60 weight %.The water cut of the raw material of the preferred said cellulose that is used for steam explosion is 35-50 weight %.Only if specified otherwise; The water cut of the raw material of cellulose according to the invention all is meant the initial weight W1 of cellulose containing raw material and under 70-100 ℃, dries poor to the dried meter weight W 2 of the cellulose containing raw material of constant weight; With the weight percent of the initial weight W1 of cellulose containing raw material, i.e. water cut (weight %)=(W1-W2)/W1 * 100%.
Under the preferable case; Continuous processing for the raw material of realizing cellulose; Can drench carrying out water in the continuous conveying process before the raw material of cellulose is in being delivered to the heat preservation pressure device continuously; The water consumption that water drenches makes the water cut of the raw material of cellulose preferably remain on 30-60 weight %, is preferably 35-50 weight %, and then is delivered to continuously and carries out steam explosion in the heat preservation pressure device.
Because the present invention only relates to the improvement of the feed way of the steam explosion products that obtains to the method for steam explosion and by the raw material of steam blasting cellulose-containing, therefore other step with the feedstock production alcoholic acid method of cellulose is had no particular limits.
The enzyme that said enzymolysis uses comprises cellulase.Said cellulase can obtain through variety of way, for example is commercially available, perhaps through using the microbes producing cellulase secretion to obtain.
Because the enzyme that uses the microbes producing cellulase secretion to obtain can contain various by products, the therefore preferred enzyme that directly adds.The consumption of said enzyme is The more the better, and from cost consideration, preferably with the dry weight basis of every gram steam explosion products, the consumption of said cellulase is the 8-20 enzyme activity unit, more preferably the 10-15 enzyme activity unit.The enzyme activity of cellulase according to the invention is according to American National renewable energy source laboratory (NationalRenewable Energy Laboratory; NREL) standard method that provides---cellulase activity is measured NREL LAP-006 and is measured; The enzyme activity unit of said cellulase is under the condition determination of this standard method regulation, restrains the micrograms that Whatman No.1 filter paper is converted into the required enzyme of glucose with 1 in 1 minute.
The temperature of said enzymolysis can be any optimum temperature of cellulase, is generally 45-55 ℃, more preferably 48-52 ℃.The longer the better on the time theory of said enzymolysis, considers plant factor, and the time of preferred said enzymolysis is 25-48 hour, more preferably 30-40 hour.Said enzymolysis time begins to calculate from the raw materials mix of enzyme and cellulose the time, that is, said enzymolysis time refer to the raw material with cellulose mixes with enzyme beginning to the feedstock conversion of cellulose for sugar after sugared content no longer increase the used time.Therefore, according to the present invention, the timed interval between said adjacent two batch mixing steps is also included within the said enzymolysis time.
Said cellulase is a prozyme, comprises C at least 1The plain enzyme of fiber type, the plain enzyme of Cx fiber type and three kinds of enzymes of cellose.
C 1Enzyme can make the crystalline Mierocrystalline cellulose change amorphous Mierocrystalline cellulose into.
C xThe plain enzyme of fiber type is divided into C again X1Plain enzyme of fiber type and C X2Two kinds of the plain enzymes of fiber type.C X1The plain enzyme of fiber type is the endo-type cellulase, can be from the plain intramolecular action of hydration amorphous cellulose in β-1, and the 4-glycosidic link generates cellodextrin and cellobiose.C X2The plain enzyme of fiber type is a kind of circumscribed-type cellulase, can act on β-1 from the non-reducing end of hydration non-cellulose molecule, and the 4-glycosidic link cuts off β-1 one by one, and the 4-glycosidic link generates glucose.
Cellose then acts on cellobiose, generates glucose.
Under the preferable case, the enzyme that said enzymolysis uses also comprises hemicellulase.Because hemicellulase can make semicellulose be degraded into water-soluble wood sugar; So the enzyme that enzymolysis uses comprises hemicellulase; Exposed fibers is plain more fully on the one hand; Increase the contact probability of Mierocrystalline cellulose and cellulase, semicellulose degraded product wood sugar can be generated ethanol by the pichia stipitis fermentation on the other hand, and two aspect effects can increase alcohol yied.With the dry weight basis of every gram steam explosion products, the consumption of said hemicellulase is the 4.4-8.8 enzyme activity unit.
The enzyme activity unit of hemicellulase according to the invention (U) is under 50 ℃, pH=4.8 condition, and it is that 1 weight % xylan solution produces the required enzyme amount of 1 microgram reducing sugar (in wood sugar) that PM decomposes concentration.
The vigor of hemicellulase according to the invention refers to the unit of activity that every gram hemicellulase is had.It is that hydrolysis 1 weight % xylan produces reducing sugar (in wood sugar) under 4.8 the condition at 50 ℃, pH that the vigor of said hemicellulase utilizes hemicellulase; Gained reducing sugar and excessive 3; Color reaction takes place in 5-dinitrosalicylic acid (DNS), records the proportional mensuration of growing amount of the absorbance value and the reducing sugar (in wood sugar) of reaction solution 550 nanometers with spectrophotometer.Concrete measuring method is following:
Accurately take by weighing 1.000 gram xylans, dissolve, use deionized water constant volume to 100 milliliter then, obtain 1 weight % xylan solution with the 0.1 mol acetate of 0.5 milliliter of pH=4.8-sodium acetate buffer solution;
Take by weighing 30 the gram Rochelle salts put into 500 milliliters of Erlenmeyer flasks, add 16 the gram NaOH after, add 50 ml deionized water; Speed heating in water bath to solid matter with 5 ℃/minute dissolves, and adds 1 gram 3, the 5-dinitrosalicylic acid; To dissolving, be cooled to room temperature, be settled to 100 milliliters with deionized water; Can get 3,5-dinitrosalicylic acid (DNS) solution;
Wood sugar is dried to constant weight for 80 ℃, accurately take by weighing 1.000 grams and be dissolved in 1000 ml waters, it is anticorrosion to add 10 milligrams of sodiumazide, obtains the normal wood sugar soln of 1 mg/ml;
Accurately take by weighing 1.000 gram solid hemicellulases or pipette 1 milliliters of liquid hemicellulase stoste; 0.1 mole of acetic acid-sodium acetate buffer solution dissolving with 0.5 milliliter of pH=4.8; Use deionized water constant volume to 100 milliliter then, obtain diluting 100 times enzyme liquid to be measured;
Respectively will be in 50 ℃ of heating in water bath 2 milliliters of wood sugar gradient standardized solution (0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml and 0..5 mg/ml of 60 minutes; Said wood sugar gradient standardized solution is mixed with the normal wood sugar soln of deionized water and 1 mg/ml) or deionized water (wood sugar blank); With 2 milliliters of DNS mixing boiling water baths 5 minutes; After the cooling, 15 milliliters of deionized water constant volumes, with the absorbance value of spectrophotometer wood sugar gradient standardized solution after distinguishing assaying reaction under 550 nanometers; With the absorbance value is X-coordinate, and xylose concentration is an ordinate zou drawing standard curve.Can get regression equation y=bx+a by this typical curve, wherein, x is an absorbance value, and y is an xylose concentration, and a is the intercept of gained straight-line equation, and b is the slope of gained straight-line equation;
Get 0.1 mol acetate-sodium acetate buffer solution (xylan blank) of 0.2 milliliter of enzyme liquid to be measured and 1.8 milliliters of said 1 weight % xylan solutions or pH=4.8, according to the step test light absorption value identical with above-mentioned wood sugar gradient standardized solution.And according to the vigor of computes hemicellulase:
Figure S2008101013145D00131
X is the absorbance value of enzyme liquid to be measured in the formula; B is consistent to b in the regression equation of absorbance value and a with xylose concentration with a; N is the extension rate of enzyme, 60 be expressed as enzymatic reaction time be 60 minutes, 5 for the sampling multiple (from 1 milliliter of enzyme liquid to be measured, having taken out 0.2 milliliter here tests).
