CN102839198B - Method for enhancing alkaline hydrogen peroxide to pretreat lignocellulose by ultrasonic wave - Google Patents
Method for enhancing alkaline hydrogen peroxide to pretreat lignocellulose by ultrasonic wave Download PDFInfo
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- CN102839198B CN102839198B CN201210339024.0A CN201210339024A CN102839198B CN 102839198 B CN102839198 B CN 102839198B CN 201210339024 A CN201210339024 A CN 201210339024A CN 102839198 B CN102839198 B CN 102839198B
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
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Abstract
The invention relates to a method for enhancing alkaline hydrogen peroxide to pretreat lignocellulose by an ultrasonic wave, and the method comprises the steps that when the lignocellulose is subjected to alkali treatment, the enhancing effect of the ultrasonic wave is conducted; after a period of time, the hydrogen peroxide is added for strong oxidization; and after the pretreatment is finished, a raw material is subjected to solid-liquid separation, wherein after a solid component in the raw material is dried, cellulase enzymolysis and composition analysis are conducted, and a liquid component is used for recovering hemicellulose and lignin components. The method aims at enhancing saponification and the strong oxidization of the alkaline hydrogen peroxide on the lignin component by fierce cavitation of the ultrasonic wave, so that the content of the lignin hindering the enzymolysis of a lignocellulose raw material is reduced greatly, most cellulose and hemicellulose are degraded to be fermentable monosaccharide, and the raw material utilization ratio of the lignocellulose is increased to the greatest extent. In addition, the hemicellulose and the lignin in pretreatment liquid are recovered by an acidulation method, and the green process without wasting of resources is realized.
Description
Technical field
The invention belongs to the biomass recycling use technical field, relate to the intensified by ultrasonic wave alkaline peroxide impregnation, particularly, utilize hyperacoustic cavatition to strengthen saponification and the strong oxidation of alkaline hydrogen peroxide to the xylogen composition, with the decrease content of lignin, shorten action time, improve enzymolysis efficiency.
Background technology
The s-generation alcohol fuel that the lignocellulose of take is raw material can be alleviated increasingly serious energy shortage and problem of environmental pollution on the one hand, can also reduce on the other hand the waste of lignocellulosic material, realizes recycling.Yet, how to improve in process of production the percent hydrolysis of lignocellulosic material and the bottleneck that utilization ratio remains the development of restriction s-generation Fuel Ethanol.
Lignocellulosic material three major polymers: cellulose, hemicellulose and xylogen form, and wherein, Mierocrystalline cellulose is to be connect the long chain molecule formed by β-Isosorbide-5-Nitrae glycosidic link by D-Glucose, and long chain molecule further forms a kind of rock steady structure with highly crystalline district.And Mierocrystalline cellulose is inlayed by hemicellulose and xylogen parcel, mutually be wound around each other and form complexity structure closely, be difficult to make directly degraded cellulose components wherein of cellulase, therefore need to remove xylogen wherein by effective pretreatment mode, destroy the dense structure of lignocellulose, reduce degree of crystallinity, increase specific surface area, this is also committed step and the technology prerequisite of successfully carrying out converting fuel ethanol.
The more pretreatment process of application has Physical as mechanical disintegration, microwave treatment, ultrasonication etc. at present, and chemical method is as acid treatment, alkaline purification, wet oxidation processing, ozonize etc., and physico-chemical processes is processed and biological process as steam pop.But traditional single pretreatment process all is difficult to the xylogen in lignocellulosic material is reduced to minimum level, do not give abundant attention to the lignin recovery utilization of removing, and application sourer pre-treatment all can produce the fermentation inhibitors such as furfural, HMF.Therefore should improve the enzymatic hydrolyzation of raw material based on reducing to greatest extent the xylogen composition that hinders cellulase hydrolysis, reduce the generation of fermentation inhibitor, and the purpose that the xylogen composition is recycled, suitable pretreatment mode found.Consider different pretreatments advantage separately, it can be carried out to the advantage combination, explore the pretreated new approaches of lignocellulose, break through the bottleneck of conventional pretreatment mode.
