CN102994572A - Method for producing butanol by fermentation of cellulose in bamboos - Google Patents
Method for producing butanol by fermentation of cellulose in bamboos Download PDFInfo
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- CN102994572A CN102994572A CN2012105094881A CN201210509488A CN102994572A CN 102994572 A CN102994572 A CN 102994572A CN 2012105094881 A CN2012105094881 A CN 2012105094881A CN 201210509488 A CN201210509488 A CN 201210509488A CN 102994572 A CN102994572 A CN 102994572A
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Abstract
The invention relates to the field of biological energy sources, and in particular relates to a method for producing butanol by fermentation of cellulose in bamboos. The method comprises the following steps of: 1) pre-processing; 2) solid-liquid separation; 3) enzymolysis; 4) preparation of a fermentation culture medium; 5) anaerobic fermentation; and 6) rectification of a fermented product, so that the butanol is obtained. By adopting the glucose obtained by hydrolysis of cellulose rich in bamboos to serve as a carbon source and be used for growth and fermentation of biobutanol strains, a new path is developed for utilization of bamboo resources, the recycling of processing wastes of the bamboos also can be realized, and a cheap raw material is provided for fermenting the butanol at the same time.
Description
Technical field
The present invention relates to the bioenergy field, be specifically related to a kind of method of utilizing the cellulose fermentation in the bamboo to produce butanols.
Background technology
Butanols originally is the large basic material that serves many purposes, in recent years studies show that butanols also is a kind of high-grade fuel and fuel dope, its high boiling point (118 ℃) and low-steam pressure help the cold start-up of automobile; Because the hydrophobicity of butanols is stronger than ethanol, therefore be easier to vapour, diesel hydrocarbon class A fuel A miscible; The perfect combustion of butanols can reduce the CO in the tail gas greatly
2Discharging, and the residual hydrocarbons pollution does not occur, very favourable to purifying air.Above-mentioned advantage can make butanols become alternate-engine novel green fuel, one of renewable energy resources of the Sustainable development of alternative mineralising fuel.According to statistics, global butanols apparent consumption amount was estimated at 3,500,000 tons in 2012, about 1,200,000 tons of the butanols consumption of China, degree of self-sufficiency less than 50%.
Along with oil price rise steadily and to the impact of Economic development, the research and development of accelerating biomass energy and not relying on petroleum base chemicals production method have become day by day urgent problem.Therefore, fermentative Production butanols technology has caused widely concern again as a kind of important tachnical storage.Past fermentative Production butanols is take grain raw material (such as corn, wheat etc.) as substrate, higher provision price causes expense of raw materials to account for the ratio higher (more than 60%) of solvent total production cost, this has not only limited the butanols competitiveness of product in market, also runs counter to the grain security strategy of China.Also just for these reasons, National Development and Reform Committee will have suspended putting on record of corn deep processing project and has checked and approved the end of the year 2007.Therefore, making butanols take lignocellulose as raw materials through biotransformation is the inexorable trend of Future Development.
China is topmost product bamboo state in the world, is that kind, area, accumulation and the bamboo wood of bamboo, the output of bamboo shoots all rank first place, the world.The bamboo kind is known to have 39 to belong to more than 500 kinds, about 5,000,000 hectares of the bamboo grove total area, and wherein mao bamboo woods have 3,000,000 hectares, and annual bamboo wood ultimate production reaches 1,200,000,000.China's bamboo resource mainly is distributed in the ground such as Sichuan, Zhejiang, Jiangxi, Anhui, Hunan, Hubei, Fujian, Guangdong, Guangxi, Guizhou, Chongqing, Yunnan, wherein maximum with Fujian (1,000,000 hectares), Jiangxi (700,000 hectares), Hunan (700,000 hectares) and Zhejiang (640,000 hectares) 4 provinces, account for more than 60% of the national bamboo grove total area.Every year can be cut bamboo wood more than 1000 ten thousand steres by China, be equivalent to about 1/5 of year Wood cut, also can produce a large amount of bamboo branches in this external bamboo processing process, ring, the bamboo spring, bamboo bits the like waste, its quantity accounts for more than 60% of former bamboo weight at least, bamboo lignocellulose rich content, wherein Mierocrystalline cellulose accounts for 35%(w/w), hemicellulose level accounts for 20%(w/w), can obtain containing the hydrolyzed solution of glucose and xylose mixture after the hydrolysis, and can be used for growth and the fermentation of biological butanol bacterial strain fully, be the desirable alternative materials of past grain fermentation butyl alcohol.Utilize bamboo fermentative production butanols, the new approach that not only has been the bamboo resource exploitation can also realize the recycling of bamboo processing waste providing a kind of more cheap raw material for butylic fermentation simultaneously.Utilize at present bamboo to produce alcohol fuel and appear in the newspapers, but have no the research that utilizes bamboo to produce butanols.