Can determine the vigor of concrete hemicellulase according to aforesaid method, and then calculate the consumption of hemicellulase.
Can ferment pentoses and/or the mikrobe of hexose may be used to fermenting process of the present invention because yeast saccharomyces cerevisiae is the mikrobe of the zymohexose that ethanol-tolerant, by product are few, alcohol yied is high of widespread usage on the wine industry; Pichia stipitis for both can ferment pentoses also can zymohexose mikrobe (referring to " pichia stipitis continuously ferment pentose and hexose generate alcohol "; Ji Gengsheng etc.; Nanjing Forestry University's journal natural science edition, the 28th the 3rd phase of volume, 9-13 page or leaf; 2004), therefore the employed yeast of preferred said fermentation is pichia stipitis and/or yeast saccharomyces cerevisiae.In every gram enzymolysis product, the employed zymic inoculum size of said fermentation is 10 3-10 8Colony-forming unit, more preferably 10 4-10 6Colony-forming unit.Employed yeast is fermented in the present invention can be for being purchased yeast solids preparation (such as dried yeast powder) or barms (such as the cereuisiae fermentum of ATCC numbering 2601).Said zymic colony-forming unit can be measured by means commonly known in the art, such as the methylene blue staining viable bacteria counting method.The concrete grammar of methylene blue staining viable bacteria counting method is following:
1 gram dried yeast powder is dissolved in 10 ml sterile waters, or 1 milliliter of actication of culture liquid is diluted to 10 milliliters with sterilized water, add 0.5 milliliter of 0.1 weight % methylene blue, be incubated 30 minutes down at 35 ℃.Under 10 times of opticmicroscopes,, can get the number of viable bacteria in 1 gram dry yeast or the 1 milliliter of actication of culture liquid, i.e. colony forming single-digit with the number (dead bacterium dyeing, viable bacteria is not dyeed) of viable bacteria in the solution after the blood counting chamber counting insulation.
Said yeast can adopt conventional method inoculation, for example in enzymolysis product, adds the seed liquor of 5-15 volume %.Said seed liquor can be the aqueous solution or the culture medium solution of dry yeast, also can or be purchased the activated seed liquid of bacterial classification for dry yeast.The temperature of said fermentation can be any temperature that is suitable for yeast growth, is preferably 30-36 ℃, more preferably 32-35 ℃.The time of said fermentation can be for beginning from inoculation to occur to the decline phase of yeast growth the time of (being that fermentation time is that lag phase, logarithmic phase add stationary phase), and the time of preferred fermentation is 32-48 hour, more preferably 32-40 hour.Tunning ethanol can be used conventional method, according to requirement (requiring alcoholic acid purity to reach more than 99% such as the fuel alcohol) separation and refining of different Industrial products, such as distilling, concentrate, dewatering.
In addition; The pressure and temperature of steam explosion can be sterilized; For preventing that in enzymolysis process assorted bacterium (mainly being bacterium) pollutes the toxin of the enzyme activity that exerts an influence; And prevent that living contaminants influences the zymic growth during the fermentation, and preferably before enzymolysis, adding bacteria antibiotic such as industrial penicillium mould, said microbiotic does not have effect to yeast but the growth of the bacterium that can suppress to mix.With every milliliter of enzymolysis solution or fermented liquid is benchmark, and said antibiotic add-on is a 1-10 unit.Said enzymolysis solution comprises steam explosion products, enzyme and water, and said fermented liquid comprises the yeast of enzymolysis product and inoculation.
Because under the high-temperature and high-pressure conditions of steam explosion; Semicellulose in the raw material of cellulose can produce the inhibition such as furfural, furans; Said inhibition can reduce the vigor and the zymogenic activity of enzyme; Therefore said method also is included in after the raw material of steam blasting cellulose-containing, before the steam explosion products that enzymolysis obtains, washes said steam explosion products.Said washing is that steam explosion products is mixed with water, stirs, and above-mentioned inhibition is dissolved in the water, spinning steam explosion products then, thus from steam explosion products, remove inhibition.The temperature of said washing is high more in theory, and is can the dissolved inhibition many more, considers energy consumption, and the temperature of preferred said washing is at least 60 ℃, more preferably 60-80 ℃.Because the water that added of washing is many more, the solvent that then dissolves inhibition is many more, but considers the energy consumption of spinning, and preferably with the dry weight basis of the raw material of every gram cellulose, the amount of water of said washing is the 2-10 gram, more preferably the 2-3 gram.
