CN110643641A - Method for producing fuel ethanol by fermenting green alga biomass - Google Patents
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
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Abstract
The invention provides a method for producing fuel ethanol by fermenting green alga biomass. The specific method comprises the following steps: the technical method for performing ethanol fermentation by using green algae as a biomass raw material improves the yield of ethanol by using the green algae, different sulfuric acid hydrolysis concentrations and regulating sodium ion concentration. The ulva has rich glucose content, and is very potential as a raw material for fermenting ethanol. The optimum sulfuric acid concentration in the pretreatment process is 2%, when the pH value is adjusted to 5.5 by using calcium hydroxide, the maximum ethanol concentration can reach 30g/L, the ethanol production rate can reach 0.63g/(L.h), and the ethanol yield is 37 g/kg.
Description
Technical Field
The invention relates to a method for producing fuel ethanol by fermenting green algae biomass, belongs to the technical field of biomass energy production in biochemical industry, and particularly relates to a technical method for producing clean and renewable energy.
Background
With the increasing energy crisis and the increasing environmental pollution, clean and renewable energy sources are generally concerned by people in the background of times with increasing environmental pollution. The currently researched biomass energy sources mainly comprise fuel ethanol, butanol, biodiesel and the like. The fuel ethanol has very wide application prospect as a green energy source, and countries such as the United states, Brazil and the like have realized the application of the fuel ethanol for vehicles, thereby greatly reducing the demand on petroleum energy. The Jilin province of China is the first province of using the ethanol gasoline for vehicles in China, and the harmful substances discharged by the automobile exhaust are reduced by 30-60%. At present, the raw materials for producing fuel ethanol at home and abroad mainly comprise grain biomass such as corn, wheat and the like, the raw material cost accounts for 50 percent of the production cost of the fuel ethanol, and the production of the fuel ethanol by using the biomass raw materials can aggravate the grain crisis of the society. The method adopts cheap non-grain raw materials to solve the grain problem possibly brought by producing fuel ethanol by using grain raw materials to a certain extent. Therefore, it has become a world consensus to find the best non-edible biomass feedstock to replace food crops for producing fuel ethanol. The production of cellulosic ethanol in the United states, Europe, Canada and other countries has been on an industrial scale, and China just completed a pilot-scale project of producing 500 tons of cellulosic ethanol every year in 2006. The third generation algal fuel ethanol has its unique superiority over the first generation biofuel ethanol represented by corn ethanol and the second generation fuel ethanol represented by cellulosic ethanol.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a technical method for effectively improving the yield of ethanol in seaweed fuel.
In order to realize the purpose, the invention adopts the following technical scheme:
a method for producing fuel ethanol by fermenting green algae biomass comprises the following steps:
step 1: after ulva and enteromorpha are respectively crushed, a certain amount of green alga powder is weighed to prepare raw materials.
Step 2: and (3) sequentially adding liquefying enzyme (80IU/g) and saccharifying enzyme (40IU/g) into the raw materials in the step (1) for enzymolysis for 48 h and 36h respectively.
And step 3: and (3) carrying out acidolysis on the raw materials subjected to enzymolysis by using sulfuric acid, treating at 120 ℃ for 2h, centrifuging at 3000r for 10min, and collecting supernatant.
And 4, step 4: and adjusting the pH value of the hydrolyzed sugar solution by using calcium hydroxide. Sterilizing at 106 deg.C for 30 min.
And 5: and (3) inoculating the dry yeast into the sucrose solution, putting the dry yeast into a constant-temperature water bath for a proper time, and then moving the dry yeast to 30 ℃ for shaking for 2h by a shaking table for activation.
Step 6: after the activation of the yeast is finished, inoculating the yeast into green algae hydrolysis sugar solution according to the inoculation amount of 10% (v/v), and fermenting in a shaking table at 30 ℃. Sampling at regular time during fermentation, and filtering to obtain filtrate for determining ethanol, reducing sugar and monosaccharide contents.
And 7: 3 parallel fermentations were performed and the mean seat final results were taken.
After the ulva and the enteromorpha are respectively crushed, a certain amount of green algae powder is weighed to prepare 350g/L of raw material.
In the step 2, the treating conditions of the liquefying enzyme are 60 ℃ and the pH value is 6.0, and the treating conditions of the saccharifying enzyme are 60 ℃ and the pH value is 4.5.
In the step 3, the raw materials after enzymolysis are respectively subjected to acidolysis by using 2%, 4% and 8% sulfuric acid, and are treated for 2 hours at 120 ℃.
In the step 4, the pH value of the hydrolyzed sugar solution is adjusted to 5.5 by using calcium hydroxide.
In the step 5, 1g of dry yeast is inoculated into 100mL of 2% sucrose solution, and placed in a constant temperature water bath at 38 ℃ for reaction for 30 min.
