CN103060418A - Method of constructing mixed bacteria system for fermenting straw stalks to produce ethanol - Google Patents

Method of constructing mixed bacteria system for fermenting straw stalks to produce ethanol Download PDF

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CN103060418A
CN103060418A CN2012105170029A CN201210517002A CN103060418A CN 103060418 A CN103060418 A CN 103060418A CN 2012105170029 A CN2012105170029 A CN 2012105170029A CN 201210517002 A CN201210517002 A CN 201210517002A CN 103060418 A CN103060418 A CN 103060418A
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straw
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rice straw
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徐尔尼
巫小丹
徐颖宣
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Nanchang University
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Abstract

The method provides a method of totally utilizing a microbial technology to realize conversion of straw stalks into ethanol. The method uses mixed bacteria of Trichoderma viride and Trichoderma reesei which can produce high-efficient cellulase and complementary lignin degradation enzymes to ferment the straw stalks, thereby realizing synchronous production of the cellulase and the lignin degradation enzymes, achieving effects of pretreatment for the straw stalks by microorganism, and through a variable-temperature technology, realizing in situ enzymatic saccharification for the straw stalks by the lignin degradation enzymes produced by the microorganism; finally, the method uses an SA-PVA-SiO2 immobilization technology to mix and embed in a co-immobilization way saccharomyces cerevisiae capable of fermenting hexose to produce the ethanol and candida tropicalis capable of fermenting pentose to produce the ethanol, and constructs a microenvironment suitable for fermentation of the microzyme so as to produce the ethanol, thereby improving an efficiency of ethanol production, being low in energy consumption, non-polluted, and short in production period, and realizing recycling of the microzyme.

Description

A kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw
Technical field
The present invention relates to a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw.
Background technology
Now, the socio-economic development of countries in the world is higher to the interdependency of petroleum resources, and that oil has is non-renewable, will have influence on for a long time the Sustainable development of society.China is oil consumption big country, but the domestic petroleum resource is relatively poor.The development and use new and renewable sources of energy has become world's common recognition, and becomes the national strategy of China's adjustment, Optimization of Energy Structure.Ethanol petrol because of have sufficient combustion, pollution products few, can play certain advantages such as explosion-proof and praised highly, alcohol fuel industry faces new opportunity.The national conditions of having a large population and a few land based on China, food crop obviously no longer are suitable for the raw material sources as the exploitation alcohol fuel, but as large agricultural country, China's agricultural crop straw (biomass) annual production occupies first of the world, be about 7.4 hundred million ton/years, the stalk that wherein can be used as the fuel ethanol production raw material accounts for 20% of agricultural crop straw total amount, reaches 1.4 hundred million tons, can provide stable raw material sources for China produces alcohol fuel.Utilize biomass to be the raw material production alcohol fuel; be the reality needs of alleviating energy crisis; can effectively solve current energy scarcity problem; it also is the inevitable choice of preserving the ecological environment; will greatly reduce waste to the pollution of environment; can also alleviate the crisis in food that traditional method utilizes the starch ethanol conversion to cause, the Sustainable development of China's economic society is of great immediate significance and far-reaching historic significance.
