CN101633939A - Method for promoting enzymolysis and fermentation of straw cellulose by fresh straw wall protein - Google Patents
Method for promoting enzymolysis and fermentation of straw cellulose by fresh straw wall protein Download PDFInfo
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- CN101633939A CN101633939A CN200810134420A CN200810134420A CN101633939A CN 101633939 A CN101633939 A CN 101633939A CN 200810134420 A CN200810134420 A CN 200810134420A CN 200810134420 A CN200810134420 A CN 200810134420A CN 101633939 A CN101633939 A CN 101633939A
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Abstract
The invention provides a method for promoting enzymolysis and conversion of straw lignocelluloses by fresh straw wall protein. Aiming at the characteristics of the fresh straw wall protein, the invention develops a novel process for enzymolysis and fermentation of straw, and applies the rich and cheap endogenous cellulose enzyme and cofactors thereof in the straw to straw conversion. In the novel process for the straw conversion based on fresh straw wall protein application, firstly the fresh straw wall protein is extracted high efficiently and economically, and then is mixed with the straw pretreated by steam explosion for enzymolysis and fermentation. The process not only improves the conversion efficiency of cellulose and reduces the cost of cellulase, but also saves the cost of fresh straw pretreatment link. The fresh maize straw is taken as a substrate of the enzymolysis and the fermentation, and is also taken as a source of the cellulase and the cofactors, so that the aim of promoting the conversion of the cellulose by the self resource of the straw is achieved.
Description
Technical field
The invention belongs to enzymolysis, the fermentation technical field of lignocellulose, particularly utilize cell wall protein in the fresh corn stalk the promoter action of stalk enzymolysis, fermentation, the novel straw enzymolysis of member, the novel process of fermentation.Fresh straw not only as the substrate of enzymolysis, fermentation, also as cellulase and collaborative proteic source thereof, promotes cellulosic conversion.
Background technology
Straw lignocellulose is a renewable resources the abundantest on the earth, and effectively utilizing stalk also is one of effective ways of alleviating the human problems such as food shortage, energy dilemma and environmental pollution that faced.The lignocellulosic material transforming fuel alcohol is one of important channel of Renewable Energy Development, worldwide obtains day by day paying attention to.It is the key link that transforms that the stalk enzymolysis becomes fermentable sugars, but present enzymolysis process still exists problems such as enzymolysis efficiency is low, fermentable sugars concentration is low, cellulase cost height.At present both at home and abroad aspect cellulosic ethanol, also carry out deep research, and obtained interim progress, but still be difficult to realize suitability for industrialized production extensive, economic, cleaning so far.Simultaneous saccharification and fermentation ethanol have equipment simple, save time, raise the efficiency, reduce advantage such as living contaminants, therefore be one of valid approach of present cellulose conversion ethanol.Yet inconsistent at enzymolysis, leavening temperature, still there are a lot of problems in aspects such as enzyme cost.
Lignocellulose has extremely complicated structure, natural cellulose aggregated structure complexity and degree of crystallinity height, intramolecularly and the intermolecular hydrogen bond that exists; Mierocrystalline cellulose twines mutually with macromolecular substance such as hemicellulose, xylogen simultaneously, forms fine and close three-dimensional structure.The structure of this complexity has reduced cellulosic accessibility and reactive behavior, has increased the difficulty of hydrolysis.
Except Mierocrystalline cellulose, there are relatively abundanter endogenic cellulase and synergistic factor thereof in the plant cell wall, in each stage of plant-growth, zymoprotein has participated in processes such as lax, synthetic, the decomposition of cell walls.Compare characteristics such as the effect of plant endogenous sexual cell wall zymoprotein pair cell wall has efficiently, gene regulating complexity with the cellulase in other source.Especially in fresh straw, also there is relatively abundanter cell wall protein at maize straw, comprising endogenous cellulase and synergistic factor thereof.
The cellulase of plant endogenous property and synergistic factor thereof have source advantage such as abundant, efficient, and its additive as cellulose degradation is applied to the conversion of stalk, and also cost, the raising enzymolysis efficiency that reduces cellulase for amplitude provides opportunity.The present invention starts with from the stalk cell wall protein that cellulose degradation is had promoter action, from economical, efficiently utilize the angle of plant endogenous property cellulase and synergistic factor thereof, explore the enzymolysis and the new process for fermenting of stalk.In operational path, the fresh corn stalk not only acts on the substrate of enzymolysis, also promotes cellulosic conversion as the enzyme source.The straw aboundresources, stalk cell wall protein is applied to the degraded of lignocellulose, not only improved cellulosic enzymolysis efficiency, also opened up brand-new direction for the cost that reduces cellulase, and realized the higher value application of resource having reached the purpose of utilizing plant own resource efficient degradation.
