CN101387084B - Method for steam blasting cellulose-containing material - Google Patents
Method for steam blasting cellulose-containing material Download PDFInfo
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- CN101387084B CN101387084B CN2007101215570A CN200710121557A CN101387084B CN 101387084 B CN101387084 B CN 101387084B CN 2007101215570 A CN2007101215570 A CN 2007101215570A CN 200710121557 A CN200710121557 A CN 200710121557A CN 101387084 B CN101387084 B CN 101387084B
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Abstract
The invention provides a steam blasting method for raw materials containing cellulose. The method is to blast the raw materials containing cellulose through steam, wherein the method comprises the following steps of: continuously conveying the raw materials containing cellulose to a heat preservation pressure device, keeping the raw materials containing cellulose in the heat preservation pressure device for 3 to 10 minutes and releasing part of the raw materials containing cellulose in the heat preservation pressure device to atmospheric pressure every other 5 to 12 minutes. The steam blasting method for the raw materials containing cellulose ensures that the raw materials containing cellulose can continuously and fully absorb aqueous vapor under steady conditions, allow the water vapor to rush out during pressure relief to adequately destroy lignin sheathes embedding the cellulose and hemicellulose in the raw materials containing cellulose, so as to increase the proportion of the cellulose which can be in contacted with enzyme in the raw materials containing cellulose, thereby improving the yield of monosaccharide and ethanol prepared from steam blasting products through the method.
Description
Technical field
The invention relates to a kind of preprocess method for preparing the raw material of cellulose, more specifically, is the method about a kind of raw material of steam blasting cellulose-containing.
Background technology
The raw material of cellulose is generally root, stem and the leaf of plant, comprises stalk.Said stalk is remaining, stem and a leaf behind crops (being generally herbaceous plant) results seed or the fruit; Be mainly stem and leaf, as comprise the cereal of maize straw, jowar stalk, rice straw, wheat straw stalk, comprise stalk and the cotton stalk of the oilseed plant of beanstalk.Cellulose, hemicellulose and lignin are combined closely through covalent bond and/or non-covalent bond in the stalk, and the three accounts for the 70-90 weight % of stalk dry gross weight.The content of cellulose, hemicellulose and lignin can the difference to some extent with the different piece of the crops of separate sources or crops in the stalk.
The raw material of said cellulose can be used for the preparation of monose like stalk, and the monose that obtains can be used for fermentative production of ethanol.Cellulose in the raw material of said cellulose can be degraded into glucose under the effect of cellulase; Hemicellulose can be degraded into 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 saccharomyces cerevisiae, pichia stipitis can prepare ethanol by xylose-fermenting.But lignin is a kind of non-polysaccharide material that is polymerized by phenyl-propane, thereby water insoluble being difficult to degrades.Since lignin by the derivative of aromatic hydrocarbon with-C-C-key ,-the O-key is crosslinked in length and breadth; Side chain and hemicellulose are with covalent bonds; Form fine and close network structure---lignin sheath embedding cellulose and hemicellulose; Thereby making cellulase and hemicellulase be difficult for contacting with hemicellulose with cellulose, the result causes stalk also to be difficult to degraded.
Generally, before feedstock production monose and/or ethanol, need carry out preliminary treatment, to destroy the embedding of lignin to cellulose and hemicellulose to the raw material of cellulose by cellulose.The preprocess method of the raw material of the cellulose of prior art generally comprises steam blasting and/or acid hydrolysis.Though acid hydrolysis can destroy lignin, also can destroy 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 blasting, reaches the embedding effect of lignin to cellulose and hemicellulose that destroy.
Document " to the introduction of straw biomass to alcohol conversion process " for example, 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 maize 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, inoranic 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.
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 lignin in the ethanol 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 lignin in the described ethanol extraction steam puffed stalk is in autoclave, adds steam puffed stalk and ethanol, and the volume ratio of steam puffed stalk and ethanol is 1: 10-1: 50, in the pressure limit of 1.5-2.0 MPa, kept 1-2 hour.
Describedly 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 method of using the heat resistant type yeast from the lignin fiber cellulosic biomass, to produce ethanol, and this method may further comprise the steps:
It is the 15-30 millimeter that the lignin fiber cellulosic biomass is ground into granular size; Lignin fiber cellulosic biomass to pulverizing carries out the vapour explosion preliminary 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 isolated by filtration liquid and solid portion, solid portion 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 international units/restrain; Give the suspension of fermentation storage inoculation Marx Crewe Vickers yeast CECT10875 heat resistant type yeast culture; Mixture was shaken under 42 ℃ 72 hours.
But in the preprocess method of the raw material of existing cellulose; The steam blasting preliminary treatment destroys not enough to the lignin sheath of embedding cellulose and hemicellulose in the raw material of cellulose, thereby it is low to cause raw material in the cellulose that adopts this steam blasting to carry out monose and/or ethanol yield that enzymolysis and/or fermenting process of preparing obtain.In addition, the amount of the raw material of the cellulose that existing steam explosion method was handled in the unit interval is less, thereby production efficiency is not high.
Summary of the invention
It is not enough to the lignin sheath destruction of embedding cellulose and hemicellulose in the raw material of cellulose to the objective of the invention is to overcome existing steam blasting preliminary treatment; Cause raw material in the cellulose that adopts this steam blasting to carry out monose and the low shortcoming of ethanol yield that enzymolysis and/or fermenting process of preparing obtain, the method for the raw material of the high steam blasting cellulose-containing of a kind of monose and alcohol yied is provided.