Owing to may contain sandstone impurity and iron contamination in the raw material of cellulose; Can cause damage to steam explosion equipment; So preparation alcoholic acid method of the present invention; Can be included in steam explosion and before the raw material of cellulose removed stone deironing routine operation, such as removing the stone deironing with the raw material of " wind send " cellulose and the method for attraction.Sandstone can not be delivered in the steam explosion equipment by wind because quality is big, iron contamination since the attraction of magnet can not get in the steam explosion equipment with raw material yet, thereby can accomplish except that the stone deironing.In addition; Because the raw material of cellulose itself tangles and the occluding device pipeline easily; Therefore; Before getting into steam explosion equipment, the size that preferably makes the raw material of said cellulose is 0.2-1 centimetre of a 0.5-3 cm x 0.2-1 cm x, and the size that more preferably makes the raw material of said cellulose is 0.5-1 centimetre of a 1-2 cm x 0.4-0.6 cm x.
The present invention will be described in more detail below in conjunction with embodiment.
Embodiment 1
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
(1) continuous steam explosion
Test corn straw raw material as shown in Figure 1; The water cut that records corn straw according to above-mentioned method is 20 weight %; Raw material is cut into the segment of 0.5 centimetre of 1.5 cm x, 0.2 cm x; With this water cut is that the corn straw section of 20 weight % is sent into from feeder continuously, and carries out water before in being delivered to the heat preservation pressure device and drench, and water drenches that to make the water cut of this corn straw section be 40 weight %.Be that the corn straw of 40 weight % is delivered in the heat preservation pressure device with 2000 these water cut of gram continuously then, the pressure of said heat preservation pressure device is 1.6 MPas, and temperature is 195 ℃; Said corn straw transmits in this heat preservation pressure device continuously, and the LV of transmission makes this corn straw under 195 ℃, 1.6 MPas, to keep 5 minutes, afterwards; Every at a distance from 10 seconds, the valve opening that makes this heat preservation pressure device once, after 2 seconds; Valve closes; Make the ejection of part corn straw, be discharged into normal pressure, accomplish steam explosion, and repeat said process and carry out continuous steam explosion.The raw material that at every turn is discharged into non-pressurized cellulose is 11 weight % of cellulose containing raw material in the heat preservation pressure device, about 165 grams.Collect steam explosion products at last; And the water of gained steam explosion products and 70 ℃ mixed 30 minutes according to weight ratio at 1: 3; Use LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out solid-liquid separation then, obtain 2700 gram solid steam explosion products (water cut is 55.6 weight %) altogether.
The mensuration of Mierocrystalline cellulose gross weight and semicellulose gross weight in the gained solid steam explosion products:
Get the steam explosion products of 5 grams after the said washing and under 45 ℃, dry to constant weight 2.22 grams, 300.0 milligrams of this dried steam explosion products of weighing are positioned in heavy 80 100 milliliters of dry Erlenmeyer flasks restraining.Add the sulphuric acid soln that 3.00 ml concns are 72 weight % to said Erlenmeyer flask, stirred 1 minute.Then Erlenmeyer flask was heated 60 minutes in 30 ℃ of water-baths, whenever stirred once to guarantee even hydrolysis at a distance from 5 minutes.Water-bath makes the vitriolic concentration dilution to 4 weight % with deionized water after finishing.Filter the steam explosion products acid hydrolysis liquid after the said dilution with B, obtain altogether filtrating 84 milliliters.20 milliliters of filtratings are transferred in the triangular flask of 50 milliliters of exsiccant.Use 2.5 gram lime carbonate to regulate this pH value of filtrate to 5.5, left standstill 5 hours, collect supernatant liquid.0.2 the supernatant liquid that the micron membrane filtration is collected, the filtrating of gained supernatant liquid is analyzed with Biorad Aminex HPX-87P performance liquid chromatography (HPLC).HPLC condition: sample size 20 microlitres; Moving phase is 0.2 micron membrane filtration, and the HPLC ultrapure water of the sonic oscillation degassing; Flow velocity is 0.6 ml/min; Column temperature 80-85 ℃; Detector temperature 80-85 ℃; Detector is a refractive index detector; Be 35 minutes working time.With D-(+) glucose of 0.1-4.0 mg/ml concentration range and 0.1-4.0 mg/ml concentration range D-(+) wood sugar as standard model.HPLC analyzes and to obtain that glucose concn is 2.59 mg/ml in the steam explosion products acid hydrolysis liquid; The steam explosion products acid hydrolysis that calculating can get after the said washing of 1 gram can obtain the glucose that weight is 0.218 gram; Because being the sulphuric acid soln of 72 weight %, concentration can the Mierocrystalline cellulose of the product of steam explosion all be hydrolyzed into glucose; Therefore the weight of gained glucose is 1.11 times of cellulosic weight in the steam explosion products; Cellulose 0.196 gram in the steam explosion products after the i.e. said washing of 1 gram then is total to cellulose 528 grams in the 2700 gram steam explosion products.HPLC analyzes and to obtain that xylose concentration is 0.73 mg/ml in the steam explosion products acid hydrolysis liquid; The steam explosion products acid hydrolysis that calculating can get after the said washing of 1 gram can obtain the wood sugar that weight is 0.061 gram; Because being the sulphuric acid soln of 72 weight %, concentration can semicellulose in the product of steam explosion all be hydrolyzed into wood sugar; Therefore the weight of gained wood sugar is 1.14 times of weight of semicellulose in the steam explosion products; Promptly contain semicellulose 0.054 gram in the 1 gram steam explosion products, then contain semicellulose 145 grams altogether in the 2700 gram steam explosion products.