Summary of the invention
Technical problem: the object of the present invention is to provide a kind of method of intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose, improve enzymolysis yield and the raw material availability of lignocellulose, overcome the defect that the conventional pretreatment technology exists.It is the alkaline peroxide impregnation of intensified by ultrasonic wave, cavatition that can the combining ultrasonic ripple and saponification, the strong oxidation of alkaline hydrogen peroxide, remove the xylogen composition that suppresses Mierocrystalline cellulose and hemicellulose enzymolysis, improve the enzymatic hydrolyzation of enzymolysis raw material, be conducive to follow-up fermenting process, shorten action time, and realize the wherein recovery of composition.
Technical scheme: the method for intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose of the present invention mainly comprises the operations such as lignocellulose pre-treatment, the separation of solid-liquid component, component recovery, composition analysis, cellulase hydrolysis, is specifically related to step as described below:
1) air-dry lignocellulose being carried out to the physical pulverization processing sieves again;
2) lignocellulose after screening is dried to constant weight in 105 ℃, remove wherein moisture;
3) lignocellulose after drying is added in sodium hydroxide solution and is processed, and carry out ul-trasonic irradiation simultaneously, after the 1～50min of interval, add hydrogen peroxide to continue to process, obtain pretreated reaction paste,
4) pretreated reaction paste in step 3) is carried out to solid-liquid separation, collect liquid material, constantly wash solid substances to pH value and be neutral and it is dried to constant weight under 105 ℃;
5) liquid material of collecting in step 4) is carried out to the recovery of hemicellulose and xylogen: utilize HCl solution that the pH value of liquid material is adjusted to 5～6, add the ethanol of 3 times of volumes, filter after standing 2h, filter residue is hemicellulose; Get its filtrate rotary evaporation that reduces pressure concentrated under 90 ℃, continue to drip HCl solution to pH value 1.5～2 after ethanol steams fully, the standing Precipitation for the treatment of, centrifugal rear gained solid is xylogen;
6) in the citric acid-sodium citrate damping fluid that is 4.5～4.8 by the solid substances after drying in step 4) in the pH value, with cellulase, be hydrolyzed;
7) slurry obtained after hydrolysis in step 6) is carried out to solid-liquid separation, liquid material is mainly the hydrolysis sugars such as glucose, and solid substances is the part hard-degraded substance.
The described screening particle diameter of step 1) is 20 orders～60 orders.
The described NaOH strength of solution of step 3) is 0.125～0.5mol/L, H
2o
2volume fraction be 20～40%.
The solid-to-liquid ratio of the described lignocellulose of step 3) and NaOH solution is 1:5～1:15g/mL, lignocellulose and H
2o
2solid-to-liquid ratio be 1:0.1～1:1g/mL.
The described ultrasonic wave of step 3) and coefficient total time of NaOH solution are 10～60min, and hyperacoustic operating power is 80～300W, and temperature is 25～80 ℃.
The described cellulase heap(ed) capacity of step 6) is the dry raw material of 5～50FPU/g, and the solid-to-liquid ratio of dry raw material and citric acid-sodium citrate buffer is 1:10～1:100g/mL, and hydrolysis temperature is 50 ℃, and enzymolysis time is 12～72h.
Beneficial effect: with the pre existing treatment technology, compare, the method for intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose provided by the invention has the following advantages:
1) remove most xylogen compositions, reduce its inhibition to the lignocellulosic material enzymolysis, improve dhdps enzyme solution yield;
2) do not produce the fermentation inhibitors such as furfural, HMF in preprocessing process, be conducive to improve the alcohol yied in follow-up fermenting process;
3) pre-treatment operates at normal temperatures and pressures, and action time is short, and mild condition is easy to control, energy efficient and cost of investment;
4) by the xylogen of removal and part hemicellulose, the mode with acid out is reclaimed, for the complete utilization of biomass resource provides basis.
The accompanying drawing explanation
Fig. 1 utilizes ultrasonic wave and alkaline hydrogen peroxide to realize the process flow sheet of lignocellulose pre-treatment and efficient enzymolysis.
Fig. 2 adopts the present invention to process the composition analysis comparison diagram of corn cob in different instances.
Specific embodiments
Enzymolysis yield method of calculation after Raw enzymolysis of the present invention are:
Mierocrystalline cellulose and hemicellulose total amount in enzymolysis yield (%)=(glucose total amount * 0.9 * 100)/enzymolysis substrate.
