CN105039457A - Novel process for jointly degrading and saccharifying crop straw by means of thermo-chemical treatment, microbial fermentation and enzymatic hydrolysis - Google Patents
Novel process for jointly degrading and saccharifying crop straw by means of thermo-chemical treatment, microbial fermentation and enzymatic hydrolysis Download PDFInfo
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Abstract
The invention relates to a novel process for jointly degrading and saccharifying crop straw by means of thermo-chemical treatment and enzymatic hydrolysis. The novel process includes steps of 1), mixing the smashed crop straw and soak liquid with each other according to a solid-to-liquid ratio of 1g:9-13ml to obtain mixtures, then carrying out heat-insulation treatment on the mixtures at the temperature of 70-90 DEG C for 6-18h, adding hydrogen peroxide into the mixtures after the heat-insulation treatment is completed, keeping the mixtures at the temperature of 50 DEG C for 6-18h, and fetching and drying the mixtures; 2), transferring the straw obtained in the step 1) into distilled water, adding straw with 20-60 FPU/g of cellulase and straw with 300-600U/g of xylanase into the straw, regulating the pH (potential of hydrogen) of the straw until the pH reaches 4-5, then carrying out enzymatic hydrolysis by the aid of shaking tables at the temperature of 45-50 DEG C for 24-48h, treating the straw at the temperature of 100 DEG C for 30min after enzymatic hydrolysis is completed so as to terminate enzymatic hydrolysis reaction and drying and smashing enzymatic hydrolysis products. The volume of the hydrogen peroxide is 3% of that of the soak liquid. The soak liquid is a lime supernatant with sodium hydroxide, the sodium hydroxide accounts for 2% of the weight of the crop straw, and lime accounts for 5-15% of the weight of the crop straw. The novel process has the advantage that the degradation rate of the straw and the reduced sugar yield (442.85mg/g) of the straw can be obviously increased.
Description
Technical field
The invention belongs to technical field of crop straw treatment, be specifically related to the novel process of a kind of thermochemical treatment and fermentable and enzymolysis combined degradation and saccharification agricultural crop straw.
Background technology
Straw lignocellulose is the renewable resources of the enormous amount that nature exists as structural carbon hydrate.But for want of effective treatment process, most of stalk resource is all burned as waste or is embedded in farmland.Stalk is mainly made up of Mierocrystalline cellulose, hemicellulose and xylogen, Mierocrystalline cellulose and hemicellulose can be changed into glucose monomer, ethanol, furfural, animal-feed, energy and other chemical feedstocks by fermentable and enzymolysis.But the netted aromatics polymkeric substance of the three-dimensional polymer of xylogen complexity hinders cellulase close to cellulose polymer compound.Therefore, decomposing xylogen how is gone most important to improve cellulose polymer compound enzymolysis efficiency.
At present, " people and animals strive grain " contradiction is day by day fierce, particularly in today that world population increases rapidly, how to solve this contradiction and becomes more and more important.It is feasible that Mierocrystalline cellulose in stalk and hemicellulose are transformed into the glucose energy feedstuff substituted in animal-feed as structural carbohydrate.Chang etc. (2012) and Guo etc. (2013) report that maize straw can be transformed into animal-feed by physical treatment and fermentable, and this will contribute to solving feed shortage problem in husbandry sector.
About xylogen cracking with to improve the method for stalk practicality a lot, such as acid, alkali, steam explosion and organic solvent process for stalk to improve enzymolysis efficiency.Usual high density chemical treatment (strong acid, highly basic) effect is better than low dense chemical treatment (weak acid, weak base).But stalk is developed as animal-feed, high density chemical treatment not only can damage the health of animal, also because needing a large amount of water to rinse process product, causes environmental pollution and cost increase.Forefathers report the research about bronsted lowry acids and bases bronsted lowry Treating straw production ethanol and obtain prominent achievement, but because of chemical ion (Na
+, Ca
2+and Cl
-) exceed Animal nutrition standard and be not suitable for fodder production.The organic solvents such as dimethyl sulfoxide (DMSO), DMF and 1,3-dimethyl-2-imidazolidone can be used as cosolvent and carry out Treating straw, but these process are harmful to animal health because toxic and harmful substance adds.