(2) enzymolysis
In enzymatic vessel, add entry, and under agitation, add remaining washing steam explosion products behind first step (1) sampling and testing; The amount of the washing steam explosion products of first adding be behind step (1) sampling and testing remaining whole washing steam explosion products weight 35%; Regulating the pH value then is 4, be heated to 52 ℃ after, with the dry weight basis of every gram steam explosion products; The cellulase that adds 20 enzyme activity units calculates; All the dry weight of steam explosion products is 1200 grams, add the 24000 enzyme activity units cellulase (jade of the He family Bioisystech Co., Ltd) of (weight of enzyme is about 160 grams) altogether, and insulation mixes 30 minutes under 52 ℃; Add second batch of washing steam explosion products then, the amount of the washing steam explosion products of second batch of adding be behind step (1) sampling and testing remaining whole washing steam explosion products weight 35%, and 52 ℃ down insulation mixed 50 minutes; And then add the 3rd batch the washing steam explosion products; The amount of the washing steam explosion products of the 3rd batch of adding be behind step (1) sampling and testing remaining whole washing steam explosion products weight 30%, and 52 ℃ down the insulation enzymolysis obtain enzymolysis product after 38 hours.The add-on of said water is 2: 1 with the weight ratio of all washing steam explosion products.Enzymolysis product is filtered with B, 20 milliliters of filtratings are transferred in the triangular flask of 50 milliliters of exsiccant, left standstill 5 hours, collect supernatant liquid.0.2 the supernatant liquid that the micron membrane filtration is collected, according to the described performance liquid condition of above-mentioned steps (1), the glucose of measuring and calculating in the enzymolysis product 549.7 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 495.3 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11, and according to computes cellulose conversion rate and monose productive rate, calculation result is seen table 1.Per hour the cellulosic amount of enzymolysis is 12.6 grams (enzymolysis time is 39.3 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 6.5 weight %.)
Cellulose conversion rate=100% * by the cellulosic weight of enzymolysis/cellulosic gross weight
Glucose weight/stalk dry weight that monose productive rate=100% * enzymolysis obtains
(3) fermentation
Make the temperature of enzymolysis product reduce to 35 ℃, in the weight of every gram enzymolysis product, inoculation 10 5The yeast saccharomyces cerevisiae of colony-forming unit (the super highly active dry yeast in Angel, Hubei Angel Yeast stock company), gained mixture under 32 ℃ in fermentor tank stir culture 40 hours.At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 284.6 grams at 78.3 ℃ of following second distillations, and according to the computes alcohol yied, calculation result is seen table 1.
Alcohol yied=100% * ethanol weight/stalk dry weight
Embodiment 2
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Method according to embodiment 1 prepares ethanol; Different is; Corn straw section with water cut being 20 weight % is sent into from feeder continuously, and carries out the water pouring before in being delivered to the heat preservation pressure device, and it is 75 weight % that the water pouring makes the water cut of this corn straw section.Be that the corn straw of 75 weight % is delivered to and carries out continuous steam explosion in the heat preservation pressure device with 2000 these water cut of gram continuously according to the condition of embodiment 1 and method then, obtain 3000 gram solid steam explosion products (water cut is 83.3 weight %) altogether after collecting steam explosion products and washing.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 3000 gram steam explosion products cellulose 210 grams altogether, semicellulose 60 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and the consumption of enzyme is about 65 grams, after adding the 3rd batch of steam explosion products, is incubated enzymolysis down at 52 ℃ and obtains enzymolysis product after 25 hours.And according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 218.2 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 196.6 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Per hour the cellulosic amount of enzymolysis is 7.5 grams (enzymolysis time is 26.3 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 2.5 weight %.) second distillation can get ethanol 116.5 gram.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 3
Present embodiment explanation the present invention is with the feedstock production alcoholic acid method of cellulose.
Method according to embodiment 1 prepares ethanol; Different is; Be not carry out water after the corn straw section of 20 weight % is sent into continuously to drench directly to be delivered to and carry out continuous steam explosion in the heat preservation pressure device from feeder with 2000 gram water cut; The condition of steam explosion is collected steam explosion products and is also obtained 3556 gram solid steam explosion products (water cut is 55 weight %) altogether after the washing with embodiment 1.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 3556 gram steam explosion products cellulose 680 grams altogether, semicellulose 193.8 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and the consumption of enzyme is about 210 grams, after adding the 3rd batch of steam explosion products, is incubated enzymolysis down at 52 ℃ and obtains enzymolysis product after 40 hours.And according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 703.5 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 633.8 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Per hour the cellulosic amount of enzymolysis is 15.3 grams (enzymolysis time is 41.3 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 6.5 weight %.) second distillation can get ethanol 368.1 gram.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Comparative Examples 1
The feedstock production alcoholic acid reference method of cellulose is adopted in the explanation of this Comparative Examples.
Method according to embodiment 1 prepares ethanol, and different is that (Zhang Xuefeng, orchid are kindly helped secure the success of according to document " to the introduction of straw biomass to alcohol conversion process "; Wine brewing, the 29th the 4th phase of volume, 37-38 page or leaf; In July, 2002) disclosed method; Stalk was soaked in water earlier before steam explosion 40 minutes, and the water cut that records the corn straw raw material after the immersion reaches 83 weight %, and the corn straw of getting 2000 these water cut of gram then and be 83 weight % (every batch 1000 gram) in two batches carries out the intermittent type steam explosion; The method of said intermittent type steam explosion is for keeping 5 minutes pressure release then with the said corn straw of 1000 grams earlier with the pressure of 1.6 MPas under 195 ℃; And then the said corn straw of remaining 1000 grams carried out steam explosion under same condition, obtain 2300 gram steam explosion products (water cut is 85.2 weight %) altogether after collecting steam explosion products and washing.Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 2300 gram steam explosion products cellulose 142.8 grams altogether, semicellulose 45 grams.
Carry out enzymolysis and fermentation according to the method for embodiment 1 then; Different is; In enzymolysis step (2), once all join remaining washing steam explosion products behind step (1) sampling and testing in the aqueous enzymatic vessel; Mix (weight ratio of all washing steam explosion products and water is 2: 1) then with enzyme, enzymolysis time needs 40 hours.Enzymolysis product is filtered with B, 20 milliliters of filtratings are transferred in 50 milliliters of triangular flasks of exsiccant, left standstill 5 hours, collect supernatant liquid.0.2 the supernatant liquid that the micron membrane filtration is collected, according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 117.9 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 106.2 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11; Second distillation obtains ethanol 62.2 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Comparative Examples 2
The feedstock production alcoholic acid reference method of cellulose is adopted in the explanation of this Comparative Examples.