The preprocess method of the raw material of the cellulose of prior art adopts the batch (-type) steam blasting usually; 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 blasting; And insulation a period of time, moment pressure release is then accomplished steam blasting and is collected steam explosion products; Then second batch of fibrous raw material is conveyed into steam blasting device, feeds the pressure of steam to steam blasting once more, and insulation a period of time, moment pressure release is then accomplished steam blasting and is collected steam explosion products; And, according to the method described above subsequently fibrous raw material is carried out the batch (-type) steam blasting according to treating capacity.Inventor of the present invention finds, and is all identical like pressure and temperature even guarantee the condition of each steam blasting 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 blasting 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 blasting condition cause and can't guarantee to make whole cellulose containing raw materials fully to absorb water vapour in the short a few minutes after feeding steam; Therefore; Still the raw material that has most of cellulose be owing to can't fully absorb water vapour, and can not be popped by its inner steam fully, causes the lignin sheath of embedding cellulose and hemicellulose 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 monose and/or ethanol yield.In addition, the method for batch (-type) steam blasting consumed time in the preliminary treatment of raw material is longer, and the treating capacity of raw material is lower in the unit interval, thereby is unfavorable for the raising of production efficiency.
The invention provides a kind of method of raw material of steam blasting cellulose-containing; This method comprises the raw material of steam blasting cellulose-containing; Wherein, This method 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.
The present invention adopts the method for continuous steam blasting that fibrous raw material is carried out preliminary treatment; Can make the raw material of the whole cellulose that place steam blasting device continuous under stable steam blasting condition; In the heat preservation pressure device, fully absorb aqueous vapor; And the inner aqueous vapor of the raw material that makes cellulose changes into water vapour, and steam is gone out by the raw material inside of cellulose when pressure release, fully destroys the lignin sheath of embedding cellulose and hemicellulose in the raw material of cellulose; Increase the cellulose ratios that can contact with enzyme in the raw material of cellulose, thereby improved monose and ethanol yield effectively.And; The method of the raw material of steam blasting cellulose-containing of the present invention is that the continuous raw material that makes cellulose is incubated and keeps pressure in the heat preservation pressure device; The steam blasting of continuous then completion raw material; Therefore, the treating capacity of the raw material of the cellulose that method of the present invention was handled in the unit interval is more, helps the raising of production efficiency.In addition; Under the preferable case; The moisture of raw material that control is used for the cellulose of steam blasting is 30-60 weight %, can further guarantee under the steam blasting condition, to make the inner aqueous vapor of raw material of cellulose to change into water vapour, helps when pressure release steam and is gone out by the raw material inside of cellulose; With the lignin sheath of embedding cellulose and hemicellulose in the raw material of abundant destruction cellulose, thereby further improve monose and ethanol yield.
For example, under all identical situation of other steam blasting condition, the employing moisture of embodiment 1 is that the monose productive rate that the fibrous feedstock production of 40 weight % obtains reaches 43.7%; Alcohol yied reaches 23.9%, and the monose productive rate of embodiment 2 reaches 43.4%, and alcohol yied reaches 23.0%; And the monose productive rate of Comparative Examples 1 is merely 34.3%; Alcohol yied is merely 18.2%, and monose productive rate increase rate is up to 26.5-27.4%, and the alcohol yied increase rate is up to 26.4-31.3%.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 blasting of Comparative Examples 1 of the present invention;
Fig. 3 is the electron scanning micrograph according to stalk behind the method steam blasting of Comparative Examples 2 of the present invention;
Fig. 4 is the electron scanning micrograph according to stalk behind the method steam blasting of the embodiment of the invention 1.
The specific embodiment
The method of the raw material of steam blasting cellulose-containing provided by the invention comprises that the raw material with cellulose is delivered in the heat preservation pressure device continuously; Make the raw material of cellulose in this heat preservation pressure device, keep after 3-10 minute; After being preferably 4-7 minute; 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.
According to the present invention, the raw material of said cellulose is to be delivered to continuously in the heat preservation pressure device, in this device, is continued to transmit, 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 pipeline speed of the raw material of said cellulose is special, and the pipeline 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; Steam is gone out when the pressure release by the raw material inside of cellulose, with the lignin sheath of embedding cellulose and hemicellulose 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 blasting.The condition of said continuous steam blasting can adopt the conventional steam blasting condition of prior art, and like the temperature of steam blasting, promptly the temperature of heat preservation pressure device can be 180-200 ℃, more preferably 185-195 ℃; The pressure of said steam blasting, promptly the pressure of heat preservation pressure device can be 1.4-2.0 MPa, more preferably 1.5-1.6 MPa.Under above-mentioned steam blasting condition, can both fully destroy the lignin sheath of stalk embedding cellulose and hemicellulose, realize the object of the invention.
The method that said raw material with the part cellulose in the heat preservation pressure device is discharged into normal pressure can adopt the method for 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 blasting, 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 blasting of fibrous raw material.
The said amount that at every turn is discharged into the fibrous raw material of normal pressure does not have special qualification; The specification of the heat preservation pressure device that the different production scale is adopted is different; Therefore, the disposal ability of each steam blasting is also different, therefore; Can select the device of different size according to the production needs for use, 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 the cellulose of normal pressure 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 the cellulose of normal pressure 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 transporting velocity of the raw material of cellulose remains unchanged the amount of the raw material of the cellulose in this heat preservation pressure device basically.