The main component of rice straw is xylogen (content is about 20%~25%), hemicellulose (content is about 25%~30%) and Mierocrystalline cellulose (content is about 30%~35%).Need to remove the steric restriction effect of xylogen before the rice straw enzymatic saccharification, in the To Several Traditional Preconditioning Methods, large, the weak effect of physical method power consumption, chemical process easily causes secondary pollution.The microbial enzyme facture has the advantages such as pollution-free, that cost is low, and the microbiological deterioration xylogen depends on the acting in conjunction of lignin peroxidase and laccase.Research finds that the strong microorganism of product lignin-degrading enzymes ability is less, and the lignocellulolytic enzymes of different strains is variant, and mixed fungus fermentation can remedy the difference between the bacterial classification, gives full play to the synergy between each enzyme.Studies show that a kind of viride ( Trichoderma viride) laccase production ability is very strong, the laccase that produces can make the methyl catechol generation polymerization in the PDA substratum that has added 0.04% methyl catechol, produces the sorrel compound, form the sorrel zone of oxidation, so the size of zone of oxidation can be indicated the size of laccase production ability.Viride is cultivated 2 d can form the sorrel zone of oxidation that diameter reaches 3.0~4.0 cm in methyl catechol-PDA substratum.By contrast, be usually used in the pretreated flat mushroom of stalk and cultivate the sorrel zone of oxidation that 7 d ability formation diameter in methyl catechol-PDA substratum reaches 3.0 cm.Research discovery Trichodermareesei ( Trichoderma reesei) to produce the lignin peroxidase ability very strong, the lignin peroxidase that produces can make the PDA substratum generation color fading reaction that has added 0.01% aniline blue, the formation transparent circle, so the size of transparent circle can be indicated the size of product lignin peroxidase ability.Trichodermareesei is cultivated 2 d can form the transparent circle that diameter reaches 4.0 cm at aniline blue-PDA substratum, and 3 d can make whole flat board fade fully.By contrast, being usually used in the pretreated flat mushroom of stalk cultivates 4 d and just forms the transparent circle that diameter reaches 2.0~3.0 cm at aniline blue-PDA substratum.Trichodermareesei and viride have complementary Ligninolytic Enzymes, and the two mixed culture can reach preferably rice straw pretreating effect, and the cellulase of the two, hemicellulase are lived also stronger.
85%~90% of natural hemicellulose hydrolysate, wood sugar.Produce ethanol with the wood-sugar fermentation in the plant cellulose material, can realize taking full advantage of of raw material, improve the efficient of stalk ethanol conversion, reduce production costs.Traditional alcohol production bacterial strain yeast saccharomyces cerevisiae ( Saecharomyces cerevisiae) unfermentable wood sugar, but some yeast can xylose fermentation ethanol, as candida tropicalis ( Candida tropicalis), pipe capsule yeast ( Pachysolen tannophilus), pichia stipitis ( Pichia stipitis), shehatae candida ( Candida shehatae) and heat-resisting methylotrophic yeast ( Hansenula polymorpha) etc.Mainly contain glucose and xylose in the rice straw enzymatic saccharification liquid, the ethanol conversion that yeast saccharomyces cerevisiae and yeast for fermentation straw saccharification liquid that can xylose fermentation ethanol can the Effective Raise rice straws.
Ethanol fermentation intensity directly affects the efficient of stalk ethanol conversion, and ethanol fermentation intensity depends mainly on saccharomycetic kind and cell concn.Originally traditional sequestered yeast exists that yeast cell concentration is on the low side, cell proliferation period is long and the problem such as product collection stage yeast loss in fermentation producing and ethanol process.The immobilized cell technology that grow up the sixties in 20th century have can be continuously with reuse, can realize the characteristics such as cell high-density culture, product separation are simple.Process for fixation comprises entrapping method, Chemical bond method, physisorphtion, wherein entrapping method is to use very wide a kind of process for fixation, its formed immobilization microenvironment is fit to saccharomycetic anaerobically fermenting very much, and has the advantages such as alcohol getting rate height, strong, the anti-high glucose concentration of acetic acid tolerance and contamination resistance are strong.Alginate calcium is as a kind of fixation support that is most widely used, it is the insoluble immobilization gel that forms by behind the calcium chloride solution calcification water soluble algae acid sodium (SA), its preparation method is simple, cheap, but the phosphoric acid salt in the substratum can make gradually calcium alginate gel break and disintegrate and short problem in work-ing life occurs, and as a crosslinked closely macromole, the combination of alginate calcium and water molecules is not very desirable, and water ratio is not very high (70.1%).In recent years, the powerful agglutinating value(of coal) that polyvinyl alcohol (PVA) is given because of its unique texture (containing a large amount of hydroxyls on the molecular chain), epithelium snappiness, smoothness, wear resistance etc. are used as immobilization embedded dose widely, but meanwhile powerful agglomeration tendency is so that mutually cohere water ratio lower (40.5%) between the particle.The researchs such as Wang Xiaohua are found to form reticulated structure behind hydrophilic polymer SA and the PVA composite crosslinking, mesh wherein makes water molecules easily enter in the polymkeric substance, water ratio increases substantially (80.6%), and the tensile yield of matrix material brings up to 400% from original 80% and 280%, and elasticity and toughness increase substantially.In addition, SiO 2As a kind of inorganics can with organic SA and PVA with chemical bonds, Lin Songbai etc. studies show that SA-SiO 2Chemical bond has made up orderly immobilized enzyme embedding environment; Moreover, SiO 2As the atom type crystal, intensity is higher, is added in the gel particle to gain in strength, and SiO is added in experimental results show that of Song Xiangyang etc. in the sodium alginate soln 2The effect of rear tolerate P-levels acid buffer is better.SA-PVA-SiO involved in the present invention 2The advantage of the above research of co-immobilization technological synthesis, for the yeast saccharomyces cerevisiae of glucose fermentation producing and ethanol ( Saccharomyces cerevisiae) and the candida tropicalis of xylose fermentation ethanol ( Candida tropicalis) made up good growth and fermentation microenvironment, so that the glucose and xylose in the rice straw enzymatic saccharification liquid can both effectively be transformed, improved the ethanol conversion of rice straw.Can realize that through co-immobilization this mixed thalline is repeatedly to be recycled and reused for Rice Straw Fermented straw saccharification liquid to produce ethanol.