Method according to embodiment 1 prepares ethanol; Different is; With 1500 gram water cut is that 20 weight % corn straws were directly kept under the pressure with 1.8 MPas under 190 ℃ 4 minutes; Steam explosion is accomplished in pressure release then, collects steam explosion products and also obtains 3000 gram steam explosion products (water cut is 60 weight %) altogether after the washing.Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 3000 gram steam explosion products cellulose 494 grams altogether, semicellulose 146.6 grams.And steam explosion products is carried out enzymolysis according to the method for above-mentioned Comparative Examples 1; Enzymolysis time needs 46 hours; Enzymolysis product is fermented, and then according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 420.0 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 378.4 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11; Second distillation obtains ethanol 223.2 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
The electron microscopic observation result:
Amplify 1000 times of steam explosion products of observing Comparative Examples 1, Comparative Examples 2 and embodiment 1 with the JSM-35CF ESEM, the result is respectively like Fig. 2, Fig. 3 and shown in Figure 4.As can beappreciated from fig. 2, because stalk inside does not have moisture can be gasificated into water vapor, so steam explosion can not fully destroy stalk from inside to outside; As can beappreciated from fig. 3, the effect of Comparative Examples 2 corn straw steam explosions and bad, Mierocrystalline cellulose still is pencil, and cellulose surface is residual that the xylogen sheath arranged; And the steam explosion effect of embodiment 1 as shown in Figure 4 is fine, can see the free Mierocrystalline cellulose.
Embodiment 4
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Method according to embodiment 1 is carried out continuous steam explosion to corn straw; Different is; Water drenches that to make the water cut that gets into the corn straw before the heat preservation pressure device be 65 weight %, and the corn straw that is 65 weight % with this 2000 gram water cut continuously is delivered to and carries out continuous steam explosion in the heat preservation pressure device.The raw material that at every turn is discharged into non-pressurized cellulose is 8.5 weight % of cellulose containing raw material in the heat preservation pressure device, about 127.5 grams.Condition is identical with embodiment 1; Collect steam explosion products; And mixed at 1: 3 30 minutes according to weight ratio with 60 ℃ water; Use LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out solid-liquid separation then, obtain 2500 gram solid steam explosion products (water cut is 72 weight %) altogether.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 2500 gram steam explosion products cellulose 297.7 grams altogether, semicellulose 80.6 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and the consumption of enzyme is about 95 grams, after adding the 3rd batch of steam explosion products, is incubated enzymolysis down at 52 ℃ and obtains enzymolysis product after 27 hours.And according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 308.7 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 278.1 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Per hour the cellulosic amount of enzymolysis is 9.8 grams (enzymolysis time is 28.3 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 4.1 weight %.) second distillation can get ethanol 162.4 gram.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 5
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Method according to embodiment 1 is carried out continuous steam explosion to corn straw; Different is; Be after the corn straw of 20 weight % is cut into the segment of 0.5 centimetre of 2.5 cm x, 0.5 cm x earlier with water cut; With vertical steeping cell (the auspicious mechanical ltd in river in Shangdong Province, Shandong) and weight is after 1/3rd water of said corn straw raw material weight mixes, and can get the corn straw of water cut 50 weight %.The corn straw that is 50 weight % with this 2000 gram water cut continuously then is delivered in the heat preservation pressure device, and the pressure of said heat preservation pressure device is 1.5 MPas, and temperature is 185 ℃; Said corn straw transmits in this heat preservation pressure device continuously, and the LV of transmission makes this corn straw under 185 ℃, 1.5 MPas, to keep 7 minutes, afterwards; Every at a distance from 8 seconds, the valve opening that makes this heat preservation pressure device once, after 2 seconds; Valve closes; Make the part corn straw be discharged into the normal pressure ejection, accomplish steam explosion, and repeat said process and carry out continuous steam explosion.The raw material that at every turn is discharged into non-pressurized cellulose is 9.2 weight % of cellulose containing raw material in the heat preservation pressure device, about 138 grams.Collect steam explosion products at last, after with said continuous steam explosion, collect not totally 2250 gram steam explosion products (water cut is 55.5 weight %) through washing.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 2250 gram steam explosion products cellulose 426 grams altogether, semicellulose 118.5 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and the consumption of enzyme is about 130 grams, after adding the 3rd batch of steam explosion products, is incubated enzymolysis down at 52 ℃ and obtains enzymolysis product after 33 hours.And according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 434 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 391 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Per hour the cellulosic amount of enzymolysis is 11.4 grams (enzymolysis time is 34.3 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 6.5 weight %.) second distillation can get ethanol 226 gram.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 6
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
(1) continuous steam explosion
Getting 2000 gram water cut is the corn straw of 35 weight %, is cut into the segment of 0.7 centimetre of 1.2 cm x, 0.4 cm x, and the corn straw that is 35 weight % with this 2000 gram water cut continuously then is delivered in the heat preservation pressure device; The pressure of said heat preservation pressure device is 1.7 MPas, and temperature is 200 ℃, and said corn straw transmits in this heat preservation pressure device continuously; The LV that transmits makes this corn straw under 200 ℃, 1.7 MPas, to keep 4 minutes, and is afterwards, every at a distance from 8 seconds; The valve opening that makes this heat preservation pressure device once; After 2 seconds, valve closes makes the part corn straw be discharged into the normal pressure ejection; Accomplish steam explosion, and repeat said process and carry out continuous steam explosion.The raw material that at every turn is discharged into non-pressurized cellulose is 10 weight % of cellulose containing raw material in the heat preservation pressure device, about 150 grams.Collect steam explosion products at last; The water of gained steam explosion products and 75 ℃ was mixed 50 minutes according to weight ratio at 1: 2.5; Use impeller pump LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out solid-liquid separation then, obtain 5200 gram steam explosion products (water cut is 75 weight %) altogether.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 5200 gram steam explosion products cellulose 559 grams altogether, semicellulose 150 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and the consumption of enzyme is about 170 grams, after adding the 3rd batch of steam explosion products, is incubated enzymolysis down at 52 ℃ and obtains enzymolysis product after 40 hours.And according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 572 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 515.3 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Per hour the cellulosic amount of enzymolysis is 12.5 grams (enzymolysis time is 41.3 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 3.7 weight %.) second distillation can get ethanol 296.4 gram.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 7
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
(1) continuous steam explosion
Getting 2000 gram water cut is the corn straw of 55 weight %, is cut into the segment of 0.8 centimetre of 2.0 cm x, 0.6 cm x, and the corn straw that is 55 weight % with this 2000 gram water cut continuously then is delivered in the heat preservation pressure device; The pressure of said heat preservation pressure device is 1.55 MPas, and temperature is 190 ℃, and said corn straw transmits in this heat preservation pressure device continuously; The LV that transmits makes this corn straw under 190 ℃, 1.55 MPas, to keep 6 minutes, and is afterwards, every at a distance from 7 seconds; The valve opening that makes this heat preservation pressure device once; After 2 seconds, valve closes makes the part corn straw be discharged into the normal pressure ejection; Accomplish steam explosion, and repeat said process and carry out continuous steam explosion.The raw material that at every turn is discharged into non-pressurized cellulose is 8 weight % of cellulose containing raw material in the heat preservation pressure device, about 120 grams.Collect steam explosion products at last; The water of gained steam explosion products and 65 ℃ was mixed 40 minutes according to weight ratio at 1: 2; Use impeller pump LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out solid-liquid separation then, obtain 2500 gram steam explosion products (water cut is 64 weight %) altogether.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 2500 gram steam explosion products cellulose 388 grams altogether, semicellulose 99.5 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and the consumption of enzyme is about 120 grams, after adding the 3rd batch of steam explosion products, is incubated enzymolysis down at 52 ℃ and obtains enzymolysis product after 39 hours.And according to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 403.2 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 363.2 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Per hour the cellulosic amount of enzymolysis is 9.0 grams (enzymolysis time is 40.3 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 5.3 weight %.) second distillation can get ethanol 212.4 gram.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 8
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Is that the 1950 gram corn straws of 40 weight % carry out continuous steam explosion according to the method for embodiment 1 to said water cut, collects steam explosion products 3050 grams (water cut is 61.6 weight %) that obtain through washing at last.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 3050 gram steam explosion products cellulose 503 grams altogether, semicellulose 135 grams.