In addition; When adopting the raw material of prior art steam blasting cellulose-containing, the raw material that it has been generally acknowledged that said cellulose is maize straw for example, fully water logging; Can obtain the effect of better steam blasting; Because fully water logging can make the raw material of said cellulose for example contain a large amount of water in maize straw inside, when steam blasting, the raw material of the said cellulose for example inner water of maize straw can be gasificated into water vapour; For example go out maize 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 lignin sheath of maize straw embedding cellulose and hemicellulose for example, inventor of the present invention is surprised to find that the moisture of controlling the raw material of the cellulose that is used for steam blasting is 30-60 weight %; Can further guarantee under the steam blasting condition, to make the inner aqueous vapor of raw material of cellulose to change into water vapour; Help when pressure release steam and go out,, thereby further improve monose and ethanol yield with the lignin sheath of embedding cellulose and hemicellulose in the raw material of abundant destruction cellulose by the raw material inside of cellulose.Therefore, under the preferable case, the moisture of the raw material of said cellulose is 30-60 weight %, more preferably 35-50 weight %.
According to the present invention, can obtain the raw material that moisture is the cellulose of 30-60 weight % through variety of way.The raw material of the different batches cellulose that there are differences for moisture can carry out different processing, such as for the high stalk of moisture, can 45-90 ℃ of oven dry part moisture, and make it moisture and be reduced to 30-60 weight %; For the low stalk of moisture, can adopt control water logging time method, the moisture that increases stalk is to 30-60 weight %; Just in time at stalk, can directly carry out steam blasting for moisture to 30-60 weight %.The moisture of the raw material of the preferred said cellulose that is used for steam blasting is 35-50 weight %.Only if specified otherwise; The moisture 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 percentage by weight of the initial weight W1 of cellulose containing raw material, i.e. moisture (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 moisture of the raw material of cellulose remain on 30-60 weight %, is preferably 35-50 weight %, and then is delivered to continuously and carries out steam blasting in the heat preservation pressure device.
The steam explosion products enzymolysis is prepared monose and the step of monose alcohol prepared by fermenting can be passed through this area the whole bag of tricks completion commonly used.Comprise such as, said enzymolysis process and in steam explosion products, to add microbes producing cellulase and/or enzyme that insulation is accomplished under the growth temperature of microbes producing cellulase and/or the great-hearted temperature of enzyme.Said microbes producing cellulase be can eccrine fiber the microbes producing cellulase of plain enzyme.Said enzyme comprises cellulase.
Because growth of microorganism can produce accessory substance; Therefore the preferred enzyme that directly adds, the consumption of said enzyme is The more the better, from cost consideration; Preferably with the dry weight basis of the raw material of every gram cellulose; The consumption of said cellulase is the 8-20 enzyme activity unit, more preferably with the dry weight basis of the raw material of every gram cellulose, and the 10-15 enzyme activity unit.The enzyme activity of cellulase according to the invention is according to American National regenerative resource laboratory (National Renewable 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, and transforming 1 gram Whatman No.1 filter paper in 1 minute is the micrograms of the required enzyme of glucose.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 utilization rate of equipment and installations, and the time of preferred said enzymolysis is 25-48 hour, more preferably 30-40 hour.The pH value of said enzymolysis can be generally 3.0-7.0 for the righttest any action pH of cellulase, and more preferably the pH value is 4.5-5.5.Because the fluctuation of pH value is little in the enzymolysis process; Therefore the pH value of said enzymolysis can be regulated before adding enzyme according to this area method commonly used; For example earlier steam explosion products and water or culture medium (are enzyme-addedly generally mixed with water; Add microbes producing cellulase generally with the culture medium of this microorganism) mix, the solid content that generally makes the gained mixture is 50-70 weight %, according to the pH value of gained mixture; The mixture pH regulator that to treat enzymolysis with sulfuric acid solution or NaOH more preferably is adjusted to the pH value and is 4.5-5.5 to 3.0-7.0.
Said cellulase is a complex enzyme, comprises C at least
1The plain enzyme of fiber type, C
xThe plain enzyme of fiber type and three kinds of enzymes of cellobiase.
C
1Enzyme can make the cellulose of crystallization change amorphous 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 bond 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 bond cuts off β-1 one by one, and the 4-glycosidic bond generates glucose.
Cellobiase then acts on cellobiose, makes the cellobiose enzymolysis generate glucose.
The enzyme that preferred said enzymolysis uses also comprises hemicellulase.Because hemicellulase is degraded into water-soluble wood sugar with hemicellulose; So the enzyme that enzymolysis uses comprises hemicellulase; Exposed fibers is plain more fully on the one hand; Increase the contact probability of cellulose and cellulase, hemicellulose catabolite 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 the raw material of every gram cellulose, 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 per minute 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 reactant liquor 550 nanometers with spectrophotometer.Concrete assay 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 conical flasks, add 16 the gram NaOH after, add 50 ml deionized water; Be heated to the solid matter dissolving with 5 ℃/minute speed water-baths, add 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 sodium azide, obtains the normal wood sugar juice of 1 mg/ml;
Accurately take by weighing 1.000 gram solid hemicellulases or pipette 1 milliliters of liquid hemicellulase stoste; 0.1 mol acetate-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 water-bath heating 2 milliliters of wood sugar gradient standard liquids (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 standard liquid is mixed with the normal wood sugar juice 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 standard liquid after distinguishing assaying reaction under 550 nanometers; With the absorbance value is abscissa, and xylose concentration is an ordinate drawing standard curve.Can get regression equation y=bx+a by this calibration curve, wherein, x is an absorbance value, and y is an xylose concentration, and a is the intercept of gained linear equation, and b is the slope of gained linear 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 standard liquid.And according to the vigor of computes hemicellulase:
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 said method, and then calculate the consumption of hemicellulase.