Summary of the invention
The present invention relates to the method that fresh straw wall protein promotes stalk cellulose enzymolysis, fermentation, it is applied to stalk enzymolysis, fermentation as the cellulase additive with cell wall protein abundant in the fresh straw, thereby reduce cost, the raising enzymolysis efficiency of cellulase, reach the purpose of utilizing the stalk own resource to promote cellulose degradation.
Design of the present invention is as follows: in fresh corn stalk (FCS), abundant cell wall protein is arranged still, they have participated in the adjusting that cell wall structure changes, and can promote cellulosic conversion in cellulase hydrolysis.Cell wall protein in the fresh corn stalk is extracted by simple method, then the quick-fried maize straw of mixture and vapour (SECS) is mixed, regulate concentration of substrate and pH, carry out enzymolysis and fermentation then respectively.The fresh corn stalk is not only as the substrate of enzymolysis, and endogenic cell wall protein can also promote cellulosic enzymolysis.Fresh straw and steam puffed stalk are carried out mixed fermentation, not only improved enzymolysis efficiency, reduced the cost of cellulase, also saved the preprocessing process of substrate, reduced the step of enzymolysis, fermentation.
Fresh straw wall protein promotes the method for stalk cellulose enzymolysis, fermentation, it is characterized in that may further comprise the steps:
1. the extraction of maize straw cell wall protein: will be crushed to 40-100 purpose fresh corn stalk and place the cell wall protein extracting solution, and add protein extract according to solid-to-liquid ratio 1/4-1/10,4 ℃ leave standstill the extraction that 12-24h carries out cell wall protein.
2. the mixing of fresh straw and steam puffed stalk: utilize acetic acid to regulate pH to 5.0, mix than 1/3-3/1 according to dry weight according to fresh and steam explosion maize straw, (50mM pH5.0) adjusts concentration of substrate to 20-100mg/ml to add acetate buffer solution.
3. enzymolysis: on the basis of adjusting fresh and the quick-fried maize straw ratio of vapour, concentration of substrate and pH, add cellulase according to 10-30FPU/ (g substrate) and carry out enzymolysis, temperature 45-50 ℃, time 48-72h.
4. simultaneous saccharification and fermentation ethanol: on the basis of adjusting fresh and the quick-fried maize straw ratio of vapour, concentration of substrate and pH, add cellulase, add the activation yeast according to 10-40FPU/ (g substrate), temperature 30-37 ℃, time 72-96h.
In order to analyze by the effect behind the process optimization, respectively with fresh, steam explosion maize straw, and both blended direct fermentations are analyzed ethanol production for contrast.When being substrate with fresh and steam explosion maize straw respectively, alcoholic acid output is respectively 1.50 and 1.34mg/ml.Steam explosion and fresh corn stalk according to dry weight than 1/1 (FCS ﹠amp; SECS) directly during mixed fermentation, produced the ethanol of 1.56mg/ml, fermented separately than steam explosion and fresh corn stalk respectively and improved 16.4% and 4%.And the fresh corn stalk carries out the extraction of cell wall protein earlier, then (the FCS during according to 1/1 mixed fermentation with the quick-fried maize straw of vapour; SECS (N)), alcoholic acid output is 2.85mg/ml, has improved 82.7% (table 1) than both direct mixed fermentation.
Ethanol production in the table 1 different fermentations system
The present invention has the following advantages:
1) stalk cell wall protein and cellulase have synergy, can increase substantially cellulosic transformation efficiency, save the consumption of cellulase, reach to utilize own resource to promote the purpose that transforms.
2) in conversion processes such as enzymolysis, fermentation, fresh straw is not only as substrate, and as the additive of cellulase, stalk has been saved the pre-treatment cost without preprocessing process.
3) the fresh straw source is abundant, and this had both guaranteed competent raw material supply, had realized the efficient utilization of stalk resource again.
4) optimum temps of fresh straw endogenous cellulase is consistent with yeast, has solved the inconsistent problem of enzymolysis and leavening temperature to a certain extent.
Description of drawings
Fig. 1 is an ethanol production in the different maize straw fermentation systems
Respectively take fresh and steam blasting maize straw during as substrate, enzymolysis 72h, the output of glucose is 4.87 And 5.15mg/ml; During than 1/1 direct mixed enzymolysis (FCS ﹠ SECS), glucose is given birth to according to dry weight for both Become 5.21mg/ml; And the fresh corn stalk carries out earlier the extraction of cell wall protein, then with the quick-fried corn stalk of vapour When stalk mixes according to 1/1 (FCS ﹠ SECS (N)), glucose has generated 8.3mg/ml; Respectively with fresh When maize straw and steam blasting maize straw were the substrate enzymolysis, cellobiose had produced respectively 0.76 and 3.61 Mg/ml; When both mix direct enzymolysis according to 1/1, produced 2.1mg/ml; Transform by enzymolysis process After, produced 0.45mg/ml cellobiose (Fig. 1). The fresh corn stalk cell wall protein is being extracted in explanation, During mixed enzymolysis, the effect of beta-glucosidase has obtained effective performance then.
