CN103045683B - Comprehensive utilization method of lignocellulose biomass - Google Patents
Comprehensive utilization method of lignocellulose biomass Download PDFInfo
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Abstract
A comprehensive utilization method of lignocellulose biomass comprises the following steps: (a) performing alkaline hydrolysis of the lignocellulose biomass to extract lignin; (b) performing acid hydrolysis of the residues obtained in the alkaline hydrolysis processing , separating to obtain a pentose solution and acid hydrolysis residues; (c) performing enzymatic hydrolysis of the residues obtained by alkaline hydrolysis processing in step (b) by using cellulase to obtain a solution with a main component of glucose, wherein the cellulase is obtained by culturing a strain of penicillium which has a classification name of Penicillium decumbens PD-G3-08, and is preserved in Wuhan University China Center for Type Culture Collection with a preservation number of CCTCC M 2011195 and a preservation date of June 13, 2011. The method improves the extraction ratio of cellulose enzymatic hydrolysis, and realizes comprehensive utilization of lignocellulose biomass resources.
Description
Technical field
The present invention relates to a kind of method that fully utilizes lignocellulose biomass, specifically a kind of method that fully utilizes Mierocrystalline cellulose in lignocellulose biomass, hemicellulose and xylogen.
Background technology
Day by day serious along with the increasingly exhausted and environmental pollution of fossil fuel resource, utilizes the substitute that the renewable energy resources are petroleum chemicals to become further important.And alcohol fuel is the principal mode of the material of biomass liquid energy, it is also the most probable substitute of fossil oil.At present, the mainly raw material using starch based (corn, cassava etc.) and carbohydrate (sugarcane, beet etc.) as fermentation of world's alcohol production.Adopt microbial method ferment to produce ethanol technology maturation, but high raw materials cost is restricted the industrial application of grain fermentative production of ethanol, exist simultaneously with people strive grain and grain strive etc. drawback, and cause provision price Continued, therefore find new raw material imperative.Scientist is more cheap sight trend of purchasing cost, the lignocellulose biomass widely of originating now.
Lignocellulose biomass exists with the form of plant materials, main component is Mierocrystalline cellulose, hemicellulose and xylogen, wherein, Mierocrystalline cellulose accounts for 40% left and right, hemicellulose accounts for 25% left and right, xylogen accounts for 20% left and right, and the lignocellulose biomass total amount being generated by photosynthesis every year on the earth exceedes 2,000 hundred million tons, and therefore Wooden Biomass is renewable resources abundant, the most cheap on the earth.
If can be taking lignocellulose biomass as raw material production ethanol, the mankind's energy problem will greatly be solved, but the greatest problem running in the process of raw material production ethanol taking lignocellulose biomass is that the extraction yield of cellulase hydrolysis is low, the high cost (accounting for the 40-50% of total cost of production) of enzymolysis, production cost is too high, cannot really realize industrialization.The low reason of extraction yield of cellulase hydrolysis is: hemicellulose is combined between Mierocrystalline cellulose and xylogen as molecule tamanori on the one hand, and the reticulated structure that xylogen has, surround and add set Mierocrystalline cellulose and hemicellulose as support frame, xylogen and hemicellulose spatially can hinder contacting of cellulosic molecule and enzyme, enzyme accessibility is poor, has increased the difficulty of enzymolysis.Therefore be necessary lignocellulose biomass to carry out effective pre-treatment, destroy the spatial obstacle of xylogen and hemicellulose, also to avoid pre-treatment to produce simultaneously and be unfavorable for the enzyme inhibitor (as furfural, acetic acid etc.) of enzymolysis, thereby be conducive to cellulosic enzymolysis; On the other hand, cellulase is low to crystalline cellulose enzymatic reaction vigour, therefore, in order to improve the extraction yield of cellulase hydrolysis, need to improve the vigor of cellulase.