Monose in the enzymolysis product can be through sugar industry method separation and purification commonly used.For example remove the solid matter (raw material of the cellulose of for example not degraded) in the enzymolysis product through filtration, centrifugation method.Collect filtrating or supernatant, evaporation is removed part moisture and is obtained the supersaturation syrup that contents of monosaccharides is 60-65 weight %.In the vacuum crystallization jar, through transpiring moisture the monose crystallization is separated out, crystal and syrup form massecuite; When the crystal volume meets the requirements, massecuite is transferred to crystallizer from the vacuum crystallization jar, cooling is further separated out uncrystallized monose gradually.Carry out Separation of Solid and Liquid with centrifuge, crystallization monose and molasses get product.Molasses are repeated said process crystallization 2-4 time, can fully extract monose.Crystallization process can adopt nature graining method, stimulate graining method and nucleus graining method graining, preferably in syrup, adds crystallization with the nucleus graining method graining of kind of paste as crystal seed.
Step by the monose alcohol prepared by fermenting can be accomplished through this area the whole bag of tricks commonly used; For example; Can ferment pentoses and/or the microorganism of hexose may be used to process by the monose alcohol prepared by fermenting because saccharomyces cerevisiae is the microorganism of the zymohexose that ethanol-tolerant, accessory substance are few, alcohol yied is high of widespread usage on the brewing industry; Pichia stipitis for both can ferment pentoses also can zymohexose microorganism (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 saccharomyces cerevisiae.In every gram enzymolysis product, the inoculum concentration of the employed yeast of said fermentation is 10
3-10
8CFU, more preferably 10
4-10
6CFU.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 brewer's yeast of ATCC numbering 2601).The CFU of said yeast 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 light microscopes,, can get the number of viable bacteria in 1 gram dry ferment 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 ferment, also can or be purchased the activated seed liquid of bacterial classification for dry ferment.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 the purity of ethanol 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 said steam blasting 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 during the fermentation from influencing the growth of yeast, and preferably before enzymolysis, adding bacteria antibiotic such as industrial penicillin, said antibiotic not have effect to yeast but the growth of the bacterium that can suppress to mix.With every milliliter of enzymolysis liquid or zymotic fluid is benchmark, and said antibiotic addition is a 1-10 unit.Said enzymolysis liquid comprises steam explosion products, enzyme and water, and said zymotic fluid comprises the yeast of enzymolysis product and inoculation.
Because under the high-temperature and high-pressure conditions of steam blasting; The raw material of cellulose can produce the mortifier such as furfural, furans; Said mortifier 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 centrifugation steam explosion products then owing to above-mentioned mortifier is prone to be dissolved in the water, thereby can be removed mortifier through washing from steam explosion products.The temperature of said washing is high more, and the mortifier that can dissolve is many more, considers energy consumption, and the temperature of preferred said washing is 60-90 ℃, more preferably 60-80 ℃.Because the water that washing is added is many more; The mortifier that then dissolves is also many more; But consider the energy consumption of centrifugation, preferably with the dry weight basis of the said cellulose of every gram, the amount of water of said washing is the 2-10 gram; More preferably with the dry weight basis of the raw material of every gram cellulose, the amount of water of said washing is the 2-3 gram.
Owing to may contain sandstone impurity and iron tramp in the raw material of cellulose such as the maize straw; Can cause damage to steam blasting equipment; Therefore adopting steam explosion products of the present invention to prepare the method for monose and/or ethanol; Can comprise the existing stone deironing routine operation that removes, 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 blasting equipment by wind because quality is big, iron tramp since the attraction of magnet can not get in the steam blasting equipment with raw material yet, thereby can accomplish except that the stone deironing.In addition; Because the raw material of cellulose itself is entangled with and the occluding device pipeline easily; Therefore the size of the raw material of preferred said cellulose is 0.2-1 centimetre of a 0.5-3 cm x 0.2-1 cm x, and more preferably the size of 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 explained method and the method that is prepared monose and ethanol by this steam explosion products of the raw material of steam blasting cellulose-containing of the present invention.
(1) continuous steam blasting
Test maize straw raw material as shown in Figure 1; The moisture that records maize 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 moisture is that the maize 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 moisture of this maize straw section be 40 weight %.Be that the maize straw of 40 weight % is delivered in the heat preservation pressure device with 2000 these moisture of gram continuously then, the pressure of said heat preservation pressure device is 1.6 MPas, and temperature is 195 ℃; Said maize straw transmits in this heat preservation pressure device continuously, and the linear velocity of transmission makes this maize 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 closing; Make the part maize straw be discharged into the normal pressure ejection, accomplish steam blasting, and repeat said process and carry out continuous steam blasting.The raw material that at every turn is discharged into the cellulose of normal pressure 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 Separation of Solid and Liquid then, obtain 3000 gram solid steam explosion products (moisture is 60 weight %) altogether.
