CN102399826A - Comprehensive utilizing method of sweet sorghum stalks - Google Patents
Comprehensive utilizing method of sweet sorghum stalks Download PDFInfo
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Abstract
The invention discloses a comprehensive utilizing method of sweet sorghum stalks belonging to the technical field of biomass energy and chemical industry. The method comprises the following steps of: 1) fermenting sweet sorghum stalks in a solid state and distilling the sweet sorghum stalks to obtain an ethanol solution and distillers' grains; 2) hydrolyzing the distillers' grains under an acidic condition; and carrying out solid and liquid separation to obtain an xylose solution and an acidolysis slag; 3) washing the acidolysis slag by using water as an enzymatic substrate; adding an enzymolysis buffer solution; adding enzyme to carrying out enzymolysis; and then carrying out solid and liquid separation to obtain an enzymolysis mixed solution and enzymolysis xylogen; and 4) inoculating the enzymolysis mixed solution into yeast and fermenting the enzymolysis mixed solution to generate an ethanol solution. According to the invention, a new way is found for utilizing the distillers' grains; and xylose with high value added is obtained; meanwhile, the cellulose content and cellulose accessibility in the acidolysis slag are increased so that high conversion rate of cellulose enzymolysis and saccharification is realized. By using the method disclosed by the invention, the utilization rate of the lignocellulose raw material with high sugar content is increased, the ethanol yield per unit crop is increased, and the production cost of the cellulose ethanol is reduced.
Description
Technical field
The invention belongs to biomass energy and chemical technology field, specifically, relate to the method for comprehensive utilization after a kind of sorgo stalk solid fermentation is produced ethanol.
Technical background
As energy crop, sweet sorghum has the advantage that attracts people's attention.Sweet sorghum has very high photosynthetic rate, and biological yield is high.Be rich in sugar in the stem stalk of sweet sorghum, but per hectare sweet sorghum stalk fermentative production of ethanol 3000～5000L, because of kind different.
The water requirement of planting sweet sorghum is merely 1/3～1/5 of sugarcane, and edaphic condition is had flexibility widely, particularly its significant drought resistance and saline-alkali tolerance.On marginal soils such as waste beach, punja and saltings, all can plant sweet sorghum, and 1 year three ripe.The production alcohol fuel of planting sweet sorghum has been realized " do not strive grain with the people, do not strive ground with grain ".
If plantation grain stalk dual-purpose type sweet sorghum both can be gathered in the crops seed, can gather in the crops cauline leaf again.The seed of sweet sorghum is edible both, can be used as feed and industrial raw material again; Sugar can be used the high solid-state fermentation technology (ASSF) of Tsing-Hua University's exploitation in the stem stalk, is converted into ethanol expeditiously, when producing alcohol fuel, and a large amount of vinasse of by-product.
The output of above-mentioned vinasse is much larger than alcoholic acid output.The main application of vinasse provides to livestocks such as ox, sheep as feed at present.But along with the expansion of sweet sorghum alcohol production scale, the shortage of petroleum resources more need be created bigger economic worth with the raw material of vinasse as transport fuel and Chemicals.
Summary of the invention
The objective of the invention is to provides a kind of method of comprehensive utilization of sorgo stalk to the present situation that present vinasse utilization ratio is low, the product added value is low.
The vinasse staple is semicellulose, Mierocrystalline cellulose and xylogen.Wherein, hemicellulose level is about 25%, and content of cellulose is about 35.00%.Its semicellulose component can acidolysis be used for producing important Chemicals such as wood sugar, Xylitol, furfural; After semicellulose was hydrolyzed, cellulose surface was effectively exposed in the remaining acidolysis slag, was easy to generated glucose by the cellulase degradation saccharification, and the one-step fermentation of going forward side by side is produced alcohol fuel.The remaining residue of enzymolysis is the fuel use that enzymolysis xylogen can be used as high heating value, can also deep processing production lignin product.So the comprehensive utilization of vinasse can significantly improve the economy of sweet sorghum plantation, improve the economic benefit of fuel ethanol production, alleviate the energy dilemma and the ecocrisis of China, obvious social benefit.