Summary of the invention
The object of the invention is to overcome the defective of prior art, a kind of method of utilizing the cellulose fermentation in the bamboo to produce butanols is provided, utilize the bamboo resource of China's abundant to be raw material production biofuel butanols, realize the recycling of bamboo processing waste, and reduce the production cost of butanols.
The present invention at first discloses a kind of method of utilizing the cellulose fermentation in the bamboo to produce butanols, and concrete steps are as follows:
1) pre-treatment: bamboo mixes the bamboo powder after pulverizing and obtaining the bamboo powder with water or dilute sulphuric acid, react 10～30min under 170～200 ℃ of temperature, obtains the pre-treatment product;
2) solid-liquid separation: with step 1) pre-treatment product solid-liquid separation, the solid residue of getting in the pre-treatment product is repeatedly washed;
3) enzymolysis: with step 2) solid residue after the washing mixes by certain solid-to-liquid ratio with water, obtains the enzymolysis mixture, after the pH value of adjusting enzymolysis mixture is 4.5～5.5, adds cellulase and beta-glucosidase and carries out enzyme digestion reaction, obtains enzymolysis product;
4) fermention medium preparation: with the enzymolysis product solid-liquid separation of step 3), get enzymolysis solution and add nitrogenous source preparation fermention medium;
5) utilize the fermention medium anaerobic condition bottom fermentation after sterilizing to cultivate the production of butanol bacterial strain, obtain tunning;
6) butanols is collected in tunning rectifying.
More excellent, the described pulverizing of step 1) is for grinding or the ultra micro comminution by gas stream.
More excellent, the particle diameter of the described bamboo powder of step 1) is 10～2000 orders; More excellent, the particle diameter of the described bamboo powder of step 1) is 1500～2000 orders.
More excellent, the concentration of the described dilute sulphuric acid of step 1) is less than or equal to 2wt%.Dilute sulphuric acid of the present invention is the aqueous solution of sulfuric acid, and wherein the mass percent of sulfuric acid greater than 0, less than or equal to 2.
More excellent, the solid-to-liquid ratio that the described bamboo powder of step 1) mixes with pretreatment fluid is 1:6～1:12, and is excellent in selecting 1:6～1:10.
More excellent, step 1) pretreatment reaction temperature condition is 170～180 ℃.
More excellent, step 2) and the method for the described solid-liquid separation of step 4) can be centrifugal or filter.
By the control of pretreatment condition, make the hemicellulose degraded in the bamboo, the solidliquid mixture that pre-treatment obtains carries out solid-liquid separation through centrifugal or filtration, and the liquid main component is the degraded product wood sugar of hemicellulose, can be used for fermentation methane; The solid main component is Mierocrystalline cellulose and xylogen, is used for follow-up enzymolysis after the washing.
More excellent, step 3) enzymolysis step, the solid-to-liquid ratio of described solid residue and water are 1:6～1:10.
More excellent, the consumption of cellulase described in the step 3) is the 10-30FPU/g substrate; The consumption of beta-glucosidase is the 1-10CBU/g substrate.
More excellent, the condition of the described enzyme digestion reaction of step 3) is: hydrolysis temperature is 45～55 ℃, and enzymolysis time is 45～60h, and shaking speed is 120～180rpm.
More excellent, the condition of the described enzyme digestion reaction of step 3) is: hydrolysis temperature is 50 ℃, and the time is 48h, and rotating speed is 150r/min.
The solidliquid mixture that enzymolysis obtains carries out solid-liquid separation through centrifugal or filtration, and the liquid main component is cellulosic degraded product glucose, is used for next step fermentation butyl alcohol; The solid main component is xylogen, can be used for producing wood plastic composite etc.