Summary of the invention
The object of the present invention is to provide a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw, had the high and low energy consumption of ethanol fermentation transformation efficiency and free of contamination advantage.
The present invention is achieved like this, and its processing step comprises:
1. bacterial strain is selected: Trichodermareesei ( Trichoderma reesei), viride ( Trichoderma viride), yeast saccharomyces cerevisiae ( Saccharomyces cerevisiae), candida tropicalis ( Candida tropicalis);
2. mixed thalline is the structure of I: Trichodermareesei and viride are pressed in the 6% inoculum size access rice straw fermented substratum with the ratio of 2 ﹕ 1,34 ℃, 120 rpm shaking tables are cultivated 4 d, so that reaching maximum enzyme, cellulase lives, meanwhile Institute of Micro-biology's product lignin-degrading enzymes can be realized the preliminary pre-treatment to stalk, and the whole process of follow-up in-situ enzymolysis is followed in this pre-treatment.The prescription of rice straw fermented substratum is: rice straw powder 6 g, wheat bran 3 g, soybean cake powder 3 g, Semen Maydis powder 2 g, KH 2PO 40.5 g, CaCl 20.3 g, water 100 mL.
3. in-situ enzymolysis saccharification: behind mixed fermentation 4 d, improve mixed fungus fermentation temperature to 50 ℃, the thalline inactivation, stop the consumption to glucose, Ligninolytic Enzymes is followed cellulase hydrolysis process performance Effect of Pretreatment all the time, clear away the obstacles for cellulosic enzymolysis, Trichodermareesei and lignocellulolyticenzymes that viride produces are realized efficient enzymatic saccharification in position, continue to obtain rice straw enzymatic saccharification product behind 72 h;
4. the preparation of straw saccharification liquid: 4 layers of filtered through gauze enzymatic saccharification product, the centrifugal 25min of 4000 rpm gets 121 ℃ of sterilizations of supernatant liquor, 30 min as follow-up batch of Rice Straw Fermented saccharified liquid.Concentration of reduced sugar is 22.74g/L in the gained straw saccharification liquid, and wherein hexose content accounts for 81.71%, and pentose content accounts for 18.29%;
5. mixed thalline is the structure of II: (number of cells reaches 4.84 * 10 to get concentration and be the yeast saccharomyces cerevisiae bacteria suspension of 0.1 g/mL 8Individual/mL) 0.5 mL and concentration are that (number of cells reaches 3.43 * 10 to 0.1 g/mL Candida tropicalis suspension 8Individual/mL) embedding medium (2%SA, 6%PVA, 1%SiO of 0.5 mL and 20 mL 2Solution) mix after, change in the 20 mL syringes, splash into 100 mL3%CaCl 2, 5%H 3BO 3In the solution, obtaining diameter is the curing bead of 3~5 mm, the processing of spending the night under 4 ℃ of conditions;
6. co-immobilization mixed yeast particle batch Rice Straw Fermented saccharified liquid producing and ethanol: get above-mentioned co-immobilization bead 20 mL, inoculation enters in the 30 mL straw saccharification liquid, 28 ℃, after 90 rpm shaking tables are cultivated 4 h, fermentation liquid is toppled over out, again change the fermentation that fresh straw saccharification liquid carries out next batch.The ethanol content of fermentation liquid is 4.97~5.05 g/L, and alcohol production speed can reach 2.170 g/ (Lh);
Technique effect of the present invention is: it is low that the present invention has energy consumption, low for equipment requirements, do not bring the pollution problem of soda acid, the raw materials pretreatment cost is low, the cellulase cost is low, the mixed thalline that makes up is II energy Reusability after the co-immobilization technical finesse, thereby has the reducing sugar that efficiently utilizes in the straw saccharification liquid, the advantages such as ethanol fermentation intensity height; The present invention also has following advantage: 1. the present invention is obtaining remarkable effect aspect the Biological