Based on above-mentioned principle, technical scheme of the present invention is:
A kind of sorgo stalk comprehensive utilization method comprises the steps:
(1) with solid state fermentation after sorgo stalk pulverizing, the adjusting water cut, distillation obtains ethanolic soln and vinasse;
(2) with the hydrolysis under acidic conditions of above-mentioned vinasse, solid-liquid separation obtains xylose solution and acidolysis slag;
(3) above-mentioned acidolysis pulp water is washed the back as substrate, add the enzymolysis damping fluid, add enzyme then and carry out enzymolysis, solid-liquid separation obtains enzymolysis mixed solution and enzymolysis xylogen;
(4) above-mentioned enzymolysis mixed solution inoculation yeast fermentation is produced ethanolic soln.
Realize the comprehensive utilization of sugar, Mierocrystalline cellulose, semicellulose and xylogen in the stalk composition through above-mentioned steps.Wherein, step (1) is the step of utilizing of the soluble sugar in the sorgo stalk; Step (2) is the utilization of semicellulose in the sorgo stalk and the sour preprocessing process of cellulosic ethanol; Step (3) is the process that cellulase hydrolysis produces enzymolysis mixed solution and enzymolysis xylogen in the sorgo stalk; Step (4) is the process of enzymolysis mixed solution fermentation producing and ethanol.
Pulverize described in the step (1) normally sorgo stalk is ground into diameter 1～2mm length thread less than 30mm;
The preferred method of said adjusting water cut is 70% for using deionized water to regulate the sweet Chinese sorghum pole water cut;
The condition of said solid state fermentation be 10%～20% in the yeast-inoculated amount (be every kg material used yeast liquid long-pending be 100～200mL), a temperature is 25～35 ℃ in jar, the fermentor tank velocity of rotation is under 0.2～0.5rpm condition, fermentation 20～42h.Optimum condition is: the yeast-inoculated amount is 15%, and (to be that every kg material used yeast liquid is long-pending be 150mL, contains 1 * 10 approximately in every mL yeast juice
7Individual yeast cell), temperature is 30～32 ℃ in jar, and the fermentor tank velocity of rotation is 0.25rpm, fermentation time 30h;
Said distillation is to add 150～250mL zero(ppm) water in every 50g fermentation material in the fermentation material that behind solid state fermentation, obtains, and adds thermal distillation, collects phlegma, after the phlegma of collection reaches half that adds the distillation water yield, stops.Optimum condition is: add 200mL zero(ppm) water in the every 50g fermentation material in the fermentation material that behind solid state fermentation, obtains, add thermal distillation, collect phlegma, add the distillation water yield and stop after half when the phlegma of collecting reaches.
The condition of acidolysis is in the step (2): the concentration (weight percentage) of acid is 0.5～5%, and temperature is 100～150 ℃, and the acidolysis substrate is that the weight percentage of vinasse (butt meter) is 5%～20%, acidolysis 0.1～4h.Used acid can be in sulfuric acid, phosphoric acid, formic acid, acetate, the oxalic acid one or several.
Preferred condition is: at sulfuric acid concentration (weight percentage) is 1.5%, and temperature is 120～125 ℃, and the acidolysis substrate is that the weight percentage of vinasse (butt meter) is under 10%～15% the condition, acidolysis 0.5h; Be 3% at concentration of oxalic acid (weight percentage) perhaps, temperature is 120～125 ℃, and the acidolysis substrate is that the weight percentage of vinasse (butt meter) is under 10%～15% the condition, acidolysis 1.5h.
Enzyme described in the step (3) is that cellulase or cellulase add the composite of beta-glucosidase.
Described enzymolysis is to be 4.5～5.5 in enzymolysis solution pH value; The enzymolysis substrate is that the weight percentage of acidolysis slag (butt meter) is under 5%～20% the condition, enzyme to be joined in the enzymolysis solution, and add-on is 1～10FPU/g substrate; Under 45～55 ℃ temperature, enzymatic saccharification 24～120h.