Summary of the invention
For this reason, technical problem to be solved by this invention is to overcome prior art in the time of comprehensive utilization hemicellulose, xylogen and Mierocrystalline cellulose, the problem that cellulase extraction rate is low, thus the method for comprehensive utilization of the lignocellulose biomass that a kind of cellulase extraction rate is high is proposed.
For achieving the above object, the invention provides a kind of method of comprehensive utilization of lignocellulose biomass, the method comprises the following steps:
(a) alkaline hydrolysis: use alkaline solution treatment of lignocellulosic biomass, thereby extract xylogen, wherein said alkaline solution is aqueous sodium hydroxide solution, potassium hydroxide aqueous solution or ammoniacal liquor;
(b) acid hydrolysis: the alkaline hydrolysis residue that described alkaline hydrolysis processing is obtained carries out acid hydrolysis, obtains pentose solution and acid hydrolysis residue after separation, the acid adopting in wherein said acid hydrolysis process is one or more in sulfuric acid, hydrochloric acid, nitric acid or phosphoric acid;
(c) use cellulase to carry out enzymolysis to described acid hydrolysis residue, obtain the solution that main component is glucose; Described cellulase is to cultivate by a penicillium cellulase obtaining, this Penicillium notatum Classification And Nomenclature is Penicillium decumbens PD-G3-08, be preserved in Wuhan University's Chinese Typical Representative culture collection center, its deposit number is CCTCC M2011195, preservation date is on June 13rd, 2011, wherein, the cultural method of described cellulase is:
(1) bacterial classification multiplication culture
Be that Penicillium decumbens PD-G3-08 Penicillium notatum seed liquor is linked in the fermentor tank that contains seed culture medium through 121 DEG C of sterilizing 30min and activates with the inoculum size of percent by volume 5% by naming number, keep tank pressure 0.02-0.05MPa, air flow 0.5vvm, mixing speed 100-150rpm, 30 DEG C of cultivation 30-60 hour, obtain the seed liquor after activation;
Component in described seed culture medium and consumption are: acid hydrolysis residue 10-30g/L, wheat bran 20-50g/L in described step (b), peptone 1-4g/L, ammonium sulfate 2-4g/L, all the other are water;
(2) prepare cellulase
Step (1) is obtained to seed liquor to be accessed with the inoculum size of percent by volume 10% in the fermentor tank that 3L fermention medium is housed of sterilizing, in fermenting process, add defoamer control foaming, keep tank pressure 0.02-0.05MPa, air flow 0.5-0.6vvm, mixing speed 100-150rpm, 30 DEG C of cultivation 80-136 hour, obtain fermented liquid;
In described fermention medium, each amounts of components is respectively: acid hydrolysis residue 30-50g/L, wheat bran 20-50g/L, Microcrystalline Cellulose or carboxymethyl cellulose 4-8g/L, ammonium sulfate 2-5g/L, potassium primary phosphate 2-4g/L, magnesium sulfate 0.4-0.6g/L, all the other are water, and the initial pH of substratum is 5.0-6.0;
The fermented liquid 8000rpm centrifuging and taking obtaining obtains supernatant liquor, must contain the crude enzyme liquid of described cellulase.
Described alkaline hydrolysis carries out at 40-100 DEG C.
In described alkaline hydrolysis step, liquid-solid volume ratio is 5:1-20:1.
In described alkaline hydrolysis step, the concentration of alkaline solution is 5-8 % by weight.
The time of described alkaline solution processing is 1-6 hour.
Various alkali may be used to the present invention, includes but not limited to aqueous sodium hydroxide solution, potassium hydroxide aqueous solution, ammoniacal liquor etc.But, according to some preferred embodiment, the aqueous solution that alkaline solution is sodium hydroxide.
There is no particular limitation for the kind of described acid solution, can be that lignocellulose biomass carries out the acid-hydrolyzed conventional acid using, and for example acid can be one or more in sulfuric acid, hydrochloric acid, nitric acid and phosphoric acid.