Embodiment
Enzymolysis, fermentation efficiency that example 1 utilizes the fresh corn stalk cell wall protein to improve maize straw will be crushed to 60 purpose fresh corn stalks and place the cell wall protein extracting solution, add protein extract according to solid-to-liquid ratio 1/5,4 ℃ leave standstill the extraction that 12h carries out cell wall protein.Utilize acetic acid to regulate mixture pH to 4.8.Mix than 1/1 according to dry weight according to fresh and steam explosion maize straw, add acetate buffer solution (50mM, pH 4.8) and adjust concentration of substrate to 50mg/ml; On the basis of adjusting fresh and the quick-fried maize straw ratio of vapour, concentration of substrate and pH, add cellulase according to 20FPU/ (g substrate) and carry out enzymolysis, 45 ℃ of temperature, time 48h.On the basis of adjusting fresh and steam puffed stalk ratio, concentration of substrate and pH, add cellulase, 37 ℃ of temperature, time 72h according to 30FPU/ (g substrate).
Enzymolysis, fermentation efficiency that example 2 utilizes the fresh corn stalk cell wall protein to improve the quick-fried wheat stalk of vapour will be crushed to 40 purpose fresh corn stalks and place the cell wall protein extracting solution, add protein extract according to solid-to-liquid ratio 1/6,4 ℃ leave standstill the extraction that 15h carries out cell wall protein.Utilize acetic acid to regulate mixture pH to 4.8.Then fresh corn stalk and the quick-fried wheat stalk of vapour are mixed than 1/2 according to dry weight, add acetate buffer solution (60mM, pH 4.8) and adjust concentration of substrate to 80mg/ml; On the basis of adjusting fresh corn stalk and the quick-fried wheat stalk ratio of vapour, concentration of substrate and pH, add cellulase according to 15FPU/ (g substrate) and carry out enzymolysis, 50 ℃ of temperature, time 48h.On the basis of adjusting fresh and steam puffed stalk ratio, concentration of substrate and pH, add cellulase, 37 ℃ of temperature, time 72h according to 30FPU/ (g substrate).
Example 3 utilizes the fresh corn stalk cell wall protein to improve enzymolysis, the fermentation efficiency of the quick-fried straw of vapour
To be crushed to 80 purpose fresh corn stalks and place the cell wall protein extracting solution, and add protein extract according to solid-to-liquid ratio 1/7,4 ℃ leave standstill the extraction that 20h carries out cell wall protein.Utilize acetic acid to regulate mixture pH to 4.5.Then fresh corn stalk and the quick-fried straw of vapour are mixed than 1/3 according to dry weight, add acetate buffer solution (50mM, pH 4.5) and adjust concentration of substrate to 60mg/ml; On the basis of adjusting fresh corn stalk and ratio, concentration of substrate and pH, add cellulase according to 12FPU/ (g substrate) and carry out enzymolysis, 45 ℃ of temperature, time 48h.On the basis of adjusting fresh and steam puffed stalk ratio, concentration of substrate and pH, add cellulase, 35 ℃ of temperature, time 72h according to 25FPU/ (g substrate).
Example 4 utilizes the fresh corn stalk cell wall protein to improve enzymolysis, the fermentation efficiency of broomcorn straw
To be crushed to 100 purpose fresh corn stalks and place the cell wall protein extracting solution, and add protein extract according to solid-to-liquid ratio 1/4,4 ℃ leave standstill the extraction that 20h carries out cell wall protein.Utilize acetic acid to regulate mixture pH to 5.0.Mix than 1/3 according to dry weight according to fresh and steam explosion maize straw, add acetate buffer solution (50mM, pH 4.5) and adjust concentration of substrate to 80mg/ml; On the basis of adjusting fresh corn stalk and broomcorn straw ratio, concentration of substrate and pH, add cellulase according to 20FPU/ (g substrate) and carry out enzymolysis, 50 ℃ of temperature, time 48h.On the basis of adjusting fresh and steam puffed stalk ratio, concentration of substrate and pH, add cellulase, 37 ℃ of temperature, time 72h according to 30FPU/ (g substrate).