Preferred condition is: enzymolysis solution pH value is 5.0～5.1, and the enzymolysis substrate is that the weight percentage of acidolysis slag (butt meter) is 12.5%, and hydrolysis temperature is 50 ℃, enzymatic saccharification 72h.
Described solid-liquid separating method can be in separating one or more of suction filtration, centrifugal, sedimentation or spinning liquid.
The fermentation used yeast is Angel Yeast or TSH-Sc-001 bacterial classification in step (1) and (4).
TSH-Sc-001 bacterial classification according to the invention and the identical (depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center of TSH-Sc-001 bacterial classification among the patent CN101033476A; Preservation date: on March 6th, 2007, the numbering of registering on the books: 1949).
Before inoculation yeast fermentation, want in the step (4) earlier the enzymolysis mixed solution at 115～121 ℃ of 10～20min that sterilize.Fermentation condition is: the yeast-inoculated amount is that 5%～20% (volume ratio that is yeast juice and enzymolysis mixed solution is 5%～20%, contains 1 * 10 approximately in every mL yeast juice
7Individual yeast cell), temperature is 25～35 ℃ in the fermentor tank, fermentation time 20～42h.
Preferred condition is: yeast is the TSH-Sc-001 bacterial classification, and the TSH-Sc-001 inoculum size is 15%, and temperature is 28～32 ℃ in jar, fermentation time 30h.
Sweet sorghum comprehensive utilization method of the present invention, its advantage is:
1) contain 13%～14% sugar in the sorgo stalk, solid state fermentation producing and ethanol sugar utilization ratio is higher than 90%.Sweet sorghum base ethanol has been realized " do not strive grain with the people, do not strive ground with grain ".
2) solid ferment process makes that the structure of vinasse is looser, helps the carrying out of acidolysis/enzyme digestion reaction.
3) contain semicellulose about 25% in the vinasse, after diluted acid was handled, acid hydrolysis solution can be used to prepare wood sugar, and it can further generate high value added products such as Xylitol or furfural.The acidolysis process is the production process of wood sugar, also is the preprocessing process of cellulosic ethanol production simultaneously, has effectively reduced the pre-treatment cost of cellulosic ethanol production.
4) content of cellulose of vinasse acidolysis slag is high, and the cellulase accessibility is high, and the enzymolysis inhibition is few, so enzymolysis efficiency is high.Effectively reduce the enzyme cost of cellulosic ethanol production, improved the plant factor of enzymolysis process.
In a word, method of comprehensive utilization of the present invention has fully realized soluble sugar in the sorgo stalk, semicellulose and cellulosic complete utilization.Not only found new way, and obtained the wood sugar of high added value, improved content of cellulose and cellulase accessibility in the acidolysis slag simultaneously, realized the high conversion of cellulase hydrolysis saccharification for the utilization of vinasse.This method makes the utilization ratio of lignocellulosic material of high sugar degree improve, and unit crop ethanol production improves, and production costs of cellulosic ethanol descends.
Embodiment
Following embodiment can make those skilled in the art more comprehensively understand the present invention, but does not limit the present invention in any way.(annotate: the percentage ratio of following examples is weight percentage like no specified otherwise)
Embodiment 1
Gather in sophisticated sweet sorghum, with total sugar content 11%, the sweet sorghum stalk of reducing sugar content 7% is a raw material, is ground into diameter 1～2mm length thread less than 30mm, and using deionized water to regulate the sorgo stalk water cut of pulverizing is 70%.In airtight feed bin, add the TSH-Sc-001 bacterial classification, mix the back with comminuting matter and in the continuous solid-state fermentation jar, continuously ferment.The yeast-inoculated amount is that 10% (ratio that is comminuting matter and yeast juice is 1kg: 100mL, contains 1 * 10 approximately in every mL yeast juice
7Individual yeast cell), regulate each operating parameters, keeping a jar interior temperature is 30～35 ℃, and the fermentor tank velocity of rotation is 0.25rpm, fermentation time 30h.Add 200mL zero(ppm) water to every 50g fermentation material, add thermal distillation, collect phlegma, after the phlegma of collection reaches half that adds the distillation water yield, stop.