Summary of the invention
The object of the invention is to overcome prior art defect, provides the novel process of a kind of thermochemical treatment and fermentable and enzymolysis combined degradation and saccharification agricultural crop straw.
For achieving the above object, the present invention adopts following technical scheme:
A novel process for thermochemical treatment and enzymolysis combined degradation and saccharification agricultural crop straw, it comprises the steps:
1) after being mixed by solid-to-liquid ratio 1g:9-13ml with soak solution by the agricultural crop straw pulverized, in 70-90 DEG C of isothermal holding 6-18h, the hydrogen peroxide that isothermal holding terminates rear interpolation soak solution volume 3% keeps 6-18h in 50 DEG C, takes out, and dries; Described soak solution is the lime supernatant liquor containing sodium hydroxide, and sodium hydroxide concentration is 2% of agricultural crop straw weight, and lime is the 5-15% of agricultural crop straw weight;
2) 100ml distilled water is added in 5g stalk, step 1) gained stalk is transferred in distilled water, then cellulase 20-60FPU/g stalk and zytase 300-600U/g stalk is added, adjustment pH to 4-5, then at 45-50 DEG C of shaking table enzymolysis 24-48h, enzymolysis terminates rear 100 DEG C of process 30min with enzymolysis reaction, and enzymolysis product is dried, pulverize, to obtain final product.
A novel process for thermochemical treatment and fermentable combined degradation and saccharification agricultural crop straw, it comprises the steps:
1) after being mixed by solid-to-liquid ratio 1g:9-13ml with soak solution by the agricultural crop straw pulverized, in 70-90 DEG C of isothermal holding 6-18h, the hydrogen peroxide that isothermal holding terminates rear interpolation soak solution volume 3% keeps 6-18h in 50 DEG C, takes out, and dries; Described soak solution is the lime supernatant liquor containing sodium hydroxide, and sodium hydroxide concentration is 2% of agricultural crop straw weight, and lime consumption is the 5-15% of agricultural crop straw weight;
2) get 5g step 1) gained stalk, add 10mL, 1 × 10
6-1 × 10
7the healthy and free from worry reesei spores suspension of CFU/ml, 28-32 DEG C of fermentation culture 5-7 days., fermentation culture product is dried, and pulverizes, to obtain final product.
Preferably, the novel process of a kind of thermochemical treatment and fermentable and enzymolysis combined degradation and saccharification agricultural crop straw, it comprises the steps:
1) after being mixed by solid-to-liquid ratio 1g:9-13ml with soak solution by the agricultural crop straw pulverized, in 70-90 DEG C of isothermal holding 6-18h, the hydrogen peroxide that isothermal holding terminates rear interpolation soak solution volume 3% keeps 6-18h in 50 DEG C, takes out, and dries; Described soak solution is the lime supernatant liquor containing sodium hydroxide, and sodium hydroxide concentration is 2% of agricultural crop straw weight, and lime consumption is the 5-15% of agricultural crop straw weight;
2) get 5g step 1) gained stalk, add 10mL, 1 × 10
6-1 × 10
7the healthy and free from worry reesei spores suspension of CFU/ml, 28-32 DEG C of fermentation culture 5-7 days., fermentation culture product is dried, and pulverizes;
3) 100ml distilled water is added in 5g stalk, by step 2) gained stalk is transferred in distilled water, then cellulase 20-60FPU/g stalk and zytase 300-600U/g stalk is added, adjustment pH to 4-5, then at 45-50 DEG C of shaking table enzymolysis 24-48h, enzymolysis terminates rear 100 DEG C of process 30min with enzymolysis reaction, and enzymolysis product is dried, pulverize, to obtain final product.
Described agricultural crop straw can be maize straw, wheat stalk etc.The application studies for maize straw.