Pretreatment of lignocellulose.Constructed mixed thalline is that I Trichodermareesei and viride not only have complementary lignin-degrading enzymes and live among the present invention, and Mierocrystalline cellulose, hemicellulase are lived stronger.Therefore should mixed thalline be the growth that degraded cellulose, hemicellulose gained reducing sugar can help bacterial strain, laccase, lignin peroxidase secreted in the strain growth process can be realized preliminary Biological Pretreatment to rice straw again, and this Biological Pretreatment follows the whole process of enzymatic saccharification all the time, so that can highlight in position the superiority such as the low and environmental protection of this binary system pre-treatment cost in the enzymatic saccharification process.2. the present invention has realized the low cost development utilization of cellulase.Trichodermareesei and viride mixed culture, when reaching the cellulase-producing peak, by improving temperature so that microorganism deactivated, stop it to the consumption of reducing sugar, intracellular cellulase discharges fully, reaches simultaneously the suitableeest saccharification temperature of cellulase, Institute of Micro-biology's cellulase-producing is realized efficient enzymatic saccharification in position, use the microbial enzyme that extracts purifying than general Study, saved the high cost of cellulase extraction purifying, realized cost degradation.3. the present invention is constructed utilize the glucose producing and ethanol and utilize the mixed thalline of wood sugar producing and ethanol is the efficient ethanol conversion that II has realized straw saccharification liquid, so that all kinds of reducing sugars in the straw saccharification liquid can be produced alcohol utilization fully, and the co-immobilization mixed yeast can be realized the high-density culture of yeast and recycle the shortening fermentation period.
Description of drawings
Fig. 1 is the scanning electron microscope (SEM) photograph of the front straw sample structure of degraded in the embodiment of the invention.
Fig. 2 is the scanning electron microscope (SEM) photograph of the straw sample structure behind mixed fungus fermentation in the embodiment of the invention.
Fig. 3 is the scanning electron microscope (SEM) photograph of the front co-immobilization bead tangent plane of fermentation in the embodiment of the invention.
Fig. 4 is the scanning electron microscope (SEM) photograph of co-immobilization bead tangent plane after batch fermentation in the embodiment of the invention.
Fig. 5 is process flow diagram of the present invention.
Embodiment
Elaborate below by embodiment and by reference to the accompanying drawings the present invention being done:
Embodiment: according to technical process such as Fig. 5,
1. making up mixed thalline is I enzymatic saccharification rice straw
(1) preparation of somatic cells: from culture presevation inclined-plane difference picking one transfering loop Trichodermareesei ACCC32412( Trichoderma reeseiACCC32412) and viride ACCC31931( Trichoderma virideACCC31931) thalline is inoculated in respectively in the 250 mL triangular flasks that 50 mL PDA liquid nutrient mediums are housed, and 28 ℃, 120 rpm shaking culture, 24 h namely get seed liquor;
(2) mixed thalline is the structure of I: getting Trichodermareesei ACCC32412 seed liquor 2 mL, viride ACCC31931 seed liquor 1 mL, to access simultaneously initial pH be 5.5 50 mL rice straw fermented substratum (rice straw powder 6 g, wheat bran 3 g, soybean cake powder 3 g, Semen Maydis powder 2 g, KH 2PO 40.5 g, CaCl 20.3 g, water 100mL) in, place 34 ℃, mixed culture 4 d under the 120 rpm conditions.Microorganism produces lignocellulolyticenzymes in culturing process, carry out the preliminary degraded of rice straw; Can find out that by accompanying drawing 1 and accompanying drawing 2 contrasts the rice straw degraded is very abundant after this processes.