Record ethanol yield 92.2% after the fermentation, total reducing sugar transformation efficiency 98.6%.The composition of vinasse consists of: Mierocrystalline cellulose 38.43%, semicellulose 22.63%, xylogen 20.07%, ash content 0.25%.
Get 3g vinasse (dry weight) and add in the micro-wave digestion jar, add 27mL concentration again and be 3% sulfuric acid, under 130 ℃ condition, acidolysis 0.5h.Reaction finishes the back suction filtration, obtains xylose solution and acidolysis slag.
The wood sugar yield is 17.05%.The composition of acidolysis slag consists of: Mierocrystalline cellulose 45.38%, semicellulose 9.85%, xylogen 22.16%, ash content 0.68%.
Getting acidolysis slag 0.5g (dry weight) washing back joins in the triangular flask as the enzymolysis substrate; Add sodium citrate buffer, enzymolysis substrate weight percentage is that the pH value of regulating enzymolysis solution is 5.1 under 5% the condition; Cellulase is joined in the enzymolysis solution; Add-on is the 10FPU/g substrate, under 50 ℃ temperature, with the rotating speed enzymolysis of 150rpm.
Enzymolysis finishes the back solid-liquid separation and obtains enzymolysis mixed solution and enzymolysis xylogen.The cellulosic transformation efficiency of enzymatic saccharification 72h is 82.70%, and the concentration of glucose is 20.35g/L in the enzymolysis mixed solution.
At 115 ℃ of sterilization 20min, is 15% (volume ratio that be yeast juice and enzymolysis mixed solution be 15%, in every mL yeast juice approximately contain 1 * 10 in TSH-Sc-001 inoculum size then with the enzymolysis mixed solution
7Individual yeast cell), jar interior temperature is 30 ℃ condition bottom fermentation 30h.The ethanol yield reaches 90%.
In the acid hydrolysis solution in the amount of wood sugar, the enzymolysis mixed solution amount of glucose record by performance liquid chromatography.The alcoholic acid amount is recorded by gc.It all is to record by the NREL method that the one-tenth of the material among the embodiment is grouped into.
Embodiment 2
The process for solid state fermentation of sorgo stalk is with embodiment 1.
Get 3g vinasse (dry weight) and add in the microwave dissolver, adding 27mL concentration is 3% oxalic acid, under 130 ℃ condition, and acidolysis 1.5h.Reaction finishes the back suction filtration, obtains xylose solution and acidolysis slag.
The wood sugar yield is 21.48%.The composition of acidolysis slag consists of: Mierocrystalline cellulose 52.63%, semicellulose 11.67%, xylogen 20.60%, ash content 0.70%.
The enzyme solution of acidolysis slag is with embodiment 1.The cellulosic transformation efficiency of enzymatic saccharification 72h is 85.56%, and the concentration of glucose is 24.30g/L.
The fermentation condition of enzymolysis mixed solution is with embodiment 1.The ethanol yield reaches 90%.
In the acid hydrolysis solution in the amount of wood sugar, the enzymolysis mixed solution amount of glucose record by performance liquid chromatography.The alcoholic acid amount is recorded by gc.It all is to be recorded by the NREL method that the one-tenth of the material among the embodiment is grouped into.
Embodiment 3
The process for solid state fermentation of sorgo stalk is with embodiment 1.
Get 3g vinasse (dry weight) and add in the high-temperature sterilization pot, with 27mL3% oxalic acid under 125 ℃ condition, acidolysis 0.5h.Reaction finishes the back suction filtration, obtains xylose solution and acidolysis slag.