According to above-mentioned not the steam explosion products of the method enzymolysis of embodiment 1 through washing; Different is; After in enzymatic vessel, adding entry, and under agitation, steam explosion products is mixed with enzyme in three batches; The amount of the steam explosion products of first adding be behind the sampling and testing remaining whole steam explosion products weight 60%, 50 ℃ down insulation mixed 100 minutes; Add second batch of steam explosion products then, the amount of the steam explosion products of second batch of adding be behind the sampling and testing remaining whole steam explosion products weight 20%, 50 ℃ down insulation mixed 30 minutes; And then add the 3rd batch of steam explosion products, the amount of the steam explosion products of the 3rd batch of adding be behind the sampling and testing remaining whole steam explosion products weight 20%, 50 ℃ down the insulation enzymolysis obtain enzymolysis product after 37 hours, the consumption of enzyme is about 156 grams.According to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 517.1 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 465.9 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11, and per hour the cellulosic amount of enzymolysis is 11.9 grams (enzymolysis time is 39.2 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 8.7 weight %.)
Fermentation step is with embodiment 1, and second distillation obtains ethanol 269.1 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 9
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Is that the 1950 gram corn straws of 40 weight % carry out continuous steam explosion according to the method for embodiment 1 to said water cut; The raw material of cellulose carries out continuous steam explosion; The water of gained steam explosion products and 75 ℃ was mixed 50 minutes according to weight ratio at 1: 2.5; Use impeller pump LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out solid-liquid separation then, obtain 2950 gram washing solid steam explosion products (water cut is 60.3 weight %) altogether.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 2950 gram washing steam explosion products cellulose 503 grams altogether, semicellulose 136 grams.
The above-mentioned steam explosion products of method enzymolysis according to embodiment 1; Different is; After in enzymatic vessel, adding entry, and under agitation, will wash steam explosion products and mix with enzyme in three batches; The amount of the washing steam explosion products of first adding be behind the sampling and testing remaining whole washing steam explosion products weight 20%, 52 ℃ down insulation mixed 20 minutes; Add second batch of washing steam explosion products then, the amount of the washing steam explosion products of second batch of adding be behind the sampling and testing remaining whole washing steam explosion products weight 40%, 52 ℃ down insulation mixed 40 minutes; And then add the 3rd batch the washing steam explosion products; The amount of the washing steam explosion products of the 3rd batch of adding be behind the sampling and testing remaining whole washing steam explosion products weight 40%; Be incubated enzymolysis down at 52 ℃ and obtain enzymolysis product after 40 hours, the consumption of enzyme is about 156 grams.According to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 522.0 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 470.3 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11, and per hour the cellulosic amount of enzymolysis is 11.5 grams (enzymolysis time is 41 hours).(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is 3.5 weight %.)
(3) fermentation
According to the method fermenting enzyme hydrolysis products of embodiment 1, different is, makes the temperature of enzymolysis product reduce to 32 ℃, in the weight of every gram enzymolysis product, and inoculation 10 6The yeast saccharomyces cerevisiae of colony-forming unit (the super highly active dry yeast in Angel, Hubei Angel Yeast stock company), gained mixture under 32 ℃ in fermentor tank stir culture 36 hours; At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 272.6 grams at 78.3 ℃ of following second distillations.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 10
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Is that the 1950 gram corn straws of 40 weight % carry out continuous steam explosion according to the method for embodiment 1 to said water cut; The raw material of cellulose carries out continuous steam explosion; The raw material of cellulose carries out continuous steam explosion; The water of gained steam explosion products and 65 ℃ was mixed 40 minutes according to mass ratio at 1: 2; Use impeller pump LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out solid-liquid separation then, obtain 2950 gram solid steam explosion products (water cut is 60.3 weight %) altogether.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 2950 gram steam explosion products cellulose 503 grams altogether, semicellulose 136 grams.
The above-mentioned steam explosion products of method enzymolysis according to embodiment 1; Different is; After in enzymatic vessel, adding entry, and under agitation, will wash steam explosion products and mix with enzyme in three batches; The amount of the washing steam explosion products of first adding be behind the sampling and testing remaining whole washing steam explosion products weight 10%, 52 ℃ down insulation mixed 10 minutes; Add second batch of washing steam explosion products then, the amount of the washing steam explosion products of second batch of adding be behind the sampling and testing remaining whole washing steam explosion products weight 40%, 52 ℃ down insulation mixed 40 minutes; And then add the 3rd batch the washing steam explosion products; The amount of the washing steam explosion products of the 3rd batch of adding be behind the sampling and testing remaining whole washing steam explosion products weight 50%; Be incubated enzymolysis down at 52 ℃ and obtain enzymolysis product after 40 hours, the consumption of enzyme is about 156 grams.The add-on of said water is 4: 1 with the weight ratio of all washing steam explosion products.According to the said performance liquid condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 518.0 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 466.7 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11, and per hour the cellulosic amount of enzymolysis is 11.4 grams (enzymolysis time is 40.8 hours).(with the dry weight basis of cellulosic material, the content of the raw material of first cellulose is 0.95 weight %.)