Comparative Examples 1---pretreatment mode: took 40 mesh sieves and dried rear corn cob 10g, and added 100ml, NaOH solution and the 3.3ml30%(v/v of 0.25mol/L) H
2o
2, temperature is 25 ℃, reaction 24h.The same embodiment of enzymolysis process, the enzymolysis yield is 81.05%.
Comparative Examples 2---pretreatment mode: take 40 mesh sieves and dried rear corn cob 10g, added 100ml, the NaOH solution of 0.25mol/L, temperature is 25 ℃, after reaction 12h, adds 3.3ml30%(v/v) H
2o
2continue reaction 12h.The same embodiment of enzymolysis process, the enzymolysis yield is 75.23%.
Embodiment 1
Took 40 mesh sieves and dried rear corn cob 10g, added 100ml, NaOH solution and the 3.3ml30%(v/v of 0.25mol/L) H
2o
2be placed in beaker, and carry out ul-trasonic irradiation simultaneously, hyperacoustic power that applies is 120W, and operative temperature is 25 ℃, and ultrasonic total time is 40min.Reaction is carried out vacuum filtration by slurry after finishing, and realizes solid-liquid separation, and the filter residue of above-mentioned generation is carried out to the flushing of distill repeatedly water until the pH value is neutral, and dries to constant weight in 105 ℃.
The corn cob that pre-treatment is obtained carries out normal form analytical method (VanSoest) and measures (main component as shown in Figure 2) and enzymolysis, solid-to-liquid ratio 1:70(g/mL), the time is that 72h, temperature are that citric acid-sodium citrate buffer, the cellulase that 50 ℃, pH are 4.8 produced by Trichodermareesei wherein enzymatic hydrolysis condition is:, its heap(ed) capacity is the dry raw material of 20FPU/g.After enzymolysis finishes, reacting slurry is taken out, after the enzyme that goes out 10min alive, carry out vacuum filtration in boiling water bath.Reducing sugar composition in filtrate is measured by the DNS method, and the enzymolysis yield is 72.1%.
Embodiment 2
Take 40 mesh sieves and dried rear corn cob 10g, add 100ml, 0.25mol/L NaOH solution be placed in beaker, and carry out ul-trasonic irradiation simultaneously, hyperacoustic power that applies is 120W, operative temperature is 25 ℃, and ultrasonic total time is 40min, after ul-trasonic irradiation 10min, adds and 3.3ml30%(v/v) H
2o
2.Reaction is carried out vacuum filtration by slurry after finishing, and realizes solid-liquid separation, and the filter residue of above-mentioned generation is carried out to the flushing of distill repeatedly water until the pH value is neutral, and dries to constant weight in 105 ℃.
The corn cob that pre-treatment is obtained carries out normal form analytical method (VanSoest) and measures (main component as shown in Figure 2) and enzymolysis, solid-to-liquid ratio 1:70(g/mL), the time is that 72h, temperature are that citric acid-sodium citrate buffer, the cellulase that 50 ℃, pH are 4.8 produced by Trichodermareesei wherein enzymatic hydrolysis condition is:, its heap(ed) capacity is the dry raw material of 20FPU/g.After enzymolysis finishes, reacting slurry is taken out, after the enzyme that goes out 10min alive, carry out vacuum filtration in boiling water bath.Reducing sugar composition in filtrate is measured by the DNS method, and the enzymolysis yield is 89.81%.
Embodiment 3
Take 40 mesh sieves and dried rear corn cob 10g, add 100ml, 0.25mol/L NaOH solution be placed in beaker, and carry out ul-trasonic irradiation simultaneously, hyperacoustic power that applies is 120W, operative temperature is 25 ℃, and ultrasonic total time is 40min, after ul-trasonic irradiation 20min, adds and 3.3ml30%(v/v) H
2o
2.Reaction is carried out vacuum filtration by slurry after finishing, and realizes solid-liquid separation, and the filter residue of above-mentioned generation is carried out to the flushing of distill repeatedly water until the pH value is neutral, and dries to constant weight in 105 ℃.
The corn cob that pre-treatment is obtained carries out normal form analytical method (VanSoest) and measures (main component as shown in Figure 2) and enzymolysis, solid-to-liquid ratio 1:70(g/mL), the time is that 72h, temperature are that citric acid-sodium citrate buffer, the cellulase that 50 ℃, pH are 4.8 produced by Trichodermareesei wherein enzymatic hydrolysis condition is:, its heap(ed) capacity is the dry raw material of 20FPU/g.After enzymolysis finishes, reacting slurry is taken out, after the enzyme that goes out 10min alive, carry out vacuum filtration in boiling water bath.Reducing sugar composition in filtrate is measured by the DNS method, and the enzymolysis yield is 77.72%.