The mensuration of cellulose gross weight and hemicellulose 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 2.0 grams, 300.0 milligrams of this dried steam explosion products of weighing are positioned in heavy 80 100 milliliters of dry conical flasks restraining.In said conical flask, adding 3.00 ml concns is the sulfuric acid solution of 72 weight %, stirs 1 minute.Then conical flask was heated 60 minutes in 30 ℃ of water-baths, every at a distance from 5 minutes with stirring once to guarantee even hydrolysis.After water-bath finished, the concentration dilution that makes sulfuric acid with deionized water was to 4 weight %.Filter the steam explosion products acid hydrolysis liquid after the said dilution with Buchner funnel, obtain altogether filtrating 84 milliliters.20 milliliters of filtratings are transferred in the triangular flask of 50 milliliters of dryings.Use 2.5 gram calcium carbonate to regulate this pH value of filtrate to 5.5, left standstill 5 hours, collect supernatant liquor.0.2 the supernatant liquor that the micron membrane filtration is collected, the filtrating of gained supernatant liquor is analyzed with Biorad Aminex HPX-87P high performance liquid chromatography (HPLC).HPLC condition: sample size 20 microlitres; Flowing phase is 0.2 micron membrane filtration, and the HPLC ultra-pure 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 running 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 sample.HPLC analyzes and to obtain that concentration of glucose is 2.22 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 0.186 gram; Because being the sulfuric acid solution of 72 weight %, concentration can the cellulose of the product of steam blasting all be hydrolyzed into glucose; Therefore the weight of gained glucose is 1.11 times of cellulosic weight in the steam explosion products; Cellulose 0.168 gram in the steam explosion products after the i.e. said washing of 1 gram then is total to cellulose 504 grams in the 3000 gram steam explosion products.HPLC analyzes and to obtain that xylose concentration is 0.54 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 0.055 gram; Because being the sulfuric acid solution of 72 weight %, concentration can hemicellulose in the product of steam blasting all be hydrolyzed into wood sugar; Therefore the weight of gained wood sugar is 1.14 times of weight of hemicellulose in the steam explosion products; Promptly contain hemicellulose 0.048 gram in the 1 gram steam explosion products, then contain hemicellulose 144 grams altogether in the 3000 gram steam explosion products.
(2) enzymolysis
With remaining washing steam explosion products behind step (1) sampling and testing, mix with 1000 gram water, regulate pH value to 5.0; After being heated to 50 ℃; With the dry weight basis of every gram stalk, add the cellulase (jade of the He family Bioisystech Co., Ltd) of 20 enzyme activity units, and be incubated 40 hours down at 50 ℃.Enzymolysis product is filtered with Buchner funnel, 20 milliliters of filtratings are transferred in 50 milliliters of dry triangular flasks, left standstill 5 hours, collect supernatant liquor.0.2 the supernatant liquor that the micron membrane filtration is collected, according to the said efficient liquid-phase condition of step (1), the glucose of measuring and calculating in the enzymolysis product 524.8 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 472.8 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, result of calculation is seen table 1.
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 every gram enzymolysis product, inoculation is 10
5The saccharomyces cerevisiae of CFU (the super highly active dry yeast in Angel, Hubei Angel Yeast joint-stock company), gained mixture under 35 ℃ in fermentation tank stir culture 40 hours.At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 286.8 grams at 78.3 ℃ of following second distillations, and according to the computes alcohol yied, result of calculation is seen table 1.
Alcohol yied=100% * ethanol weight/stalk dry weight
Embodiment 2
Present embodiment is explained method and the method that is prepared monose and ethanol by this steam explosion products of the raw material of steam blasting cellulose-containing of the present invention.
Method according to embodiment 1 prepares ethanol; Different is; Maize straw section with moisture 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 moisture of this maize straw section.Be that the maize straw of 75 weight % is delivered to and carries out continuous steam blasting in the heat preservation pressure device with 2000 these moisture of gram continuously according to the condition of embodiment 1 and method then, obtain 3000 gram solid steam explosion products (moisture is 83.43 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, hemicellulose 60 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 216.8 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 195.3 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Second distillation can get ethanol 114.9 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, result of calculation is seen table 1.
Embodiment 3
Present embodiment is explained method and the method that is prepared monose and ethanol by this steam explosion products of the raw material of steam blasting cellulose-containing of the present invention.
Method according to embodiment 1 prepares ethanol; Different is; Be not carry out water after the maize straw section of 20 weight % is sent into continuously to drench directly to be delivered to and carry out continuous steam blasting in the heat preservation pressure device from feeder with 2000 gram moisture; The condition of steam blasting is collected steam explosion products and is also obtained 3556 gram solid steam explosion products (moisture 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 3553 gram steam explosion products cellulose 672 grams altogether, hemicellulose 196.8 grams.
Method according to embodiment 1 is carried out enzymolysis and fermentation to above-mentioned steam explosion products, and according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 690.0 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 621.6 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11.Second distillation can get ethanol 362.3 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, result of calculation is seen table 1.
Comparative Examples 1
The method of the raw material of the steam blasting cellulose-containing of this Comparative Examples explanation prior art and the method for preparing monose and ethanol by this steam explosion products.