The drawback existed in existing treatment process, the application is according to NRC(1994) Animal nutrition standard-required, select low-concentration sodium hydroxide and lime to process maize straw.Because Ca
2+and Na
+the required mineral element of animal, in Animal nutrition standard about poultry to Ca
2+and Na
+demand is respectively 0.8-2.0% and 0.12-0.2%.Ca residual in chemical treatment stalk
2+and Na
+can be absorbed by animal, to reduce the interpolation of stone flour and salt in diet recipe.Pyroprocessing (> 100 DEG C) is strict to equipment requirements, portion of cellulose and hemicellulose resolve into reducing sugar, the by products such as furfural, formic acid and propionic acid can be changed into further, not only affect animal health, also produce restraining effect to enzymolysis, therefore the application selects treatment temp <100 DEG C.The application obtains optimum lime concentration and treatment condition by response surface design, and carries out single and compound comparative analysis to lime, sodium hydroxide and hydrogen peroxide.In addition, for further improving degradation rate and the conversion coefficient of stalk, the condition of fermentable and enzymolysis is also studied.Result shows, maize straw is immersed in the mixing solutions of 15% (w/w) lime supernatant liquor and 2% (w/w) sodium hydroxide composition with liquid-solid ratio 13:1 (mL/g), under 83.92 DEG C of conditions, keep 6h; Add 3% (v/v) H subsequently
2o
2, under 50 DEG C of conditions, keep 2h; Finally add cellulase (32.3FPU/g dry-matter) and zytase (550U/g dry-matter), 50 DEG C of shaking table enzymolysis 48h, obtaining maximum reducing sugar output is 348.77mg/g.Compared with original enzymolysis stalk, after chemical treatment enzymolysis again stalk in reducing sugar output increased 126.42% (P<0.05), Mierocrystalline cellulose, hemicellulose and Lignin degradation rate improve 40.08% (P<0.05) respectively, 45.71% (P<0.05) and 52.01% (P<0.05).Stalk after chemical treatment, then through fermentable and enzymolysis combination treatment, can further improve degradation rate and the reducing sugar output (442.85mg/g, P<0.05) of stalk.
Accompanying drawing explanation
Fig. 1 is the scanning electron microscope image of different maize straw sample; In figure, (A) untreated maize straw; (B) chemical treatment (15%CaO+2%NaOH+3%H
2o
2) maize straw; (C) the mould fermentation of chemical treatment+healthy and free from worry wood+enzymolysis combination treatment stalk.
Embodiment
Do to introduce in detail further to technical scheme of the present invention below in conjunction with embodiment, but protection scope of the present invention is not limited thereto.
Embodiment
1. materials and methods
1.1 material
Natural air drying maize straw is pulverized through beater disintegrating machine, and sieve 1mm, room temperature preservation.Cellulase and zytase are purchased from Shandong Ze Sheng Bioisystech Co., Ltd.323FPU/g and 5500U/g is respectively according to U.S.'s renewable energy source laboratory (NREL) cellulase measuring method mensuration cellulase activity and xylanase activity.Healthy and free from worry wood is mould purchased from China General Microbiological culture presevation administrative center, bacterium numbering CGMCC3.4262.
1.2 optimum chemical treatment condition response surface designs
Utilize Design-Expert8.0.5 software, adopt Box-Behnken to be designed for test design, model-fitting and data analysis.Test is got in 15g maize straw loading 500ml beaker and is sealed with masking foil, carries out isothermal holding with water-bath.Response surface optimization is based on single factor test result in early stage, and each selecting factors scope is treatment temp 70-90 DEG C; Treatment time 6-18h; The concentration 5-15%(w/w of lime supernatant liquor, lime/stalk); Liquid-solid ratio is 9:1-13:1, and experimental factor and water-glass are in table 1.While lime treatment, add 2% sodium hydroxide (w/w, sodium hydroxide/stalk), the selection of naoh concentration is to Na according to poultry in Animal nutrition standard
+demand.After lime and sodium-hydroxide treatment terminate, add 3% hydrogen peroxide (v/v), 50 DEG C keep 2h, take out 65 DEG C of oven dry, for enzymolysis analysis.
Table 1 test design factor and code levels table
Being prepared as follows of different concns lime supernatant liquor: the often heavy 15.0g of the air-dry straw sample of group process, according to the percent by weight of stalk, get 5-15% lime (w/w, lime/stalk), be dissolved according to different liquid-solid ratio 9:1-13:1(water: stalk) in the water got, stir 5min, static 10min, gets supernatant liquor.