Embodiment 4
Take 40 mesh sieves and dried rear corn cob 10g, add 100ml, 0.25mol/L NaOH solution be placed in beaker, and carry out ul-trasonic irradiation simultaneously, hyperacoustic power that applies is 120W, operative temperature is 25 ℃, and ultrasonic total time is 40min, after ul-trasonic irradiation 30min, adds and 3.3ml30%(v/v) H
2o
2.Reaction is carried out vacuum filtration by slurry after finishing, and realizes solid-liquid separation, and the filter residue of above-mentioned generation is carried out to the flushing of distill repeatedly water until the pH value is neutral, and dries to constant weight in 105 ℃.
The corn cob that pre-treatment is obtained carries out normal form analytical method (VanSoest) and measures (main component as shown in Figure 2) and enzymolysis, solid-to-liquid ratio 1:70(g/mL), the time is that 72h, temperature are that citric acid-sodium citrate buffer, the cellulase that 50 ℃, pH are 4.8 produced by Trichodermareesei wherein enzymatic hydrolysis condition is:, its heap(ed) capacity is the dry raw material of 20FPU/g.After enzymolysis finishes, reacting slurry is taken out, after the enzyme that goes out 10min alive, carry out vacuum filtration in boiling water bath.Reducing sugar composition in filtrate is measured by the DNS method, and the enzymolysis yield is 78.67%.
Claims (5)
1. the method for an intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose, is characterized in that the method comprises the steps:
1) air-dry lignocellulose being carried out to the physical pulverization processing sieves again;
2) lignocellulose after screening is dried to constant weight in 105 ℃, remove wherein moisture;
3) lignocellulose after drying is added in sodium hydroxide solution and is processed, and carry out ul-trasonic irradiation simultaneously, after the 10～30min of interval, add hydrogen peroxide to continue to process, obtain pretreated reaction paste,
Wherein ultrasonic wave and coefficient total time of NaOH solution are 40～60min, and hyperacoustic operating power is 120w, and temperature is 25 ℃;
4) pretreated reaction paste in step 3) is carried out to solid-liquid separation, collect liquid material, constantly wash solid substances to pH value and be neutral and it is dried to constant weight under 105 ℃;
5) liquid material of collecting in step 4) is carried out to the recovery of hemicellulose and xylogen: utilize HCl solution that the pH value of liquid material is adjusted to 5～6, add the ethanol of 3 times of volumes, filter after standing 2h, filter residue is hemicellulose; Get its filtrate rotary evaporation that reduces pressure concentrated under 90 ℃, continue to drip HCl solution to pH value 1.5～2 after ethanol steams fully, the standing Precipitation for the treatment of, centrifugal rear gained solid is xylogen;
6) in the citric acid-sodium citrate damping fluid that is 4.5～4.8 by the solid substances after drying in step 4) in the pH value, with cellulase, be hydrolyzed;
7) slurry obtained after hydrolysis in step 6) is carried out to solid-liquid separation, liquid material is mainly the hydrolysis sugars such as glucose, and solid substances is the part hard-degraded substance.
2. the method for intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose according to claim 1, is characterized in that the described screening particle diameter of step 1) is 20 orders～60 orders.
3. the method for intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose according to claim 1, is characterized in that the described NaOH strength of solution of step 3) is 0.125～0.5mol/L, H
2o
2volume fraction be 20～40%.
4. the method for intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose according to claim 1, the solid-to-liquid ratio that it is characterized in that the described lignocellulose of step 3) and NaOH solution is 1:5～1:15g/mL, lignocellulose and H
2o
2solid-to-liquid ratio be 1:0.1～1:1g/mL.
5. the method for intensified by ultrasonic wave alkaline peroxide impregnation lignocellulose according to claim 1, it is characterized in that the described cellulase heap(ed) capacity of step 6) is the dry raw material of 5～50FPU/g, the solid-to-liquid ratio of dry raw material and citric acid-sodium citrate buffer is 1:10～1:100g/mL, hydrolysis temperature is 50 ℃, and enzymolysis time is 72h.
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