Example 5 utilizes the fresh corn stalk cell wall protein to improve enzymolysis, the fermentation efficiency of bambusa textile grass
To be crushed to 100 purpose fresh corn stalks and place the cell wall protein extracting solution, and add protein extract according to solid-to-liquid ratio 1/4,4 ℃ leave standstill the extraction that 20h carries out cell wall protein.Utilize acetic acid to regulate mixture pH to 5.0.Then fresh corn stalk and steam explosion bambusa textile grass are mixed than 1/3 according to dry weight, add acetate buffer solution (50mM, pH 4.5) and adjust concentration of substrate to 80mg/ml; On the basis of adjusting fresh corn stalk and bambusa textile grass ratio, concentration of substrate and pH, add cellulase according to 20FPU/ (g substrate) and carry out enzymolysis, 50 ℃ of temperature, time 48h.On the basis of adjusting fresh and steam puffed stalk ratio, concentration of substrate and pH, add cellulase, 37 ℃ of temperature, time 72h according to 30FPU/ (g substrate).
Claims (4)
1. fresh straw wall protein promotes the method for stalk cellulose enzymolysis, fermentation, it is characterized in that may further comprise the steps:
1) extraction of stalk cell wall protein: will be crushed to 40-100 purpose fresh corn stalk and place the cell wall protein extracting solution, and add protein extract according to solid-to-liquid ratio 1/4-1/10,4 ℃ leave standstill the extraction that 12-24h carries out cell wall protein;
2) mixing of fresh straw and steam puffed stalk: utilize acetic acid to regulate pH to 4.0-5.0, mix than 1/3-3/1 according to dry weight according to fresh and steam explosion maize straw, add acetate buffer solution (20-100mM, pH 4.0-5.0) and adjust concentration of substrate to 20-100mg/ml;
3) enzymolysis: on the basis of adjusting fresh straw and steam puffed stalk ratio, concentration of substrate and pH, add cellulase according to 10-30FPU/ (g substrate) and carry out enzymolysis, temperature 45-50 ℃, time 48-72h;
4) simultaneous saccharification and fermentation ethanol: on the basis of adjusting fresh and steam puffed stalk ratio, concentration of substrate and pH, add cellulase, temperature 30-37 ℃, time 72-96h according to 10-40FPU/ (g substrate).
2. method according to claim 1 is characterized in that the source of stalk cell wall protein comprises corn stalk, straw, beanstalk, sorghum stalks and wheat straw.
3. method according to claim 1, the extraction damping fluid that it is characterized in that is: 15-30mM phosphoric acid buffer, 0.5-1.5M NaCl, 0.5-2mM phenylmethylsulfonyl fluoride, pH 4.5-7.0.
4. method according to claim 1 is characterized in that stalk comprises maize straw, straw, wheat straw, sorghum stalks, beanstalk, Herba Poae Sphondylodis and cogongrass.
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| CN2008101344203A CN101633939B (en) | 2008-07-23 | 2008-07-23 | Method for promoting enzymolysis and fermentation of straw cellulose by fresh straw wall protein |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104814407A (en) * | 2015-05-18 | 2015-08-05 | 青岛海之星生物科技有限公司 | Nutrition powder based on sweet potatoes and preparation method of nutrition powder |
| CN106036019A (en) * | 2016-06-25 | 2016-10-26 | 陈毅忠 | Method for preparing compound feed sweetener by utilizing corn stalks |
| CN111454995A (en) * | 2020-04-08 | 2020-07-28 | 华中农业大学 | Method for promoting biomass material to produce alcohol |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1806945B (en) * | 2005-01-20 | 2010-05-12 | 中国科学院过程工程研究所 | Method for complete enzymolysis of straw cellulose by utilizing pretreatment and enzymolysis process |
| CN101092447B (en) * | 2006-06-23 | 2010-08-04 | 中国科学院过程工程研究所 | Fibrin enzymolysis auxiliary agent from stalk stuff of plant |
| US20080131947A1 (en) * | 2006-12-01 | 2008-06-05 | Cellencor, Inc. | Treatment of Cellulosic Material for Ethanol Production |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104814407A (en) * | 2015-05-18 | 2015-08-05 | 青岛海之星生物科技有限公司 | Nutrition powder based on sweet potatoes and preparation method of nutrition powder |
| CN106036019A (en) * | 2016-06-25 | 2016-10-26 | 陈毅忠 | Method for preparing compound feed sweetener by utilizing corn stalks |
| CN111454995A (en) * | 2020-04-08 | 2020-07-28 | 华中农业大学 | Method for promoting biomass material to produce alcohol |
| CN111454995B (en) * | 2020-04-08 | 2021-09-24 | 华中农业大学 | Method for promoting biomass material to produce alcohol |
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