In order to embody the relation of independent variable(s) and dependent variable, adopt the multinomial equation of secondary to carry out matching, the multinomial equation form of prediction secondary is as follows:
Y=β
0
+β
1
X
1
+β
2
X
2
+β
3
X
3
+β
4
X
4
+β
11
X
1
2
+β
22
X
2
2
+β
33
X
3
2
+β
44
X
4
2
+β
12
X
1
X
2
+β
13
X
1
X
3
+β
14
X
1
X
4
+β
23
X
2
X
3
+β
24
X
2
X
4
+β
34
X
3
X
4；
In formula:
ypredict reducing sugar content (mg/g dry-matter) after chemical treatment stalk enzymolysis;
x 1 ,
x 2 ,
x 3 ,
x 4 independent variable(s), temperature of reaction corresponding respectively, time, lime concentration, liquid-solid ratio;
β 0 it is intercept;
β 1,
β 2,
β 3,
β 4 it is linear coefficient;
β 11,
β 22,
β 33,
β 44 it is squared modulus;
β 12,
β 13,
β 14 ,
β 23,
β 24 ,
β 34 it is interaction coefficent.
1.3 single and composite chemical Treating straws
This test carries out on above-mentioned response surface design gained optimal treatment condition basis, and treatment condition are liquid-solid ratio 13:1, and 84 DEG C keep 6h.Test design is grouped as follows:
1 group: untreated stalk;
Lime supernatant liquor Treating straw 2 groups: 15%(w/w);
Sodium-hydroxide treatment stalk 3 groups: 2%(w/w);
Lime supernatant liquor+2%(w/w 4 groups: 15%(w/w)) sodium-hydroxide treatment stalk;
Lime supernatant liquor+3% (v/v) hydrogen peroxide Treating straw 5 groups: 15%(w/w);
Sodium hydroxide+3% (v/v) hydrogen peroxide Treating straw 6 groups: 2%(w/w);
Lime supernatant liquor+2%(w/w 7 groups: 15%(w/w)) sodium hydroxide+3% (v/v) hydrogen peroxide Treating straw;
Hydrogen peroxide Treating straw 8 groups: 3%(v/v).
1.4 cellulases and zytase enzymolysis processing
Enzymolysis processing is with 5%(w/v) stalk is added in distilled water after chemical treatment, adds cellulase 32.3FPU/g stalk and zytase 550U/g stalk, with 0.05MHCL or Ca (OH)
2supernatant liquor is adjusted to pH4.8, shaking table (180rpm) enzymolysis 48h under 50 DEG C of conditions, and enzymolysis terminates rear 100 DEG C of process 30min with enzymolysis reaction, and enzymolysis product 50 DEG C oven dry, pulverizes.
The mould fermentation of 1.5 healthy and free from worry wood and the mould fermentation of healthy and free from worry wood and enzymolysis combination treatment
After getting 5.0g chemical treatment, stalk loads in the triangular flask of 250mL band tampon, adds the healthy and free from worry wood of 10mL mould (CGMCC3.4262) spore suspension (1 × 10
6cFU/ml), cultivate 5d., fermentation ends 50 DEG C oven dry for 30 DEG C, pulverize.
Tunning carries out enzymolysis again, and enzymolysis processing is with above-mentioned 1.4.Test is grouped as follows:
I group: chemical treatment (NaOH+Ca (OH)
2+ H
2o
2) stalk
II group: chemical treatment+mould fermented stalk of healthy and free from worry wood
III group: chemical treatment+enzymolysis stalk
IV group: the mould fermentation of chemical treatment+healthy and free from worry wood+enzymolysis combination treatment stalk
The mensuration of reducing sugar and moiety in 1.6 stalks
Mierocrystalline cellulose, hemicellulose and xylogen measure and adopt VanSoest method; Total reducing sugars measures employing 3,5-dinitrosalicylic acid (DNS) method.Index reduction formula is as follows:
Y=
X×
W 1 /
W 2 ,
In formula:
yrepresent the content relative to each index in stalk before process;
xrepresent the content of each index in Treating straw;
w 1 represent the rear stalk dry matter weight of process;
w 2 represent the front stalk dry matter weight of process.