2. the in-situ enzymolysis saccharification of microbial enzyme
After mixed thalline is I enzymatic production 4 d, improve system temperature to 50 ℃, thalline Yin Gaowen and losing activity, stop the consumption to reducing sugar, intracellular enzyme is discharged fully, the hydrolysis result of institute's cellulase-producing reaches optimum regime simultaneously, Ligninolytic Enzymes is also followed the Effect of Pretreatment of cellulase hydrolysis process performance lignin degradation all the time, for cellulosic enzymolysis is cleared away the obstacles, Trichodermareesei ACCC32412 and viride lignocellulolyticenzymes that ACCC31931 produces are realized efficient enzymatic saccharification in position, continue 72 h, obtain rice straw enzymatic saccharification product.
3. the preparation of straw saccharification liquid
With 4 layers of above-mentioned enzymatic saccharification product of filtered through gauze, the centrifugal 25min of 4000 rpm, supernatant liquor is straw saccharification liquid.Concentration of reduced sugar is 22.74 g/L in the saccharified liquid, and wherein hexose content accounts for 81.71%, and pentose content accounts for 18.29%.Saccharified liquid is namely got follow-up batch of fermention medium in 121 ℃ of sterilization 30 min.
4. making up mixed thalline is II Rice Straw Fermented saccharified liquid producing and ethanol
(1) preparation of yeast cell: from culture presevation inclined-plane difference picking one transfering loop yeast saccharomyces cerevisiae CGMCC2.346 ( Saccharomyces cerevisiaeCGMCC2.346) and candida tropicalis ACCC21256( Candida tropicalisACCC21256) thalline accesses respectively in the 100 mLPDA liquid nutrient mediums, and 30 ℃, 120 rpm shaking tables are cultivated 48 h, namely get the yeast cell culture;
(2) bacteria suspension preparation and cell counting: get respectively centrifugal 25 min of above-mentioned yeast cell culture 4000 rpm, collecting precipitation is made into the bacteria suspension that concentration is 0.1 g/mL with stroke-physiological saline solution.The number of cells that counting method of blood cell is calculated yeast saccharomyces cerevisiae CGMCC2.346 bacteria suspension reaches 4.84 * 10 8Individual/mL, the number of cells of candida tropicalis ACCC21256 bacteria suspension reaches 3.43 * 10 8Individual/mL;
(3) preparation of co-immobilization mixed yeast and activation: the embedding medium (2%SA, 6%PVA, the 1%SiO that get respectively above-mentioned yeast saccharomyces cerevisiae CGMCC2.346 bacteria suspension 0.5 mL and candida tropicalis ACCC21256 bacteria suspension 0.5 mL and 20 mL 2Solution) mix after, change in the 20 mL syringes, splash into 100 mL3%CaCl 2, 5%H 3BO 3In the solution, obtaining diameter is the curing bead of 3~5 mm, after placing 4 ℃ of refrigerator overnight, with stroke-physiological saline solution washing 2~3 times, access in the 50 mL PDA liquid nutrient mediums, carry out (30 ℃ of activation culture, after 120 rpm activate 24 h, change the fresh PDA liquid nutrient medium, continue activation 24 h), it is for subsequent use to put 4 ℃ of refrigerators; The co-immobilization bead tangent plane that obtains such as accompanying drawing 3;
(4) co-immobilization mixed yeast batch Rice Straw Fermented saccharified liquid producing and ethanol: get 20 mL SA-PVA-SiO 2The co-immobilization mixed yeast accesses in the 30 mL fermention mediums, rubber plug jam-pack bottleneck, 28 ℃, 90 rpm pour out fermentation liquid after cultivating 4 h, wash bead 2~3 times with stroke-physiological saline solution, add fresh fermention medium, carry out next batch fermentation, experiment is carried out 4 batches altogether.The potassium bichromate colorimetry records that ethanol content reaches 4.97~5.05 mg/mL in the fermentation liquid, and the ethanol generating rate can reach 2.170 g/ (Lh), co-immobilization bead tangent plane such as accompanying drawing 4 after batch fermentation.