Described acid-hydrolyzed temperature is 100-150 DEG C, time is 0.5-3 hour, while carrying out described acid hydrolysis, the concentration of acid solution is that (as the acid of selecting is strong acid to 0.5-30 % by weight, the concentration of acid solution is lower, is about 0.5-5 % by weight, as the acid of selecting is weak acid, the concentration of acid solution is higher, is about 5-30 % by weight).Described acid solution is phosphoric acid solution, and the concentration of phosphoric acid solution is 1-20 % by weight.
Described lignocellulose biomass can be one or more of maize straw, wheat straw, rice straw, bagasse, cotton bavin, cotton seed hull, corn cob, straw, kaoliang stalk, broad-leaved wood and wood chip.
Carry out pre-treatment according to raw material condition, lignocellulose biomass raw material is cut or pulverized, then this stalk section is carried out to scrubbing dust collection.
The condition of described cellulase hydrolysis is: substrate consumption is 80-150g/L, and the addition of cellulase is 10-15FPU/g Mierocrystalline cellulose, and temperature is that 45-55 DEG C, pH are that 4-6, mixing speed are 50-200rpm, and enzymolysis transformation time is 2-7 days.
After cellulase hydrolysis saccharification, fermentation can adopt the method for well known to a person skilled in the art, fermentative production of ethanol.
Technique scheme of the present invention compared with prior art has the following advantages:
1, the present invention has adopted first alkaline hydrolysis, acidolysis again, the operational path of last enzymolysis, after alkaline hydrolysis and acidolysis complete, most of hemicellulose and xylogen are extracted, Mierocrystalline cellulose comes out, thereby eliminate cellulosic spatial obstacle, made cellulase more easily contact Mierocrystalline cellulose, improved cellulosic transformation efficiency; Simultaneously because described cellulase is to cultivate by a penicillium cellulase obtaining, this Penicillium notatum Classification And Nomenclature is Penicillium decumbens PD-G3-08, be preserved in Wuhan University's Chinese Typical Representative culture collection center, its deposit number is CCTCC M2011195, the cellulase that adopts this Penicillium notatum to produce has higher vigor, has further improved the extraction yield of cellulase hydrolysis.
What 2, the present invention was used cultivates by Penicillium notatum the cellulase obtaining, be 80-150g/L at substrate consumption, the addition of cellulase is 10-15FPU/g Mierocrystalline cellulose, temperature is that 45-55 DEG C, pH are that 4-6, mixing speed are 50-200rpm, enzymolysis transformation time is that under the condition of 2-7 days, enzymolysis transformation efficiency is the highest.
3, under the temperature and time of described acid hydrolytic reaction, can be by hydrolysis of hemicellulose more thorough, can stop again under acidic conditions high temperature and reaction times long to xylogen and cellulosic destruction.
4, acid hydrolysis of the present invention acid used is phosphoric acid solution, and the concentration of phosphoric acid solution is 1-20 % by weight, uses the above-mentioned acid solution can considerable damage xylogen and Mierocrystalline cellulose, and due to phosphoric acid corrosion a little less than, therefore, maintenance of the equipment is simple, duration of service is long.
5, due to the lower extraction realizing xylogen of the alkaline solution treatment temp (40-100 DEG C) lower, reduced the destruction to xylogen.
6, the present invention adopts liquid-solid volume ratio in alkaline solution to be relatively applicable to extracting xylogen, avoid that liquid-solid ratio is too little is unfavorable for that liquid-solid mixing is also unfavorable for the alkaline hydrolysis of xylogen, it is large that the too large follow-up alkali of liquid-solid ratio reclaims load, and the wastewater flow rate of generation is also large, uneconomic problem.
7, the condition of alkaline solution processing of the present invention adopts liquid-solid ratio, alkali consumption, temperature and time, the activity that finally obtains alkali lignin is very high.
brief description of the drawings
For content of the present invention is more likely to be clearly understood, below according to a particular embodiment of the invention and by reference to the accompanying drawings, the present invention is further detailed explanation, wherein:
Fig. 1 is the schematic diagram of technical process of the present invention.