Fermentation step is with embodiment 1, and second distillation obtains ethanol 270.3 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, calculation result is seen table 1.
Embodiment 11
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Is that the corn straw of 40 weight % carries out continuous steam explosion according to the method for embodiment 1 to 1950 gram water cut; The water of gained steam explosion products and 65 ℃ was mixed 40 minutes according to mass ratio at 1: 2; Use impeller pump LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out solid-liquid separation then, obtain 2950 gram solid steam explosion products (water cut is 60.3 weight %) altogether.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 2950 gram steam explosion products cellulose 503 grams altogether, semicellulose 136 grams.
According to the above-mentioned steam explosion products through washing of the method enzymolysis of embodiment 1, different is, after in enzymatic vessel, adding entry; And under agitation; Add remaining washing steam explosion products behind first step (1) sampling and testing, the amount of the washing steam explosion products of first adding be behind step (1) sampling and testing remaining whole washing steam explosion products weight 60%, the pH value of conditioned reaction material is 5 then; And after being heated to 50 ℃; With the dry weight basis of every gram steam explosion products, add the cellulase of 15 enzyme activity units, the hemicellulase of 5 enzyme activity units calculates; All the dry weight of steam explosion products is 1170 grams; Add the cellulase (jade of the He family Bioisystech Co., Ltd) of 17550 enzyme activity units (about 110 grams) and the hemicellulase (Beijing chemical reagents corporation) of 5850 enzyme activity units (about 79 grams) altogether, the add-on of said water is 2.5: 1 with the weight ratio of all washing steam explosion products, and insulation mixed 80 minutes; Add then second batch the washing steam explosion products, the amount of the washing steam explosion products of second batch of adding be behind step (1) sampling and testing remaining whole washing steam explosion products weight 40%, the insulation enzymolysis obtain enzymolysis product after 38 hours.And according to the said performance liquid condition of embodiment 1 step (1); The glucose of measuring and calculating in the enzymolysis product 521.5 restrains totally; The weight of the glucose that said enzymolysis obtains is divided by 1.11; Be totally 469.8 to be restrained by the cellulosic weight of enzymolysis in the steam explosion products, the wood sugar of measuring and calculating in the enzymolysis product 111.9 restrains totally, and the weight of the wood sugar that said enzymolysis obtains is divided by 1.14; Be totally 98.2 to be restrained by the weight of the semicellulose of enzymolysis in the steam explosion products, per hour the gross weight of the Mierocrystalline cellulose of enzymolysis and semicellulose is 14.5 grams (enzymolysis time is 39.3 hours); And calculate Mierocrystalline cellulose, semicellulose transformation efficiency and monose productive rate according to the following equation, calculation result is seen table 1.(with the dry weight basis of steam explosion products, be benchmark with the blended weight that obtains, the content of said first steam explosion products is 7.5 weight %.)
Cellulose conversion rate=100% * by the cellulosic weight of enzymolysis/cellulosic gross weight
Semicellulose transformation efficiency=100% * by the gross weight of the weight/semicellulose of the semicellulose of enzymolysis
Monose productive rate=100% * (the wood sugar weight that glucose weight+enzymolysis that enzymolysis obtains obtains)/stalk dry weight
(3) fermentation
Make the temperature of enzymolysis product reduce to 34 ℃, in the weight of every gram enzymolysis product, inoculation is 10 4The yeast saccharomyces cerevisiae of colony-forming unit (the super highly active dry yeast in Angel, Hubei Angel Yeast stock company) and 10 4The pichia stipitis of colony-forming unit, gained mixture under 33 ℃ in fermentor tank stir culture 37 hours.At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 330.5 grams at 78.3 ℃ of following second distillations, and calculates alcohol yied according to the formula of embodiment 1, and calculation result is seen table 1.
Embodiment 12
Present embodiment is used to explain the feedstock production alcoholic acid method of employing cellulose of the present invention.
Is that the corn straw of 40 weight % is delivered in the heat preservation pressure device according to the method for embodiment 1 with 2000 these water cut of gram, and the raw material of said cellulose is carried out continuous steam explosion, obtains 3000 gram solid steam explosion products (water cut is 60 weight %) altogether.
Analyze the method for the steam explosion products of sulphuric acid hydrolysis according to the said HPLC of embodiment 1, record in the 3000 gram steam explosion products cellulose 528 grams altogether, semicellulose 145 grams.
Method according to embodiment 11 is carried out enzymolysis to steam explosion products; Different is; After in enzymatic vessel, adding entry, and under agitation, steam explosion products is mixed with enzyme in three batches; The amount of the steam explosion products of first adding be behind the sampling and testing remaining whole steam explosion products weight 40%, 50 ℃ down insulation mixed 50 minutes; Add second batch of steam explosion products then, the amount of the steam explosion products of second batch of adding be behind the sampling and testing remaining whole steam explosion products weight 30%, 50 ℃ down insulation mixed 30 minutes; And then add the 3rd batch of steam explosion products, the amount of the steam explosion products of the 3rd batch of adding be behind the sampling and testing remaining whole steam explosion products weight 30%, 50 ℃ down the insulation enzymolysis obtain enzymolysis product after 40 hours.And; Dry weight basis with every gram steam explosion products; Add the cellulase of 20 enzyme activity units, the hemicellulase calculating of 8 enzyme activity units; All the dry weight of steam explosion products is about 1200 grams, adds the cellulase (jade of the He family Bioisystech Co., Ltd) of 24000 enzyme activity units (about 160 grams) and the hemicellulase (Beijing chemical reagents corporation) of 9600 enzyme activity units (about 110 grams) altogether.And according to the said performance liquid condition of embodiment 1 step (1); The glucose of measuring and calculating in the enzymolysis product 550.9 restrains totally; The weight of the glucose that said enzymolysis obtains is divided by 1.11; Be totally 496.3 to be restrained by the cellulosic weight of enzymolysis in the steam explosion products, the wood sugar of measuring and calculating in the enzymolysis product 119.8 restrains totally, and the weight of the wood sugar that said enzymolysis obtains is divided by 1.14; Be totally 105.1 to be restrained by the weight of the semicellulose of enzymolysis in the steam explosion products, per hour the gross weight of the Mierocrystalline cellulose of enzymolysis and semicellulose is 14.6 grams (enzymolysis time is 41.3 hours); And calculate Mierocrystalline cellulose, semicellulose transformation efficiency and monose productive rate according to the method for embodiment 11, calculation result is seen table 1.(with the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of said first steam explosion products is 5.4 weight %.)