Method according to embodiment 1 prepares ethanol, and different is that (Zhang Xuefeng, orchid are kindly helped secure the success of, wine brewing according to document " to the introduction of straw biomass to alcohol conversion process "; The 29th the 4th phase of volume; The 37-38 page or leaf, in July, 2002) disclosed method, stalk was soaked in water earlier before steam blasting 40 minutes; The moisture that records the maize straw raw material after the immersion reaches 83 weight %; The maize straw of getting 2000 these moisture of gram then and be 83 weight % (every batch 1000 gram) in two batches carries out the batch (-type) steam blasting, and the method for said batch (-type) steam blasting is for keeping 5 minutes pressure release then with the said maize straw of 1000 grams earlier with the pressure of 1.6 MPas under 195 ℃; And then the said maize straw of remaining 1000 grams carried out steam blasting under same condition, obtain 1133 gram steam explosion products 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 1133 gram steam explosion products cellulose 142.8 grams altogether, hemicellulose 51 grams.
Carry out enzymolysis and fermentation according to the method for embodiment 1 then, and according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 117.3 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 105.7 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11; Second distillation obtains ethanol 62 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, result of calculation is seen table 1.
Comparative Examples 2
The method of the raw material of the steam blasting cellulose-containing of this Comparative Examples explanation prior art and the method for preparing monose and ethanol by this steam explosion products.
Method according to embodiment 1 prepares ethanol; Different is; With 2000 gram moisture is that 20 weight % maize straws were directly kept under the pressure with 1.8 MPas under 190 ℃ 4 minutes; Steam blasting is accomplished in pressure release then, collects steam explosion products and also obtains 4000 gram steam explosion products (moisture 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 4000 gram steam explosion products cellulose 680 grams altogether, hemicellulose 224 grams.
Carry out enzymolysis and fermentation according to the method for embodiment 1 then, and according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 579.7 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 522.2 is restrained by the cellulosic weight of enzymolysis in the steam explosion products divided by 1.11; Second distillation obtains ethanol 302.4 grams.And calculate cellulose conversion rate, monose productive rate and alcohol yied according to the method for embodiment 1 and formula, result of calculation 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 vapour, so steam blasting can not fully destroy stalk from inside to outside; As can beappreciated from fig. 3, the effect of Comparative Examples 1 maize straw steam blasting and bad, cellulose still is pencil, and cellulose surface is residual that the lignin sheath arranged; And the steam blasting effect of embodiment 1 as shown in Figure 4 is fine, can see free cellulose.
Embodiment 4
Present embodiment is explained method and the method that is prepared monose and ethanol by this steam explosion products of the raw material of steam blasting cellulose-containing of the present invention.
(1) continuous steam blasting
Method according to embodiment 1 is carried out continuous steam blasting to maize straw; Different is; Water drenches that to make the moisture that gets into the maize straw before the heat preservation pressure device be 65 weight %, and the maize straw that is 65 weight % with this this moisture of 1000 gram continuously is delivered to and carries out continuous steam blasting in the heat preservation pressure device.The raw material that at every turn is discharged into the cellulose of normal pressure 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 Separation of Solid and Liquid then, obtain 1300 gram solid steam explosion products (moisture is 73.1 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 1300 gram steam explosion products cellulose 150.5 grams altogether, hemicellulose 45.5 grams.
(2) enzymolysis
With remaining steam explosion products behind step (1) sampling and testing, mixes with 400 gram water, regulate pH value to 6.0, be heated to 55 ℃ after, with the dry weight basis of every gram stalk, add the cellulase of 15 enzyme activity units, and descend to be incubated 30 hours at 55 ℃.Enzymolysis product is filtered with Buchner funnel, 20 milliliters of filtratings are transferred in the triangular flask of 50 milliliters of dryings, left standstill 5 hours, collect supernatant liquor.0.2 the supernatant liquor that the micron membrane filtration is collected, according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 154.8 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 139.5 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, result of calculation is seen table 1.
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 32 ℃; In every gram enzymolysis product; Inoculation is the saccharomyces cerevisiae (the super highly active dry yeast in Angel, Hubei Angel Yeast joint-stock company) of 106 CFUs, the gained mixture under 32 ℃ in fermentation tank stir culture 35 hours.At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 80.5 grams at 78.3 ℃ of following second distillations, and according to the computes alcohol yied, result of calculation is seen table 1.
Alcohol yied=100% * ethanol weight/stalk dry weight
Embodiment 5
Present embodiment is explained method and the method that is prepared monose and ethanol by this steam explosion products of the raw material of steam blasting cellulose-containing of the present invention.
(1) continuous steam blasting
Method according to embodiment 1 is carried out continuous steam blasting to maize straw; Different is; Be after the maize straw of 20 weight % is cut into the segment of 0.5 centimetre of 2.5 cm x, 0.5 cm x earlier with moisture; With vertical macerator (the auspicious mechanical Co., Ltd in river in Shangdong Province, Shandong) and weight is after 1/3rd water of said maize straw raw material weight mixes, and can get the maize straw of moisture 50 weight %.The maize straw that is 50 weight % with this 1000 gram moisture 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 maize straw transmits in this heat preservation pressure device continuously, and the linear velocity of transmission makes this maize 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 closing; Make the part maize straw be discharged into the normal pressure ejection, accomplish steam blasting, and repeat said process and carry out continuous steam blasting.The raw material that at every turn is discharged into the cellulose of normal pressure 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 blasting, collect not totally 1250 gram steam explosion products (moisture is 60 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 1250 gram steam explosion products cellulose 215 grams altogether, hemicellulose 62.5 grams.