Claims (11)

1. a method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw is characterized in that the process of described method is: (1) preparation rice straw fermented substratum; (2) utilize viride and Trichodermareesei mixed fermentation straw, collaborative secretion lignocellulolytic enzymes comprises lignin peroxidase, laccase and cellulase; (3) realize the in-situ enzymolysis saccharification of rice straw by alternating temperature technique, thereby prepare straw saccharification liquid; (4) recycling SA-PVA-SiO 2The co-immobilization technology is mixed the embedding of bacterium co-immobilization with yeast saccharomyces cerevisiae and candida tropicalis, and this co-immobilization mixed yeast particle can multiple batches of Rice Straw Fermented saccharified liquid.
2. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1 is characterized in that the prescription of described rice straw fermented substratum is: rice straw powder 6 g, wheat bran 3 g, soybean cake powder 3 g, Semen Maydis powder 2 g, KH 2PO 40.5 g, CaCl 20.3 g, water 100 mL.
3. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1, it is characterized in that described mixed fermentation refers to Trichodermareesei and viride with the ratio of 2 ﹕ 1 by in the 6% inoculum size access rice straw fermented substratum, 34 ℃, 120 rpm shaking tables are cultivated 4 d, so that reaching maximum enzyme, cellulase lives, meanwhile Institute of Micro-biology's product lignin-degrading enzymes can be realized the preliminary pre-treatment to stalk, and the whole process of follow-up in-situ enzymolysis is followed in this pre-treatment.
4. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1 is characterized in that described lignin peroxidase is that Trichodermareesei is secreted.
5. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1 is characterized in that described laccase is that viride is secreted.
6. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1 is characterized in that described cellulase is that Trichodermareesei and viride are secreted jointly.
7. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1, after it is characterized in that described alternating temperature technique refers to Trichodermareesei and viride mixed fermentation 4 d, improve mixed thalline and be temperature to 50 ℃, the thalline inactivation, stop the consumption to reducing sugar, Ligninolytic Enzymes is followed cellulase hydrolysis process performance Effect of Pretreatment all the time, for cellulosic enzymolysis is cleared away the obstacles, Institute of Micro-biology produces lignocellulolyticenzymes and realizes in position efficient enzymatic saccharification, continues to obtain rice straw enzymatic saccharification product behind 72 h.
8. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1 is characterized in that described SA-PVA-SiO 2Co-immobilization mixed yeast preparation method of granules is as follows: (1) gets candida tropicalis suspension 0.5 mL that yeast saccharomyces cerevisiae suspension 0.5 mL that concentration is 0.1 g/mL and concentration is 0.1 g/mL and the embedding medium of 20 mL mixes, wherein, number of cells reaches 4.84 * 10 in the yeast saccharomyces cerevisiae suspension 8Individual/mL, number of cells reaches 3.43 * 10 in the candida tropicalis suspension 8Individual/mL, embedding medium contains 2% sodium alginate, 6% polyvinyl alcohol and 1%SiO 2Solution; (2) mixing solutions is changed in the 20 mL syringes, splash into the 3%CaCl of 100 mL 2And 5%H 3BO 3Solution in, obtaining diameter is the co-curing bead of 3~5 mm, the processing of spending the night under 4 ℃ of conditions.
9. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1, the multiple batches of fermentation that it is characterized in that described co-immobilization mixed yeast refers to get co-immobilization bead 20 mL after the activation, inoculation enters in the 30 mL straw saccharification liquid, 28 ℃, after 90 rpm shaking tables are cultivated 4 h, fermentation liquid is toppled over out, again change fresh straw saccharification liquid, the co-immobilization mixed yeast carries out next batch fermentation again, the ethanol content of fermentation liquid is 4.97~5.05 g/L, and alcohol production speed can reach 2.170 g/ (Lh).
10. a kind of method that makes up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw as claimed in claim 1 is characterized in that concentration of reduced sugar is 22.74 g/L in the described straw saccharification liquid, and wherein hexose content accounts for 81.71%, and pentose content accounts for 18.29%.
11. such as claim 1 or 10 described a kind of methods that make up mixed bacterium system Rice Straw Fermented Ethanol Production Process with Straw, the preparation method who it is characterized in that described straw saccharification liquid is: with 4 layers of described enzymatic saccharification product of filtered through gauze claim 7, centrifugal 25 min of 4000 rpm get 121 ℃ of sterilizations of supernatant liquor, 30 min.
CN2012105170029A 2012-12-06 2012-12-06 Method of constructing mixed bacteria system for fermenting straw stalks to produce ethanol Pending CN103060418A (en)

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