Method according to embodiment 5 is fermented to enzymolysis product, and the gained distillation fraction can get ethanol 342.7 grams at 78.3 ℃ of following second distillations, and calculates alcohol yied according to the formula of embodiment 1, and calculation result is seen table 1.
Table 1
Embodiment or Comparative Examples Cellulose conversion rate (%) Semicellulose transformation efficiency (%) Monose productive rate (%) Alcohol yied (%)
Embodiment 1 93.8 - 45.8 23.7
Comparative Examples 1 74.4 - 34.7 18.3
Comparative Examples 2 76.6 - 35.0 18.6
Embodiment 2 93.6 - 43.6 23.3
Embodiment 3 93.2 - 43.9 23.0
Embodiment 4 93.4 - 44.1 23.2
Embodiment 5 91.8 - 43.4 22.6
Embodiment 6 92.2 - 44.0 22.8
Embodiment 7 93.6 - 44.8 23.6
Embodiment 8 92.6 - 44.2 23.0
Embodiment 9 93.5 - 44.6 23.3
Embodiment 10 92.8 - 44.3 23.1
Embodiment 11 93.4 72.2 54.1 28.2
Embodiment 12 94.0 72.5 56.0 28.6
Data from last table 1 can find out, adopt the ethanol yield that the feedstock production alcoholic acid method of cellulose of the present invention obtains and the productive rate of monose and the method for Comparative Examples 1 and Comparative Examples 2 more all to be significantly improved.

Claims (17)

1. feedstock production alcoholic acid method that adopts cellulose, this method comprises the raw material of steam blasting cellulose-containing; With the steam explosion products that obtains mix with enzyme, enzymolysis; The enzymolysis product that fermentation obtains; It is characterized in that; The method of the raw material of said steam blasting cellulose-containing comprises that the raw material with cellulose is delivered in the heat preservation pressure device continuously, and the raw material of cellulose was kept in this heat preservation pressure device after 3-10 minute, and every separated 5-12 makes the raw material of the part cellulose in the said heat preservation pressure device be discharged into normal pressure second; Wherein, the raw material that at every turn is discharged into non-pressurized cellulose is the 6-15 weight % of the raw material of cellulose in the heat preservation pressure device; Mix steam explosion products said steam explosion products is comprised with the enzyme method of mixing with enzyme in batches, said number of times in batches is at least 2 times, and the amount of the steam explosion products of every batch of adding is at most 60% of whole steam explosion products weight.
2. method according to claim 1, wherein, the raw material of cellulose kept 4-7 minute in this heat preservation pressure device, and every separated 7-10 makes the raw material of the part cellulose in the said heat preservation pressure device be discharged into normal pressure second.
3. method according to claim 1 and 2, wherein, the raw material that at every turn is discharged into non-pressurized cellulose is the 120-180 gram.
4. method according to claim 1, wherein, the temperature of said heat preservation pressure device is 180-200 ℃, pressure is the 1.4-2.0 MPa.
5. method according to claim 1; Wherein, Said and amount first steam explosion products of enzyme blended is the 20-60% of whole steam explosion products weight, and the amount of the steam explosion products that adds after first is the 40-80% of whole steam explosion products weight.
6. method according to claim 1, wherein, said number of times in batches is 3 times, with the amount of first steam explosion products of enzyme blended be the 20-60% of whole steam explosion products weight; The amount of the steam explosion products of second batch of adding is the 20-50% of whole steam explosion products weight; The amount of the steam explosion products of the 3rd batch of adding is the 20-40% of whole steam explosion products weight.
7. according to claim 1,5 or 6 described methods, wherein, when first steam explosion products is mixed with enzyme; This method also comprises and adds entry; With the dry weight basis of steam explosion products, be benchmark with the weight of the mixture that obtains, the content of first steam explosion products is at least 1 weight %.
8. method according to claim 7 wherein, with the dry weight basis of steam explosion products, is a benchmark with the weight of the mixture that obtains, and the content of first steam explosion products is 1.2-10 weight %.
9. method according to claim 1, wherein, the water cut of the raw material of said cellulose is 30-60 weight %.
10. method according to claim 9, wherein, the water cut of the raw material of said cellulose is 35-50 weight %.
11. method according to claim 1 wherein, also comprises the timed interval between two adjacent batch mixing steps.
12. method according to claim 11, wherein, the said timed interval is 10-120 minute.
13. method according to claim 1, wherein, the enzyme that said enzymolysis uses comprises cellulase, and with the dry weight basis of every gram steam explosion products, the consumption of said cellulase is the 8-20 enzyme activity unit; The temperature of said enzymolysis is 45-55 ℃, and the time of said enzymolysis is 25-48 hour, and the pH value of said enzymolysis is 3.0-7.0.
14. method according to claim 13, wherein, the enzyme that said enzymolysis uses also comprises hemicellulase, and with the dry weight basis of every gram steam explosion products, the consumption of said hemicellulase is the 4.4-8.8 enzyme activity unit.
15. method according to claim 1, wherein, the employed yeast of said fermentation is pichia stipitis and/or yeast saccharomyces cerevisiae; In every gram enzymolysis product, the employed zymic inoculum size of said fermentation is 10 3-10 8Colony-forming unit, the temperature of said fermentation are 30-36 ℃, and the time of fermentation is 32-48 hour.
16. method according to claim 1, wherein, the raw material of said cellulose is a stalk.
17. according to claim 1 or 16 described methods, wherein, the size of the raw material of said cellulose is 0.2-1 centimetre of a 0.5-3 cm x 0.2-1 cm x.
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