The preparation of 1.7 stalk electron microscopic samples
Choose untreated stalk, chemical treatment stalk and the mould fermentation of chemical treatment+healthy and free from worry wood+enzymolysis combination treatment straw sample 2.5% glutaraldehyde solution respectively and fix more than 4h, 0.1mol/L phosphoric acid buffer rinses 3 times, 30%, 50%, 70%, 80%, 90%, 100% ethanol dewaters step by step, every grade of dehydration 15min, then 2 times are replaced with isoamyl acetate, each 10min.Take out rearmounted dry basket and put into rapidly EMITECHK850 critical point drying instrument, take out after dry, stick in sample table with conductive resin, vacuum metal spraying plated film is carried out with HITACHIE 1 type ion sputtering instrument, under being placed in HITACHIS-3400N II type scanning electron microscope, observe under 1000 × and 2000 × enlargement ratio.
1.8 data statisticss and analysis
Testing data all represents with " mean+SD " (mean ± SD), and in result SPSS20.0 statistical software, ANOVA process carries out variance analysis, and carries out Duncan multiple comparisons,
p<0.05 is significant difference.
2 results and discussion
2.1 optimal treatment condition response surface analysises
According to Box – Behnken test design in DesignExpert8.0.5 software, devise the response surface analysis test of 27 test points, after obtaining pre-treatment to test, reducing sugar content returns, set up response surface quadratic regression model, seek the level of factor of optimal response value, test-results and regression equation variance analysis are in table 2 and table 3.
Table 2Box-Behnken design and result
The test of significance of table 3 regression equation coefficient
Design-Expert8.0.6 carries out polynary Quadratic Regression Fitting to data, draws regression model equation and the results of analysis of variance, as follows:
Y＝－683.09＋9.03
X 1 ＋24.33
X 2 ＋11.20
X 3 ＋62.04
X 4 -0.049
X 1 2 ＋0.050
X 1 X 2 ＋0.13
X 1 X 3 －0.24
X 1 X 4 －0.24
X 2 2 －0.92
X 2 X 3 －1.30
X 2 X 4 －0.27
X 3 2 －0.23
X 3 X 4 －0.82
X 4 2 。
As can be seen from Table 3, above-mentioned model
p<0.0001, show that this modeling statistics is upper meaningful.Conspicuous level (
p<0.05) under condition, temperature in total reducing sugars regression model
x 1 (
p=0.0147), the time
x 2 (
p=0.0227), lime concentration
x 3 (
p<0.0001), liquid-solid ratio
x 4 (
p<0.0001), show that these factors have remarkably influenced to response value;
x 2 x 3 (
p<0.0001),
x 2 x 4 (
p=0.0003) performance significantly, shows in reaction times and lime concentration, there is interaction between reaction times and liquid-solid ratio.Lose and intend item
pvalue is intended not remarkable for 0.147>0.005 and equation model lose, illustrate that the degree of fitting of equation is better.The coefficient of determination of regression model is R
2=0.9554>0.80 and R
2 adj=0.9035>0.80, the coefficient of determination, more close to 1, illustrates that the degree of fitting of regression equation is better, can the change of interpretation model preferably.Therefore, this model available carries out analysis and prediction to maize straw pretreating effect.
Response surface figure and corresponding isogram, each 3D response surface chart is shown in wherein that two factors are under the condition of 0 level, and two other factor is to the interact relation of response value reducing sugar.Based on response surface analysis, by software simulation optimizing, when temperature 83.92 DEG C, reaction times 6h, 15%(w/w) lime supernatant liquor, during liquid-solid ratio 13:1, total reducing sugars content reaches the highest, predict the outcome as 342.34mg/g, basically identical with trial value 348.77mg/g, show that Optimized model has higher reliability.