The invention has the advantages that:
the effect of ethanol fermentation by using green algae as a biomass raw material is effectively improved;
② when the pH value is adjusted to 5.5 by calcium hydroxide, the maximum ethanol concentration can reach 30g/L, the ethanol production rate can reach 0.63g/(Lh), and the ethanol yield is 37 g/kg.
And the process of producing the biomass ethanol is easy to operate, has no secondary pollution, has lower cost and meets the requirement of actual production.
Drawings
FIG. 1 is a graph of ethanol production over time for different species of green algae fermentations of the present invention.
FIG. 2 is a graph showing the time-dependent ethanol production from various concentrations of acid hydrolysis according to the present invention.
Detailed Description
The following non-limiting examples are presented to enable those of ordinary skill in the art to more fully understand the present invention and are not intended to limit the invention in any way.
Example 1
The method for producing fuel ethanol by fermenting green algae biomass comprises the following steps:
firstly, preparing green alga hydrolysis sugar solution:
step 1: after ulva and enteromorpha are respectively crushed, a certain amount of green algae powder is weighed to prepare 350g/L raw material.
Step 2: and (2) sequentially adding liquefying enzyme (80IU/g) and saccharifying enzyme (40IU/g) into the raw materials in the step (1), and performing enzymolysis for 48 hours and 36 hours respectively under the conditions that the treating condition of the liquefying enzyme is 60 ℃ and the pH value is 6 and the treating condition of the saccharifying enzyme is 60 ℃ and the pH value is 4.5.
And step 3: performing acidolysis on the raw material after enzymolysis by using 2% sulfuric acid, treating at 120 ℃ for 2h, centrifuging at 3000r for 10min, and collecting supernatant.
And 4, step 4: the pH of the hydrolyzed sugar solution was adjusted to 5.5 with calcium hydroxide. Sterilizing at 106 deg.C for 30 min.
And 5: three parallel fermentation experiments were performed and the average was taken.
(II) fermentation:
step 1: 1g of dry yeast is taken to be put into 100ml of 2 percent sucrose solution, placed in a constant temperature water bath at 38 ℃ for 30min, and then moved to 30 ℃ for shaking table for 2h for activation.
Step 2: after the activation of the yeast is finished, inoculating the yeast into green alga hydrolysis sugar solution according to the inoculation amount of 5% (v/v), and fermenting in a shaking table at 30 ℃.
And step 3: sampling at regular time during fermentation, and filtering to obtain filtrate for determining ethanol, reducing sugar and monosaccharide contents.
Example 2
The method for producing fuel ethanol by fermenting green algae biomass comprises the following steps:
firstly, preparing green alga hydrolysis sugar solution:
step 1: after ulva and enteromorpha are respectively crushed, a certain amount of green algae powder is weighed to prepare 350g/L raw material.
Step 2: and (2) sequentially adding liquefying enzyme (80IU/g) and saccharifying enzyme (40IU/g) into the raw materials in the step (1), and performing enzymolysis for 48 hours and 36 hours respectively under the conditions that the treating condition of the liquefying enzyme is 60 ℃ and the pH value is 6 and the treating condition of the saccharifying enzyme is 60 ℃ and the pH value is 4.5.
And step 3: performing acidolysis on the raw material after enzymolysis by using 4% sulfuric acid, treating at 120 ℃ for 2h, centrifuging at 3000r for 10min, and collecting supernatant.
And 4, step 4: the pH of the hydrolyzed sugar solution was adjusted to 5.5 with calcium hydroxide. Sterilizing at 106 deg.C for 30 min.
And 5: three parallel fermentation experiments were performed and the average was taken.
(III) fermentation:
step 1: 1g of dry yeast is taken to be put into 100ml of 4 percent sucrose solution, placed in a constant temperature water bath at 38 ℃ for 30min, and then moved to 30 ℃ for shaking table for 2h for activation.
Step 2: after the activation of the yeast is finished, inoculating the yeast into green algae hydrolysis sugar solution according to the inoculation amount of 10% (v/v), and fermenting in a shaking table at 30 ℃.
And step 3: sampling at regular time during fermentation, and filtering to obtain filtrate for determining ethanol, reducing sugar and monosaccharide contents.
Example 3
The method for producing fuel ethanol by fermenting green algae biomass comprises the following steps:
firstly, preparing green alga hydrolysis sugar solution:
step 1: after ulva and enteromorpha are respectively crushed, a certain amount of green algae powder is weighed to prepare 350g/L raw material.
Step 2: and (2) sequentially adding liquefying enzyme (80IU/g) and saccharifying enzyme (40IU/g) into the raw materials in the step (1), and performing enzymolysis for 48 hours and 36 hours respectively under the conditions that the treating condition of the liquefying enzyme is 60 ℃ and the pH value is 6 and the treating condition of the saccharifying enzyme is 60 ℃ and the pH value is 4.5.