(2) enzymolysis
With remaining steam explosion products behind step (1) sampling and testing, mixes with 400 gram water, regulate pH value to 6.0, be heated to 55 ℃ after, with the dry weight basis of every gram stalk, add the cellulase of 15 enzyme activity units, and descend to be incubated 30 hours at 55 ℃.Enzymolysis product is filtered with Buchner funnel, 20 milliliters of filtratings are transferred in the triangular flask of 50 milliliters of dryings, left standstill 5 hours, collect supernatant liquor.0.2 the supernatant liquor that the micron membrane filtration is collected, according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 212.4 restrains totally.The glucose weight that said enzymolysis obtains promptly totally 191.4 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, result of calculation is seen table 1.
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 32 ℃, in every gram enzymolysis product, inoculation is 10
6The saccharomyces cerevisiae of CFU (the super highly active dry yeast in Angel, Hubei Angel Yeast joint-stock company), gained mixture under 32 ℃ in fermentation tank stir culture 35 hours.At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 112.6 grams at 78.3 ℃ of following second distillations, and according to the computes alcohol yied, result of calculation is seen table 1.
Alcohol yied=100% * ethanol weight/stalk dry weight
Embodiment 6
Present embodiment is explained method and the method that is prepared monose and ethanol by this steam explosion products of the raw material of steam blasting cellulose-containing of the present invention.
(1) continuous steam blasting
Getting 1000 gram moisture is the maize straw of 35 weight %, is cut into the segment of 0.7 centimetre of 1.2 cm x, 0.4 cm x, and the maize straw that is 35 weight % with this 1000 gram moisture 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 maize straw transmits in this heat preservation pressure device continuously; The linear velocity that transmits makes this maize 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 closing makes the part maize straw be discharged into the normal pressure ejection; Accomplish steam blasting, and repeat said process and carry out continuous steam blasting.The raw material that at every turn is discharged into the cellulose of normal pressure 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 centrifugal pump LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out Separation of Solid and Liquid then, obtain 2600 gram steam explosion products (moisture 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 2600 gram steam explosion products cellulose 286 grams altogether, hemicellulose 84.5 grams.
(2) enzymolysis
With remaining washing steam explosion products behind step (1) sampling and testing; Mix with 1000 gram water; Regulate pH value to 4.0, be heated to 45 ℃ after, with the dry weight basis of every gram stalk; Add the cellulase of 18 enzyme activity units and the hemicellulase of 8.0 enzyme activity units (Beijing chemical reagents corporation), and be incubated 35 hours down at 45 ℃.Enzymolysis product is filtered with Buchner funnel, 20 milliliters of filtratings are transferred in the triangular flask of 50 milliliters of dryings, left standstill 5 hours, collect supernatant liquor.0.2 the supernatant liquor that the micron membrane filtration is collected, according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 291.7 restrains totally, and wood sugar is totally 68.4 grams.The glucose weight that said enzymolysis obtains is divided by 1.11; Be totally 262.8 to be restrained by the cellulosic weight of enzymolysis in the steam explosion products; The wood sugar weight that said enzymolysis obtains is divided by 1.14; Be totally 60 to be restrained by the weight of the hemicellulose of enzymolysis in the steam explosion products, according to computes cellulose conversion rate, hemicellulose conversion ratio and monose productive rate, result of calculation is seen table 1.
Cellulose conversion rate=100% * by the cellulosic weight of enzymolysis/cellulosic gross weight
Hemicellulose conversion ratio=100% * by the gross weight of the weight/hemicellulose of the hemicellulose 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 33 ℃, in every gram enzymolysis product, inoculation is 10
5The saccharomyces cerevisiae of CFU (ATCC numbering 2601) and 10
3The pichia stipitis of CFU (ATCC numbering 58376), under 33 ℃ in fermentation tank stir culture 39 hours.At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 187.3 grams at 78.3 ℃ of following second distillations, and according to the computes alcohol yied, result of calculation is seen table 1.
Alcohol yied=100% * ethanol weight/stalk dry weight
Embodiment 7
Present embodiment is explained method and the method that is prepared monose and ethanol by this steam explosion products of the raw material of steam blasting cellulose-containing of the present invention.
(1) continuous steam blasting
Getting 1000 gram moisture is the maize straw of 55 weight %, is cut into the segment of 0.8 centimetre of 2.0 cm x, 0.6 cm x, and the maize straw that is 55 weight % with this 1000 gram moisture 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 maize straw transmits in this heat preservation pressure device continuously; The linear velocity that transmits makes this maize 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 closing makes the part maize straw be discharged into the normal pressure ejection; Accomplish steam blasting, and repeat said process and carry out continuous steam blasting.The raw material that at every turn is discharged into the cellulose of normal pressure 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 centrifugal pump LW400 type horizontal screw centrifuge (Zhangjiagang Huada Centrifugal Manufacturing Co., Ltd.) under 900 rev/mins of rotating speeds, to carry out Separation of Solid and Liquid then, obtain 1500 gram steam explosion products (moisture is 70 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 1500 gram steam explosion products cellulose 195.8 grams altogether, hemicellulose 57.6 grams.