2.2 different chemical process are on the impact of the main moiety of maize straw
In different chemical process and enzymolysis stalk, the content of Mierocrystalline cellulose, hemicellulose and xylogen is in table 4.Compared with untreated stalk, except 2 and 3 groups, in other each group after chemical treatment in stalk content of cellulose significantly improve (
p<0.05), and hemicellulose and content of lignin significantly reduce (
p<0.05), this shows that lower concentration lime or sodium hydroxide individual curing stalk are removed without remarkable effect xylogen, its Main Function is swollen xylogen and Mierocrystalline cellulose internal structure, the 3%(v/v followed closely) hydrogen peroxide process has very strong delignification.Lower concentration lime and sodium hydroxide combination treatment are significantly higher than individual curing to xylogen removal effect, and this shows that lime and sodium hydroxide combination treatment have positive-effect.
Compared with untreated enzymolysis stalk, in 4th group of chemical treatment enzymolysis stalk, Mierocrystalline cellulose, hemicellulose and Lignin degradation rate improve 31.86% (P<0.05) respectively, 41.04% (P<0.05) and 2.49% (P>0.05); In 7th group of chemical treatment enzymolysis stalk, Mierocrystalline cellulose, hemicellulose and Lignin degradation rate improve 40.08% (P<0.05) respectively, 45.71% (P<0.05) and 52.01% (P<0.05).This shows that lime, sodium hydroxide and hydrogen peroxide combination treatment can improve lignin removing rate and enzymatic saccharification rate.
The main moiety of table 4 different chemical process maize straw (%, dry-matter)
Remarks: 1 group: untreated maize straw; 2 groups: 15% lime supernatant liquid; 3 groups: 2% sodium hydroxide; 4 groups: 15% lime supernatant liquid+2% sodium hydroxide; 5 groups: 15% lime supernatant liquid+3% hydrogen peroxide; 6 groups: 2% sodium hydroxide+3% hydrogen peroxide; 7 groups: 15% lime supernatant liquid+2% sodium hydroxide+3% hydrogen peroxide; 8 groups: 3% hydrogen peroxide.Y1 ' represents wood fibre cellulose content after chemical treatment; Y2 ' represents that chemical treatment adds wood fibre cellulose content after enzymolysis.The identical person's difference of same column mark capitalization is not significantly (P>0.05); Capitalization difference person significant difference (P<0.05).
2.3 different chemical process are on the impact of stalk pH and reducing sugar content
Reduce content in table 5 before and after the change of pH and enzymolysis before and after chemical treatment.Test-results shows, and except the 1st group, after chemical treatment, stalk pH is lower than before process.After chemical treatment reducing sugar content significantly lower than untreated stalk (
p<0.05).Its reason is that complicated reaction can occur reducing sugar in hot alkaline solution, as isomerization, decomposition and redox generation carboxylic acid under the condition of oxygenant, this shows that alkali and hydrogen peroxide process can be not only that hemicellulose and xylogen decompose, and original reducing sugar content in stalk also can be made to reduce.
Reducing sugar in chemical treatment enzymolysis stalk is decomposed by Mierocrystalline cellulose and hemicellulose to obtain, and chemical treatment mainly improves the contact area of Mierocrystalline cellulose and hemicellulose and enzyme.Compared with untreated enzymolysis stalk, in 2-7 group in chemical treatment enzymolysis stalk reducing sugar content be significantly increased (
p<0.05), improve 59.90%, 20.18%, 94.12%, 73.67%, 42.13% and 126.42% respectively, and in lime, sodium hydroxide and hydrogen peroxide combination treatment enzymolysis stalk (the 7th group), reducing sugar content is the highest.This shows that alkali and hydrogen peroxide combination treatment effect are better than individual curing.Its reason is that alkali swollen cellulosic molecule internal structure makes cellulosic molecule become porous, reduces cellulose crystallity, the contact area of increase and enzyme.Another reason is that lime has the effect of removing enzymolysis supressor mixture, increases the activity of enzyme.