More excellent, the described nitrogenous source of step 4) is corn steep liquor, ammonium sulfate, ammonium acetate or ammonium chloride one.
More excellent, the addition of the described nitrogenous source of step 4) is 0.1～1w/v%
Access production of butanol bacterial strain behind 105 ℃ of sterilizations of the substratum of preparation 15min.
More excellent, the described production of butanol bacterial strain of step 5) is selected from CICC8016, CICC8008, NCIMB8052 or ATCC55025.
More excellent, the condition of the described anaerobically fermenting of step 5) is 35～38 ℃, fermentation time is 40～50h.
The present invention utilizes the Mierocrystalline cellulose of rich content in the bamboo, it is carbon source that its hydrolysis is obtained glucose afterwards, and can be used for growth and the fermentation of biological butanol bacterial strain fully, it is the desirable alternative materials of past grain fermentation butyl alcohol, the new approach that not only has been the bamboo resource exploitation, can also realize the recycling of bamboo processing waste, provide a kind of more cheap raw material for butylic fermentation simultaneously.
Description of drawings
Fig. 1: the production scheme that utilizes the cellulose fermentation production butanols in the bamboo
Embodiment
Further set forth the present invention below in conjunction with specific embodiment, should be understood that embodiment only is used for explanation the present invention and is not used in restriction protection scope of the present invention.
Notice, the not concrete processing unit that indicates or device all adopt conventional equipment or the device in this area in the following example; Should be understood that in addition that one or more method stepss of mentioning among the present invention do not repel before and after described combination step can also exist the additive method step or can also insert the additive method step between these steps of clearly mentioning.And, except as otherwise noted, the numbering of various method steps is only for differentiating the convenient tool of various method steps, but not ordering or the enforceable scope of restriction the present invention for limiting various method steps, the change of its relativeness or adjustment, in the situation without essence change technology contents, when also being considered as the enforceable category of the present invention.
Embodiment 1
To bamboo (Jiangxi mao bamboon, Ph.pubescens) carry out the ultra micro comminution by gas stream, taking by weighing through ultra micro comminution by gas stream to particle diameter is 2000 purpose bamboo powder 5g, add 50mL, sulphuric acid soln 0.5%(w/w) (solid-to-liquid ratio is 1:10), 180 ℃ of reaction 20min, pre-treatment hydrolysis liquid suction filtration, solid residue after the washing adds 30mL water and mixes (solid-to-liquid ratio is 1:6), regulatory enzyme enzymatic hydrolysis system pH value is 4.8, add cellulase (available from Novozymes Company) 20FPU/g substrate, beta-glucosidase (available from Novozymes Company) 8CBU/g substrate, 50 ℃ of enzymolysis 48h, shaking speed is 150r/min.The enzymolysis solution suction filtration, the filtrate of getting behind the enzymolysis adds 1.0% (w/v) corn starch, connects the CICC8016 bacterial strain, and 37 ℃ of anaerobism are cultivated 48h and are obtained tunnings.In the tunning, butanols 11.98g/L, total solvent 20.13g/L.
Embodiment 2
Taking by weighing through ultra micro comminution by gas stream to particle diameter is 1500 purpose bamboo powder 5g, add 40mL, 1%(w/w) sulphuric acid soln (solid-to-liquid ratio is 1:8), 170 ℃ of reaction 20min, pre-treatment hydrolysis liquid suction filtration, solid residue after the washing adds 40mL water and mixes (solid-to-liquid ratio is 1:7), and regulatory enzyme enzymatic hydrolysis system pH value is 5.0, adds cellulase 30FPU/g substrate, beta-glucosidase 10CBU/g substrate, 45 ℃ of enzymolysis 45h, shaking speed is 120r/min.The enzymolysis solution suction filtration, clear liquid adds 0.3% (w/v) ammonium sulfate, connects the CICC8008 bacterial strain, and 38 ℃ of anaerobism are cultivated 40h, obtain tunning.Butanols 10.73g/L in the tunning, total solvent 19.04g/L.