The wood sugar yield is 21.37%.The composition of acidolysis slag consists of: Mierocrystalline cellulose 51.03%, semicellulose 6.99%, xylogen 19.25%, ash content 0.73%.
The enzyme solution of acidolysis slag is with embodiment 1.The cellulosic transformation efficiency of enzymatic saccharification 48h is 65.22%, and the concentration of glucose is 18.19g/L.
The fermentation condition of enzymolysis mixed solution is with embodiment 1.The ethanol yield reaches 90%.
In the acid hydrolysis solution in the amount of wood sugar, the enzymolysis mixed solution amount of glucose record by performance liquid chromatography.The alcoholic acid amount is recorded by gc.It all is to be recorded by the NREL method that the one-tenth of the material among the embodiment is grouped into.
Embodiment 4
The process for solid state fermentation of sorgo stalk is with embodiment 1.
Get 3g vinasse (dry weight) and add in the high-temperature sterilization pot, with 27mL3% sulfuric acid under 125 ℃ condition, acidolysis 0.5h.Reaction finishes the back suction filtration, obtains xylose solution and acidolysis slag.
The wood sugar yield is 20.18%.The composition of acidolysis slag consists of: Mierocrystalline cellulose 51.95%, semicellulose 2.84%, xylogen 21.26%, ash content 0.77%.
The enzyme solution of acidolysis slag is with embodiment 1.The cellulosic transformation efficiency of enzymatic saccharification 48h is 66.05%, and the concentration of glucose is 18.47g/L.
The fermentation condition of enzymolysis mixed solution is with embodiment 1.The ethanol yield reaches 90%.
In the acid hydrolysis solution in the amount of wood sugar, the enzymolysis mixed solution amount of glucose record by performance liquid chromatography.The alcoholic acid amount is recorded by gc.It all is to be recorded by the NREL method that the one-tenth of the material among the embodiment is grouped into.
Embodiment 5
Sweet sorghum stalk is ground into diameter 1～2mm length thread less than 30mm, and using deionized water to regulate the sweet Chinese sorghum pole water cut of pulverizing is 70%.Then sorgo stalk is continuously fermented in the continuous solid-state fermentation jar, the yeast-inoculated amount is that 15% (ratio that is sorgo stalk and yeast juice is 1kg: 150mL, contains 1 * 10 approximately in every mL yeast juice
7Individual cell), regulate each operating parameters, keeping a jar interior temperature is 25～27 ℃, and the fermentor tank velocity of rotation is 0.5rpm, fermentation time 40h.Get adding 250mL zero(ppm) water in the 50g fermentation material, add thermal distillation, collect phlegma, after the phlegma of collecting reaches 125mL, stop, obtaining ethanol and vinasse.
Record ethanol yield 89.2% after the fermentation, total reducing sugar transformation efficiency 98.6%.The composition of vinasse consists of: Mierocrystalline cellulose 37.43%, semicellulose 21.63%, xylogen 22.07%, ash content 0.25%.
Get 4.5g vinasse (dry weight) and add in the micro-wave digestion jar, add 27mL concentration again and be 1.5% sulfuric acid under 120 ℃ condition, acidolysis 0.5h.Reaction finishes the back suction filtration, obtains xylose solution and acidolysis slag.
The wood sugar yield is 15.78%.The composition of acidolysis slag consists of: Mierocrystalline cellulose 45.03%, semicellulose 12.99%, xylogen 15.25%, ash content 0.73%.
Getting acidolysis slag 0.5g (dry weight) washing back joins in the triangular flask as the enzymolysis substrate; Add sodium citrate buffer, enzymolysis substrate weight percentage is that the pH value of regulating enzymolysis solution is 5.1 under 15% the condition; The prozyme of cellulase and beta-glucosidase is joined in the enzymolysis solution; Add-on is the 2FPU/g substrate, under 50 ℃ temperature, with the rotating speed enzymolysis of 150rpm.Enzymatic saccharification 48h.Reaction finishes the back suction filtration, obtains enzymolysis mixed solution and enzymolysis xylogen.The cellulosic transformation efficiency of enzymatic saccharification 48h is 49.05%, and the concentration of glucose is 25.47g/L.