The change (mg/g dry-matter) of table 5 maize straw pH change and reducing sugar content after different chemical process and after enzymolysis
Remarks: 1 group: untreated maize straw; 2 groups: 15% lime supernatant liquid; 3 groups: 2% sodium hydroxide; 4 groups: 15% lime supernatant liquid+2% sodium hydroxide; 5 groups: 15% lime supernatant liquid+3% hydrogen peroxide; 6 groups: 2% sodium hydroxide+3% hydrogen peroxide; 7 groups: 15% lime supernatant liquid+2% sodium hydroxide :+3% hydrogen peroxide; 8 groups: 3% hydrogen peroxide.Y1 ' represents wood fibre cellulose content after chemical treatment; Y2 ' represents that chemical treatment adds wood fibre cellulose content after enzymolysis.The identical person's difference of same column mark capitalization is not significantly (P>0.05); Capitalization difference person significant difference (P<0.05).
2.4 fermentable and enzymolysis are on the impact of reducing sugar output
Chemical treatment stalk after fermentable, enzymolysis and the two combination treatment Mierocrystalline cellulose, hemicellulose, xylogen and reducing sugar content in table 6.Result shows, chemical treatment stalk Mierocrystalline cellulose, hemicellulose, content of lignin after fermentable, enzymolysis and the two combination treatment significantly reduce (
p<0.05).Chemical treatment stalk reducing sugar content after fermentable and enzyme combination treatment be significantly higher than fermentable and enzymolysis individual curing (
p<0.05).This show stalk through chemical treatment destroy xylogen densification structure and to cellulosic provide protection, reduce the granular size of lignocellulose, in increase, the contact area of Mierocrystalline cellulose and microorganism and enzyme, is easy to enzymolysis.
The change (%) of table 6 chemical treatment stalk reducing sugar and wood fibre cellulose content after different enzymolysis processing
Remarks: I group: chemical treatment stalk; II group: chemical treatment+mould fermented stalk of healthy and free from worry wood; III group: chemical treatment+enzymolysis stalk; IV group: the mould fermentation of chemical treatment+healthy and free from worry wood+enzymolysis combination treatment stalk.The identical person's difference of same column mark capitalization is not significantly (P>0.05); Capitalization difference person significant difference (P<0.05).
2.5 maize straw surface morphological structure are observed
The morphological structure of untreated stalk, chemical treatment stalk and chemical treatment+fermentable+enzymolysis combination treatment stalk is shown in 1.Untreated stalk color is tawny, amplifies 1000 times of observations (in Fig. 1 A) through scanning electron microscope, and xylogen smooth surface is fine and close, exposes without hemicellulose and Mierocrystalline cellulose.After 15% lime+2% sodium hydroxide+3% hydrogen peroxide process, maize straw color is light yellow, amplify 2000 times of observations (in Fig. 1 B) through scanning electron microscope, xylogen is damaged, peel off, Mierocrystalline cellulose vascular bundle obviously exposes, and formed in many " hole ", specific surface area increases.It mainly contains the oxidation delignification that reason is hydrogen peroxide, especially in the basic conditions.Chemical treatment stalk is frangible after fermentable and enzymolysis combination treatment, and color becomes brown, and amplify 2000 times of observations (in Fig. 1 C) through scanning electron microscope, Mierocrystalline cellulose vascular bundle is broken down into fragment.Mierocrystalline cellulose vascular bundle is easily decomposed mainly because the destruction of xylogen complex construction and cellulosic porousness by microorganism and enzyme, makes enzyme be easy to attack Mierocrystalline cellulose, improves reducing sugar content, and proves reasoning above.
3. conclusion
Utilize Responds Surface Methodology to establish with reducing sugar to be the technological mathematical model of response value, and carried out process optimization simulation on this basis, draw the optimal treatment condition of lime+2% sodium hydroxide: temperature 83.92 DEG C, time 6h, lime concentration 15%, liquid-solid ratio 13:1, actually obtains enzymolysis reducing sugar 348.77mg/g.By single and composite chemical Treatment Analysis, lime, sodium hydroxide and hydrogen peroxide combination treatment, and carry out fermentable and enzyme associating saccharification process, to the removal of stalk cellulose, hemicellulose and xylogen with to improve conversion coefficient effect comparatively remarkable, for practical application provides theoretical foundation.