Embodiment
Below will by specific embodiment, the invention will be further described.
The self-control cellulase that following examples are used is cultivated and is obtained by Penicillium notatum, and concrete cultural method is:
(A) bacterial classification multiplication culture
Be that Penicillium decumbens PD-G3-08 Penicillium notatum seed liquor is linked in the fermentor tank that contains seed culture medium through 121 DEG C of sterilizing 30min and activates with the inoculum size of 5% (v/v) by naming number, keep tank pressure 0.02-0.05MPa, air flow 0.5vvm, mixing speed 100-150rpm, 30 DEG C of cultivation 30-60 hour, obtain the seed liquor after activation.
Component in described seed culture medium and consumption are: get embodiment 1 acid hydrolysis residue 10-30g/L, wheat bran 20-50g/L, peptone 1-4g/L, ammonium sulfate 2-4g/L, all the other are water.
Component and consumption in described seed culture medium are preferably: acid hydrolysis residue 20g/L, wheat bran 40g/L, peptone 3g/L, ammonium sulfate 3g/L, all the other are water.
(B) prepare cellulase
Step (A) is obtained to seed liquor to be accessed with the inoculum size of 10% (v/v) in the 5L fermentor tank that 3L fermention medium is housed of sterilizing, in fermenting process, add defoamer control foaming, keep tank pressure 0.02-0.05MPa, air flow 0.5-0.6vvm, mixing speed 100-150rpm, 30 DEG C of cultivation 80-136 hour, obtain fermented liquid.
In described fermention medium, each amounts of components is respectively: acid hydrolysis residue 30-50g/L, wheat bran 20-50g/L, Microcrystalline Cellulose or carboxymethyl cellulose 4-8g/L, ammonium sulfate 2-5g/L, potassium primary phosphate 2-4g/L, magnesium sulfate 0.4-0.6g/L, all the other are water, and the initial pH of substratum is 5.0-6.0.
In described fermention medium, each amounts of components is preferably: acid hydrolysis residue 45g/L, wheat bran 35g/L, Microcrystalline Cellulose 5g/L, ammonium sulfate 4g/L, potassium primary phosphate 3g/L, magnesium sulfate 0.6g/L, all the other are water, and the initial pH of substratum is 5.0-6.0.
The fermented liquid 8000rpm centrifuging and taking obtaining obtains supernatant liquor, must contain the crude enzyme liquid of cellulase, and this crude enzyme liquid can be directly used in cellulosic enzymolysis.
(2) test as follows the various performances of xylogen in following examples
The mensuration of content of lignin: comprise sour insoluble xylogen and sour solvable xylogen.Wherein the mensuration of sour insoluble xylogen adopts Klason method, carries out according to GB GB/T2677.8-94; The solvable xylogen of acid carries out according to GB GB 10337-89.
The mensuration of moisture: carry out according to GB/T 2667.3-93.
Following examples can be referring to Fig. 1.
In following examples, pressure corresponding to acid hydrolysis temperature is the pressure of saturated vapor, therefore no longer provides pressure data for each embodiment.
In following examples, outside specified otherwise, percentage composition used all represents weight percentage, i.e. " % " expression " % by weight ".
Embodiment 1
(1) alkaline solution extracts alkali lignin
Corn cob after 10.6kg is pulverized (mass component composition: moisture 6.12%, Mierocrystalline cellulose 35.19%, hemicellulose 32.1%, xylogen 23.7%, other 2.95%, lower same) smash, wash dedusting with water, then carry out alkaline hydrolysis with sodium hydroxide solution, wherein liquid-solid volume ratio is 5: 1, the concentration of sodium hydroxide solution is 6%, be warming up to 70 DEG C, through the boiling alkaline hydrolysis of 2 hours, separation obtains alkaline hydrolysis residue and alkali lignin solution, clean described alkaline hydrolysis residue with 10kg water, scavenging solution and described alkali lignin solution merge; Finally obtain the alkali lignin solution of 20.38kg alkaline hydrolysis residue (water ratio 65% left and right) and 49.61kg; In alkali lignin solution, the content of alkali lignin is 3.16%, and the extraction yield of alkali lignin is 62%.