Embodiment 3
Taking by weighing through Ordinary pulverization to particle diameter is 100 purpose bamboo powder 5g, adding the 60mL(solid-to-liquid ratio is 1:12), 0.3%(w/w) sulphuric acid soln, 200 ℃ of reaction 10min, the centrifugal 10min of pre-treatment hydrolysis liquid 5000r/min, solid residue after the washing adds 50mL water and mixes (solid-to-liquid ratio is 1:10), and regulatory enzyme enzymatic hydrolysis system pH value is 5.5, adds cellulase 20FPU/g substrate, beta-glucosidase 4CBU/g substrate, 50 ℃ of enzymolysis 60h, shaking speed is 180r/min.The centrifugal 10min of enzymolysis solution 5000r/min, clear liquid adds 0.5% (w/v) ammonium chloride, connects the NCIMB8052 bacterial strain, and 35 ℃ of anaerobism are cultivated 50h, obtain tunning.Butanols 7.75g/L in the tunning, total solvent 13.66g/L.
Embodiment 4
Taking by weighing through ultra micro comminution by gas stream to particle diameter is 2000 purpose bamboo powder 5g, add 30mL, 0.5%(w/w) sulphuric acid soln (solid-to-liquid ratio is 1:6), 180 ℃ of reaction 30min, pre-treatment hydrolysis liquid suction filtration, solid residue after the washing adds 40mL water and mixes (solid-to-liquid ratio is 1:8), and regulatory enzyme enzymatic hydrolysis system pH value is 4.5, adds cellulase 25FPU/g substrate, beta-glucosidase 2CBU/g substrate, 55 ℃ of enzymolysis 48h, shaking speed is 150r/min.Enzymolysis solution suction filtration, clear liquid add 0.5% corn steep liquor, connect the NCIMB8052 bacterial strain, and 38 ℃ of anaerobism are cultivated 40h.Butanols 9.18g/L in the tunning, total solvent 11.69g/L.
Embodiment 5
Taking by weighing through Ordinary pulverization to particle diameter is 10 purpose bamboo powder 5g, add 50mL, 2.0%(w/w) sulphuric acid soln (solid-to-liquid ratio is 1:10), 180 ℃ of reaction 20min, pre-treatment hydrolysis liquid suction filtration, solid residue after the washing adds 50mL water and mixes (solid-to-liquid ratio is 1:10), and regulatory enzyme enzymatic hydrolysis system pH value is 5.5, adds cellulase 10FPU/g substrate, beta-glucosidase 1CBU/g substrate, 50 ℃ of enzymolysis 48h, shaking speed is 150r/min.Enzymolysis solution suction filtration, clear liquid add 0.1% corn steep liquor, connect the ATCC55025 bacterial strain, and 38 ℃ of anaerobism are cultivated 45h.Butanols 6.56g/L in the tunning, total solvent 9.33g/L.
Embodiment 6
Taking by weighing through ultra micro comminution by gas stream to particle diameter is 1500 purpose bamboo powder 5g, add 50mL water (solid-to-liquid ratio is 1:10), 190 ℃ of reaction 20min, pre-treatment hydrolysis liquid suction filtration, solid residue after the washing adds 30mL water and mixes (solid-to-liquid ratio is 1:6), and regulatory enzyme enzymatic hydrolysis system pH value is 4.8, adds cellulase 15FPU/g substrate, beta-glucosidase 8CBU/g substrate, 50 ℃ of enzymolysis 48h, shaking speed is 120r/min.The enzymolysis solution suction filtration, the filtrate of getting behind the enzymolysis adds 0.5% (w/v) corn starch, connects the CICC8016 bacterial strain, and 37 ℃ of anaerobism are cultivated 48h.In the tunning, butanols 5.68g/L, total solvent 8.12g/L.
More than show and described ultimate principle of the present invention, principal character and advantage of the present invention.The technician of the industry should understand; the present invention is not restricted to the described embodiments; that describes in above-described embodiment and the specification sheets just illustrates principle of the present invention; the present invention also has various changes and modifications without departing from the spirit and scope of the present invention, and these changes and improvements all fall in the claimed scope of the invention.The claimed scope of the present invention is defined by appending claims and equivalent thereof.