Claims (3)
1. a novel process for thermochemical treatment and enzymolysis combined degradation and saccharification agricultural crop straw, is characterized in that, comprises the steps:
1) after being mixed by solid-to-liquid ratio 1g:9-13ml with soak solution by the agricultural crop straw pulverized, in 70-90 DEG C of isothermal holding 6-18h, the hydrogen peroxide that isothermal holding terminates rear interpolation soak solution volume 3% keeps 6-18h in 50 DEG C, takes out, and dries; Described soak solution is the lime supernatant liquor containing sodium hydroxide, and sodium hydroxide concentration is 2% of agricultural crop straw weight, and lime consumption is the 5-15% of agricultural crop straw weight;
2) 100ml distilled water is added in 5g stalk, step 1) gained stalk is transferred in distilled water, then cellulase 20-60FPU/g stalk and zytase 300-600U/g stalk is added, adjustment pH to 4-5, then at 45-50 DEG C of shaking table enzymolysis 24-48h, enzymolysis terminates rear 100 DEG C of process 30min with enzymolysis reaction, and enzymolysis product is dried, pulverize, to obtain final product.
2. a novel process for thermochemical treatment and fermentable combined degradation and saccharification agricultural crop straw, is characterized in that, comprises the steps:
1) after being mixed by solid-to-liquid ratio 1g:9-13ml with soak solution by the agricultural crop straw pulverized, in 70-90 DEG C of isothermal holding 6-18h, the hydrogen peroxide that isothermal holding terminates rear interpolation soak solution volume 3% keeps 6-18h in 50 DEG C, takes out, and dries; Described soak solution is the lime supernatant liquor containing sodium hydroxide, and sodium hydroxide concentration is 2% of agricultural crop straw weight, and lime consumption is the 5-15% of agricultural crop straw weight;
2) get 5g step 1) gained stalk, add 10mL, 1 × 10
6-1 × 10
7the healthy and free from worry reesei spores suspension of CFU/ml, 28-32 DEG C of fermentation culture 5-7 days, fermentation culture product is dried, and pulverizes, to obtain final product.
3. a novel process for thermochemical treatment and fermentable and enzymolysis combined degradation and saccharification agricultural crop straw, is characterized in that, comprises the steps:
1) after being mixed by solid-to-liquid ratio 1g:9-13ml with soak solution by the agricultural crop straw pulverized, in 70-90 DEG C of isothermal holding 6-18h, the hydrogen peroxide that isothermal holding terminates rear interpolation soak solution volume 3% keeps 6-18h in 50 DEG C, takes out, and dries; Described soak solution is the lime supernatant liquor containing sodium hydroxide, and sodium hydroxide concentration is 2% of agricultural crop straw weight, and lime consumption is the 5-15% of agricultural crop straw weight;
2) get 5g step 1) gained stalk, add 10mL, 1 × 10
6-1 × 10
7the healthy and free from worry reesei spores suspension of CFU/ml, 28-32 DEG C of fermentation culture 5-7 days, fermentation culture product is dried, and pulverizes;
3) 100ml distilled water is added in 5g stalk, by step 2) gained stalk is transferred in distilled water, then cellulase 20-60FPU/g stalk and zytase 300-600U/g stalk is added, adjustment pH to 4-5, then at 45-50 DEG C of shaking table enzymolysis 24-48h, enzymolysis terminates rear 100 DEG C of process 30min with enzymolysis reaction, and enzymolysis product is dried, pulverize, to obtain final product.
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| CN105385724A (en) * | 2015-12-24 | 2016-03-09 | 齐鲁工业大学 | Method for improving conversion efficiency of lignocellulose through combined treatment and method for efficiently preparing ethyl alcohol |
| CN106011199A (en) * | 2016-07-19 | 2016-10-12 | 南京林业大学 | Method for pre-treating crop straw |
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| CN106011199B (en) * | 2016-07-19 | 2020-01-10 | 南京林业大学 | Pretreatment method of crop straws |
| CN106011199A (en) * | 2016-07-19 | 2016-10-12 | 南京林业大学 | Method for pre-treating crop straw |
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| CN107012195B (en) * | 2017-06-12 | 2022-07-12 | 北京中农富源生物工程技术有限公司 | Method for producing hydrogen by fermenting crop straws as raw materials |
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