The formula of alkali lignin extraction yield is as follows:
Alkali lignin extraction yield %=(content of xylogen in quality × alkali lignin solution of alkali lignin solution)/(content of xylogen in corn cob quality × corn cob) × 100%.
(2) acid hydrolysis
Get whole alkaline hydrolysis residues that the present embodiment (1) alkaline hydrolysis processing obtains, then be hydrolyzed with phosphoric acid solution, liquid-solid ratio is that the concentration of 10: 1 (mass ratio of new preparating acid solution and over dry alkaline hydrolysis residue) phosphoric acid solutions is 15%, carrying out acid-hydrolyzed temperature is 120 DEG C, time is 0.5 hour, acid hydrolysis residue and pentose solution that rear separation obtains are hydrolyzed, clean described acid hydrolysis residue with 10kg water, scavenging solution and described pentose solution merge, finally obtain 12.67kg acid hydrolysis residue (water ratio 65% left and right) and 75.8kg pentose solution, pentose concentration is 3.15%, hemicellulose extraction yield is 70%.
The calculation formula of hemicellulose extraction yield is as follows:
Hemicellulose extraction yield %=(concentration of the quality × pentose of pentose solution)/(content of hemicellulose in corn cob quality × corn cob) × 100%.
(3) cellulase hydrolysis
Get the described all acid hydrolytic residue of step (2) as cellulosic substrate, carry out cellulase hydrolysis, the condition of described enzymolysis is: cellulase is above-mentioned penicillin (Penicillium decumbens PD-G3-08, be preserved in Wuhan University's Chinese Typical Representative culture collection center, its deposit number is CCTCC M2011195, cultivate the cellulase obtaining down together), add cellulase according to the cellulosic addition of 15FPU/g, cellulosic substrate consumption is 150g/L, it is 48 DEG C in temperature, pH is 5.0, under the condition of mixing speed 50rpm, enzymolysis transforms 7 days, whole enzymolysis process is without pressurize, finally obtain the solution that main component is glucose, quality is 33.99kg, concentration is 7.06%, cellulosic extraction yield is 64%.
The calculation formula of Mierocrystalline cellulose extraction yield is as follows:
Mierocrystalline cellulose extraction yield %=(concentration of glucose solution quality × glucose solution)/(cellulosic content in corn cob quality × corn cob) × 100%.
The technique that glucose solution is produced ethanol is existing technique, does not repeat them here, and other embodiment is identical.
Embodiment 2
(1) alkaline solution extracts alkali lignin
Corn cob after 10.6kg is pulverized is smashed, wash dedusting with water, then carry out alkaline hydrolysis with sodium hydroxide solution, wherein liquid-solid volume ratio is 20: 1, and the concentration of sodium hydroxide solution is 5%, is warming up to 100 DEG C, through the boiling alkaline hydrolysis of 1 hour, separation obtains alkaline hydrolysis residue and alkali lignin solution, cleans described alkaline hydrolysis residue with 10kg water, and scavenging solution and described alkali lignin solution merge; Finally obtain the alkali lignin solution of 19.29kg alkaline hydrolysis residue (water ratio 65% left and right) and 200.67kg; In alkali lignin solution, the content of alkali lignin is 0.79%, and the extraction yield of alkali lignin is 63%.