(2) enzymolysis
With remaining washing steam explosion products behind step (1) sampling and testing; Mix with 800 gram water; Regulate pH value to 3.5, be heated to 50 ℃ after, with the dry weight basis of every gram stalk; Add the cellulase of 10 enzyme activity units and the hemicellulase of 4.5 enzyme activity units (Beijing chemical reagents corporation), and be incubated 48 hours down at 50 ℃.Enzymolysis product is filtered with Buchner funnel, 20 milliliters of filtratings are transferred in the triangular flask of 50 milliliters of dryings, left standstill 5 hours, collect supernatant liquor.0.2 the supernatant liquor that the micron membrane filtration is collected, according to the said efficient liquid-phase condition of embodiment 1 step (1), the glucose of measuring and calculating in the enzymolysis product 198.2 restrains totally, and wood sugar is totally 46 grams.The glucose weight that said enzymolysis obtains is divided by 1.11; Be totally 178.6 to be restrained by the cellulosic weight of enzymolysis in the steam explosion products; The wood sugar weight that said enzymolysis obtains is divided by 1.14; Be totally 40.3 to be restrained by the weight of the hemicellulose of enzymolysis in the steam explosion products, according to computes cellulose conversion rate, hemicellulose conversion ratio and monose productive rate, result of calculation is seen table 1.
Cellulose conversion rate=100% * by the cellulosic weight of enzymolysis/cellulosic gross weight
Hemicellulose conversion ratio=100% * by the gross weight of the weight/hemicellulose of the hemicellulose 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 every gram enzymolysis product, inoculation is 10
4The saccharomyces cerevisiae of CFU and 10
4The pichia stipitis of CFU, under 34 ℃ in fermentation tank stir culture 37 hours.At 100 ℃ of distillation gained tunnings, the gained distillation fraction can get ethanol 127 grams at 78.3 ℃ of following second distillations, and according to the computes alcohol yied, result of calculation is seen table 1.
Alcohol yied=100% * ethanol weight/stalk dry weight
Table 1
| Embodiment or Comparative Examples | Embodiment 1 | Comparative Examples 1 | Comparative Examples 2 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 | Embodiment 6 | Embodiment 7 |
| Cellulose conversion rate (%) | 93.8 | 74.0 | 76.8 | 93.0 | 92.5 | 92.7 | 89.0 | 91.9 | 91.2 |
| Hemicellulose conversion ratio (%) | - | - | - | - | - | - | - | 71.0 | 70.0 |
| Monose productive rate (%) | 43.7 | 34.3 | 35.9 | 43.4 | 43.1 | 44.2 | 42.5 | 55.4 | 54.3 |
| Alcohol yied (%) | 23.9 | 18.2 | 19.0 | 23.0 | 22.6 | 23.0 | 22.5 | 28.8 | 28.2 |
Can find out from table 1; Employing is prepared in the method for monose and ethanol by the steam explosion products that the method for the fibrous raw material of steam blasting of the present invention obtains, and cellulose conversion rate, monose productive rate and alcohol yied all are significantly improved than the method for Comparative Examples 1 and Comparative Examples 2.
Claims (7)
1. the method for the raw material of a steam blasting cellulose-containing; This method comprises the raw material of steam blasting cellulose-containing; It is characterized in that; This method 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, and the raw material that at every turn is discharged into the cellulose of normal pressure is the 6-15 weight % of the raw material of cellulose in the heat preservation pressure device; The moisture of the raw material of said cellulose is 30-60 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 the cellulose of normal pressure 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, the moisture of the raw material of said cellulose is 35-50 weight %.
6. according to any described method in the claim 1,2,3 or 5, 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.
7. method according to claim 1, wherein, the raw material of said cellulose is a stalk.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007101215570A CN101387084B (en) | 2007-09-10 | 2007-09-10 | Method for steam blasting cellulose-containing material |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101988074B (en) * | 2009-07-31 | 2012-11-14 | 中粮集团有限公司 | Enzymolysis method |
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| CN101387084B (en) * | 2007-09-10 | 2012-05-30 | 中粮集团有限公司 | Method for steam blasting cellulose-containing material |
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| CN114376073B (en) * | 2022-02-09 | 2022-09-30 | 紫茎泽兰中医农业科技(云南)有限公司 | Non-grain type monogastric animal feed and preparation method thereof |
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| CN101387084A (en) * | 2007-09-10 | 2009-03-18 | 中粮集团有限公司 | Method for steam blasting cellulose-containing material |
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| CN101387084A (en) * | 2007-09-10 | 2009-03-18 | 中粮集团有限公司 | Method for steam blasting cellulose-containing material |
| CN201200867Y (en) * | 2008-05-19 | 2009-03-04 | 河南省泰隆科技开发应用有限公司 | Continuous exploding spray pulping device |
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| CN101988074B (en) * | 2009-07-31 | 2012-11-14 | 中粮集团有限公司 | Enzymolysis method |
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Application publication date: 20090318 Assignee: COFCO Nutrition and Health Research Institute Co., Ltd. Assignor: Cofco Group Co., Ltd. Contract record no.: 2012990000627 Denomination of invention: Method for steam blasting cellulose-containing material Granted publication date: 20120530 License type: Exclusive License Record date: 20120827 |