At 121 ℃ of sterilization 10min, is 20% (volume ratio that be yeast juice and enzymolysis mixed solution be 20%, in every mL yeast juice approximately contain 1 * 10 in Angel Yeast inoculum size then with the enzymolysis mixed solution
7Jar individual cell), temperature is that 30 ℃ condition bottom fermentation 30h obtains ethanolic soln in.The ethanol yield reaches 90%.
In the acid hydrolysis solution in the amount of wood sugar, the enzymolysis mixed solution amount of glucose record by performance liquid chromatography.The alcoholic acid amount is recorded by gc.It all is to be recorded by the NREL method that the one-tenth of the material among the embodiment is grouped into.
Claims (10)
1. the method for comprehensive utilization of a sorgo stalk, it is characterized in that: this method comprises the steps:
(1) sorgo stalk is pulverized and regulated solid state fermentation after the water cut, distillation obtains ethanolic soln and vinasse;
(2) with the hydrolysis under acidic conditions of above-mentioned vinasse, solid-liquid separation obtains xylose solution and acidolysis slag;
(3) above-mentioned acidolysis pulp water is washed the back as substrate, add the enzymolysis damping fluid, add enzyme then and carry out enzymatic saccharification, solid-liquid separation obtains enzymolysis mixed solution and enzymolysis xylogen;
(4) with above-mentioned enzymolysis mixed solution inoculation yeast fermentative production of ethanol solution.
2. method of comprehensive utilization according to claim 1; It is characterized in that the condition of the said solid state fermentation of step (1) is: the yeast-inoculated amount is 10%～20%, temperature is 25～35 ℃ in jar; The fermentor tank velocity of rotation is 0.2～0.5rpm, and fermentation time is 20～42h; The distillatory process is: in every 50g fermentation material, add 150～250mL zero(ppm) water, add thermal distillation, collect phlegma, after the phlegma of collection reaches half that adds the distillation water yield, stop.
3. method of comprehensive utilization according to claim 1; It is characterized in that; The condition of hydrolysis under acidic conditions is described in the step (2): the concentration in acid is 0.5%～5%, and the acidolysis substrate is that the weight percentage of vinasse is 5%～20%, and temperature is under 100～150 ℃ the condition; Acidolysis 0.1～4h, wherein the weight of acidolysis substrate is in butt.
4. method of comprehensive utilization according to claim 1; It is characterized in that the condition of enzymolysis is described in the step (3): in enzymolysis solution pH value is 4.5～5.5, and the enzymolysis substrate is that the weight percentage of acidolysis slag is under 5～20% the condition; Enzyme is joined in the enzymolysis substrate; Add-on is 1～10FPU/g substrate, under 45～55 ℃ temperature, and enzymatic saccharification 24～120h.
5. method of comprehensive utilization according to claim 1 is characterized in that: the condition of fermentation is described in the step (4): the yeast-inoculated amount is 5%～20%, and temperature is 25～35 ℃ in the fermentor tank, fermentation time 20～42h.
6. method of comprehensive utilization according to claim 1 is characterized in that: wood sugar is used to produce Xylitol or furfural described in the step (2).
7. method of comprehensive utilization according to claim 1 is characterized in that: pulverizing described in the step (1) is that sorgo stalk is ground into diameter 1～2mm length thread less than 30mm.
8. method of comprehensive utilization according to claim 3 is characterized in that: used acid is one or several in sulfuric acid, phosphoric acid, formic acid, acetate, the oxalic acid in the step (2).
9. method of comprehensive utilization according to claim 1 is characterized in that: enzyme is a cellulase described in the step (3), perhaps adds the composite of beta-glucosidase for cellulase.
10. method of comprehensive utilization according to claim 1 is characterized in that: the fermentation used yeast is Angel Yeast or TSH-Sc-001 bacterial classification in step (1) and (4).
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