(2) acid hydrolysis
Get whole alkaline hydrolysis residues that the present embodiment (1) alkaline hydrolysis processing obtains, then be hydrolyzed with phosphoric acid solution, liquid-solid ratio is that the concentration of 10: 1 (mass ratio of new preparating acid solution and over dry alkaline hydrolysis residue) phosphoric acid solutions is 10%, carrying out acid-hydrolyzed temperature is 100 DEG C, time is 1 hour, acid hydrolysis residue and pentose solution that rear separation obtains are hydrolyzed, clean described acid hydrolysis residue with 10kg water, scavenging solution and described pentose solution merge, finally obtain 11.78kg acid hydrolysis residue (water ratio 65% left and right) and 72.51kg pentose solution, pentose concentration is 3.2%, hemicellulose extraction yield is 68%.
(3) cellulase hydrolysis
Get the described all acid hydrolytic residue of step (2) as fibrous substrates, carry out cellulase hydrolysis, the condition of described enzymolysis is: cellulase is that above-mentioned penicillin (Penicillium decumbens PD-G3-08) is cultivated the cellulase obtaining, add cellulase according to the cellulosic addition of 10FPU/g, cellulosic substrate consumption is 125g/L, it is 45 DEG C in temperature, pH is 6, mixing speed is under the condition of 200rpm, enzymolysis transforms 4 days, whole enzymolysis process is without pressurize, obtain the solution that main component is glucose, quality is 32.97kg, concentration is 6.82%, cellulosic extraction yield is 60%.
Embodiment 3
(1) alkaline solution extracts alkali lignin
Corn cob after 10.6kg is pulverized is smashed, wash dedusting with water, then carry out alkaline hydrolysis with sodium hydroxide solution, wherein liquid-solid volume ratio is 10: 1, and the concentration of sodium hydroxide solution is 8%, is warming up to 40 DEG C, through the boiling alkaline hydrolysis of 6 hours, separation obtains alkaline hydrolysis residue and alkali lignin solution, cleans described alkaline hydrolysis residue with 10kg water, and scavenging solution and described alkali lignin solution merge; Finally obtain the alkali lignin solution of 21.47kg alkaline hydrolysis residue (water ratio 65% left and right) and 93.85kg; In alkali lignin solution, the content of alkali lignin is 1.53%, and the extraction yield of alkali lignin is 57%.
(2) acid hydrolysis
Get whole alkaline hydrolysis residues that the present embodiment (1) alkaline hydrolysis processing obtains, then be hydrolyzed with phosphoric acid solution, liquid-solid ratio is that the concentration of 10: 1 (mass ratio of new preparating acid solution and over dry alkaline hydrolysis residue) phosphoric acid solutions is 5%, carrying out acid-hydrolyzed temperature is 150 DEG C, time is 2 hours, acid hydrolysis residue and pentose solution that rear separation obtains are hydrolyzed, clean described acid hydrolysis residue with 10kg water, scavenging solution and described pentose solution merge, finally obtain 13.17kg acid hydrolysis residue (water ratio 65% left and right) and 79.49kg pentose solution, pentose concentration is 3.26%, hemicellulose extraction yield is 76%.
(3) cellulase hydrolysis alcohol prepared by fermenting
Get the described acid hydrolysis residue of step (2) 15.86kg as fibrous substrates, carry out cellulase hydrolysis, the condition of described enzymolysis is: cellulase is that above-mentioned penicillin (Penicillium decumbensPD-G3-08) is cultivated the cellulase obtaining, according to
12the cellulosic addition of FPU/g adds cellulase, cellulosic substrate consumption is 80g/L, be in temperature that 55 DEG C, pH are 4.0, under the condition of mixing speed 50rpm, enzymolysis transforms 2 days, whole enzymolysis process, without pressurize, obtains the solution that main component is glucose, and quality is 62.24kg, concentration is 4.15%, and cellulosic extraction yield is 69%.