Claims (10)
1. method of utilizing the cellulose fermentation in the bamboo to produce butanols, concrete steps are as follows:
1) pre-treatment: bamboo mixes the bamboo powder after pulverizing and obtaining the bamboo powder with water or dilute sulphuric acid, react 10～30min under 170～200 ℃ of temperature, obtains the pre-treatment product;
2) solid-liquid separation: with step 1) pre-treatment product solid-liquid separation, the solid residue of getting in the pre-treatment product is repeatedly washed;
3) enzymolysis: with step 2) solid residue after the washing mixes by certain solid-to-liquid ratio with water, obtains the enzymolysis mixture, after the pH value of adjusting enzymolysis mixture is 4.5～5.5, adds cellulase and beta-glucosidase and carries out enzyme digestion reaction, obtains enzymolysis product;
4) fermention medium preparation: with the enzymolysis product solid-liquid separation of step 3), get enzymolysis solution and add nitrogenous source preparation fermention medium;
5) utilize the fermention medium anaerobic condition bottom fermentation after sterilizing to cultivate the production of butanol bacterial strain, obtain tunning;
6) butanols is collected in tunning rectifying.
2. the method for claim 1 is characterized in that, the particle diameter of the described bamboo powder of step 1) is 10～2000 orders.
3. the method for claim 1 is characterized in that, the concentration of the described dilute sulphuric acid of step 1) is less than or equal to 2wt%.
4. the method for claim 1 is characterized in that, the solid-to-liquid ratio that the described bamboo powder of step 1) mixes with pretreatment fluid is 1:6～1:12.
5. the method for claim 1 is characterized in that, in the step 3) enzymolysis process, the solid-to-liquid ratio of described solid residue and water is 1:6～1:10.
6. the method for claim 1 is characterized in that, the consumption of cellulase described in the step 3) is 10～30FPU/g substrate; The consumption of beta-glucosidase is 1～10CBU/g substrate.
7. the method for claim 1 is characterized in that, the condition of the described enzyme digestion reaction of step 3) is: hydrolysis temperature is 45～55 ℃, and enzymolysis time is 45～60h, and shaking speed is 120～180rpm.
8. the method for claim 1 is characterized in that, the described nitrogenous source of step 4) is corn steep liquor, ammonium sulfate, ammonium acetate or ammonium chloride.
9. method as claimed in claim 8 is characterized in that, the addition of the described nitrogenous source of step 4) is 0.1～1w/v%.
10. the method for claim 1 is characterized in that, the described production of butanol bacterial strain of step 5) is selected from CICC8016, CICC8008, NCIMB8052 or ATCC55025.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104561190A (en) * | 2014-12-26 | 2015-04-29 | 中国林业科学研究院林产化学工业研究所 | Two-step pretreatment method for producing fermentable sugars through enzyme hydrolysis of bamboo biomass wastes |
| CN105603004A (en) * | 2016-01-22 | 2016-05-25 | 中国水产科学研究院黄海水产研究所 | Method for preparing biological butyl alcohol with marine algae as raw materials |
| CN111154814A (en) * | 2019-11-15 | 2020-05-15 | 陕西山河生物科技有限公司 | Process method for green production of gamma-polyglutamic acid from bamboo sugar solution |
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| CN102260230A (en) * | 2011-05-17 | 2011-11-30 | 中国林业科学研究院林产化学工业研究所 | Method for preparing furfural by using two-step hydrolysis of biomass |
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| CN102260230A (en) * | 2011-05-17 | 2011-11-30 | 中国林业科学研究院林产化学工业研究所 | Method for preparing furfural by using two-step hydrolysis of biomass |
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| 金强: "生物质半纤维素稀酸水解反应", 《化学进展》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104561190A (en) * | 2014-12-26 | 2015-04-29 | 中国林业科学研究院林产化学工业研究所 | Two-step pretreatment method for producing fermentable sugars through enzyme hydrolysis of bamboo biomass wastes |
| CN104561190B (en) * | 2014-12-26 | 2018-07-27 | 中国林业科学研究院林产化学工业研究所 | A kind of two-step method method of utilizing bamboo biomass waste enzyme hydrolysis production fermentable sugars |
| CN105603004A (en) * | 2016-01-22 | 2016-05-25 | 中国水产科学研究院黄海水产研究所 | Method for preparing biological butyl alcohol with marine algae as raw materials |
| CN111154814A (en) * | 2019-11-15 | 2020-05-15 | 陕西山河生物科技有限公司 | Process method for green production of gamma-polyglutamic acid from bamboo sugar solution |
| CN111154814B (en) * | 2019-11-15 | 2023-06-02 | 陕西山河生物科技有限公司 | Technological method for green production of gamma-polyglutamic acid from bamboo sugar solution |
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