Embodiment 4
(1) alkaline solution extracts alkali lignin
Wheat Straw after 11.12kg is pulverized (mass component composition: moisture 10.1%, Mierocrystalline cellulose 44%, hemicellulose 22.2%, xylogen 17%, other 6.7%) smash, wash dedusting with water, then carry out alkaline hydrolysis with sodium hydroxide solution, wherein liquid-solid volume ratio is 5: 1, the concentration of sodium hydroxide solution is 6%, be warming up to 70 DEG C, through the boiling alkaline hydrolysis of 2 hours, separation obtains alkaline hydrolysis residue and alkali lignin solution, clean described alkaline hydrolysis residue with 10kg water, scavenging solution and described alkali lignin solution merge; Finally obtain the alkali lignin solution of 21.12kg alkaline hydrolysis residue (water ratio 65% left and right) and 55.6kg; In alkali lignin solution, the content of alkali lignin is 2.04%, and the extraction yield of alkali lignin is 60%.
(2) acid hydrolysis
Get whole alkaline hydrolysis residues that the present embodiment (1) alkaline hydrolysis processing obtains, then be hydrolyzed with sulphuric acid soln, liquid-solid ratio is that the concentration of 10: 1 (mass ratio of new preparating acid solution and over dry alkaline hydrolysis residue) sulphuric acid solns is 0.5%, carrying out acid-hydrolyzed temperature is 120 DEG C, time is 3 hours, acid hydrolysis residue and pentose solution that rear separation obtains are hydrolyzed, clean described acid hydrolysis residue with 10kg water, scavenging solution and described pentose solution merge, finally obtain 15.07kg acid hydrolysis residue (water ratio 65% left and right) 76.23kg pentose solution, pentose concentration is 2.33%, hemicellulose extraction yield is 72%.
(3) cellulase hydrolysis
Get the described all acid hydrolytic residue of step (2) as fibrous substrates, carry out cellulase hydrolysis, the condition of described enzymolysis is: cellulase is that above-mentioned penicillin (Penicillium decumbens PD-G3-08) is cultivated the cellulase obtaining, add cellulase according to the cellulosic addition of 10FPU/g, cellulosic substrate consumption is 125g/L, it is 45 DEG C in temperature, pH is 6, mixing speed is under the condition of 100rpm, enzymolysis transforms 4 days, whole enzymolysis process is without pressurize, obtain the solution that main component is glucose, quality is 42.2kg, concentration is 7.3%, cellulosic extraction yield is 63%.
Found through experiments, when acid solution adopts the weak acid that concentration expressed in percentage by weight is 30%, less to xylogen and cellulosic destruction, can realize object of the present invention.And the concentration of phosphoric acid solution is while being 1%, also can realize the present invention, just needed time of acid hydrolysis and temperature of reaction need corresponding increase.
Comparative example 1
Technique and method are with embodiment 1, difference is that step (3) cellulase hydrolysis cellulase used is commercially available cellulase (jade of the He family Bioisystech Co., Ltd, 4w unit), carry out after cellulase hydrolysis, the quality of glucose solution is 33.9kg, concentration is 4.63%, and cellulosic extraction yield is 42%.
Comparative example 2
Technique and method are with embodiment 2, difference is that step (3) cellulase hydrolysis cellulase used is commercially available cellulase (jade of the He family Bioisystech Co., Ltd, 4w unit), carry out after cellulase hydrolysis, the quality of glucose solution is 32.95kg, concentration is 4.32%, and cellulosic extraction yield is 38%.
Comparative example 3
Technique and method are with embodiment 3, difference is that step (3) cellulase hydrolysis cellulase used is commercially available cellulase (jade of the He family Bioisystech Co., Ltd, 4w unit), carry out after cellulase hydrolysis, the quality of glucose solution is 62.2kg, concentration is 2.71%, and cellulosic extraction yield is 45%.
Comparative example 4
Technique and method are with embodiment 4, difference is that step (3) cellulase hydrolysis cellulase used is commercially available cellulase (jade of the He family Bioisystech Co., Ltd, 4w unit), carry out after cellulase hydrolysis, the quality of glucose solution is 42.23kg, concentration is 4.64%, and cellulosic extraction yield is 40%.
Obviously, above-described embodiment is only for example is clearly described, and the not restriction to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here without also giving exhaustive to all embodiments.And the apparent variation of being extended out thus or variation are still among the protection domain in the invention.
Claims (7)
1. a method of comprehensive utilization for lignocellulose biomass, is characterized in that the method comprises the following steps:
(a) alkaline hydrolysis: use alkaline solution treatment of lignocellulosic biomass, thereby extract xylogen, wherein said alkaline solution is aqueous sodium hydroxide solution, potassium hydroxide aqueous solution or ammoniacal liquor;
(b) acid hydrolysis: the alkaline hydrolysis residue that described alkaline hydrolysis processing is obtained carries out acid hydrolysis, obtains pentose solution and acid hydrolysis residue after separation, the acid adopting in wherein said acid hydrolysis process is one or more in sulfuric acid, hydrochloric acid, nitric acid or phosphoric acid;
(c) use cellulase to carry out enzymolysis to described acid hydrolysis residue, obtain the solution that main component is glucose; Described cellulase is to cultivate by a penicillium cellulase obtaining, and this Penicillium notatum Classification And Nomenclature is
penicillium decumbenspD-G3-08, has been preserved in Wuhan University's Chinese Typical Representative culture collection center, and its deposit number is CCTCC M 2011195, and wherein, the preparation method of described cellulase is:
(1) bacterial classification multiplication culture
By naming number be
penicillium decumbenspD-G3-08 Penicillium notatum seed liquor is linked in the fermentor tank that contains seed culture medium through 121 DEG C of sterilizing 30min and activates with the inoculum size of percent by volume 5%, keep tank pressure 0.02-0.05MPa, air flow 0.5vvm, mixing speed 100-150rpm, 30 DEG C of cultivation 30-60 hour, obtain the seed liquor after activation;
Component in described seed culture medium and consumption are: acid hydrolysis residue 10-30g/L, wheat bran 20-50g/L in described step (b), peptone 1-4g/L, ammonium sulfate 2-4g/L, all the other are water;
(2) prepare cellulase
Step (1) is obtained to seed liquor to be accessed with the inoculum size of percent by volume 10% in the fermentor tank that 3L fermention medium is housed of sterilizing, in fermenting process, add defoamer control foaming, keep tank pressure 0.02-0.05MPa, air flow 0.5-0.6vvm, mixing speed 100-150rpm, 30 DEG C of cultivation 80-136 hour, obtain fermented liquid;
In described fermention medium, each amounts of components is respectively: acid hydrolysis residue 30-50g/L, wheat bran 20-50g/L, Microcrystalline Cellulose or carboxymethyl cellulose 4-8g/L, ammonium sulfate 2-5g/L, potassium primary phosphate 2-4g/L, magnesium sulfate 0.4-0.6g/L, all the other are water, and the initial pH of substratum is 5.0-6.0;
The fermented liquid 8000rpm centrifuging and taking obtaining obtains supernatant liquor, must contain the crude enzyme liquid of described cellulase.
2. according to the method described in claim 1, it is characterized in that: described alkaline hydrolysis carries out at 40-100 DEG C.
3. method according to claim 2, is characterized in that: in described alkaline hydrolysis step, the concentration of alkaline solution is 5-8 %.
4. method according to claim 3, is characterized in that: the time of described alkaline solution processing is 1-6 hour.
5. according to the method described in claim 3 or 4, it is characterized in that: described acid-hydrolyzed temperature is 100-150 DEG C, the time is 0.5-3 hour.
6. method according to claim 5, is characterized in that: in described acid hydrolysis, the concentration of acid solution is 0.5-15 %.
7. according to the method described in claim 1,2,3,4 or 6, it is characterized in that: the condition of described enzymolysis is: substrate consumption is 80-150g/L, the addition of cellulase is 10-15FPU/g Mierocrystalline cellulose, temperature is that 45-55 DEG C, pH are that 4-6, mixing speed are 50-200rpm, and enzymolysis transformation time is 2-7 days.
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