And step 3: performing acidolysis on the raw material after enzymolysis by using 8% sulfuric acid, treating at 120 ℃ for 2h, centrifuging at 3000r for 10min, and collecting supernatant.
And 4, step 4: the pH of the hydrolyzed sugar solution was adjusted to 5.5 with calcium hydroxide. Sterilizing at 106 deg.C for 30 min.
And 5: three parallel fermentation experiments were performed and the average was taken.
(IV) fermentation:
step 1: 1g of dry yeast is taken to be put into 100ml of 8 percent sucrose solution, placed in a constant temperature water bath at 38 ℃ for 30min, and then moved to 30 ℃ for shaking table for 2h for activation.
Step 2: after the activation of the yeast is finished, inoculating the yeast into green algae hydrolysis sugar solution according to the inoculation amount of 15% (v/v), and fermenting in a shaking table at 30 ℃.
And step 3: sampling at regular time during fermentation, and filtering to obtain filtrate for determining ethanol, reducing sugar and monosaccharide contents.
Example 4
The method for producing fuel ethanol by fermenting green algae biomass comprises the following steps:
firstly, preparing green alga hydrolysis sugar solution:
step 1: after ulva and enteromorpha are respectively crushed, a certain amount of green algae powder is weighed to prepare 350g/L raw material.
Step 2: and (2) sequentially adding liquefying enzyme (80IU/g) and saccharifying enzyme (40IU/g) into the raw materials in the step (1), and performing enzymolysis for 48 hours and 36 hours respectively under the conditions that the treating condition of the liquefying enzyme is 60 ℃ and the pH value is 6 and the treating condition of the saccharifying enzyme is 60 ℃ and the pH value is 4.5.
And step 3: performing acidolysis on the raw material after enzymolysis by using 2% sulfuric acid, treating at 120 ℃ for 2h, centrifuging at 3000r for 10min, and collecting supernatant.
And 4, step 4: the pH of the hydrolyzed sugar solution was adjusted to 5.5 with calcium hydroxide. Sterilizing at 106 deg.C for 30 min.
And 5: three parallel fermentation experiments were performed and the average was taken.
(V) fermentation:
step 1: 1g of dry yeast is taken to be put into 100ml of 2 percent sucrose solution, placed in a constant temperature water bath at 38 ℃ for 30min, and then moved to 30 ℃ for shaking table for 2h for activation.
Step 2: after the activation of the yeast is finished, inoculating the yeast into green algae hydrolysis sugar solution according to the inoculation amount of 10% (v/v), and fermenting in a shaking table at 30 ℃.
And step 3: sampling at regular time during fermentation, and filtering to obtain filtrate for determining ethanol, reducing sugar and monosaccharide contents.
While the invention has been described with reference to exemplary embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from the essential scope thereof. Therefore, it is intended that the invention not be limited to the particular embodiment disclosed, but that the invention will include all embodiments falling within the scope of the appended claims.
Claims (6)
1. A method for producing fuel ethanol by fermenting green algae biomass is characterized by comprising the following steps:
step 1: after ulva and enteromorpha are respectively crushed, a certain amount of green alga powder is weighed to prepare raw materials.
Step 2: and (3) sequentially adding liquefying enzyme (80IU/g) and saccharifying enzyme (40IU/g) into the raw materials in the step (1) for enzymolysis for 48 h and 36h respectively.
And step 3: and (3) carrying out acidolysis on the raw materials subjected to enzymolysis by using sulfuric acid, treating at 120 ℃ for 2h, centrifuging at 3000r for 10min, and collecting supernatant.
And 4, step 4: and adjusting the pH value of the hydrolyzed sugar solution by using calcium hydroxide. Sterilizing at 106 deg.C for 30 min.
And 5: and (3) inoculating the dry yeast into the sucrose solution, putting the dry yeast into a constant-temperature water bath for a proper time, and then moving the dry yeast to 30 ℃ for shaking for 2h by a shaking table for activation.
Step 6: after the activation of the yeast is finished, inoculating the yeast into green algae hydrolysis sugar solution according to the inoculation amount of 10% (v/v), and fermenting in a shaking table at 30 ℃. Sampling at regular time during fermentation, and filtering to obtain filtrate for determining ethanol, reducing sugar and monosaccharide contents.
And 7: 3 parallel fermentations were performed and the mean seat final results were taken.
2. The method for producing fuel ethanol by using green alga biomass fermentation according to claim 1, wherein in the step 1, after the ulva and the enteromorpha are respectively crushed, a certain amount of green alga powder is weighed to prepare raw materials of 350 g/L.
3. The method for producing fuel ethanol by fermenting chlorella biomass according to claim 1, wherein in the step 2, the liquefying enzyme treatment condition is 60 ℃ and the pH value is 6.0, and the saccharifying enzyme treatment condition is 60 ℃ and the pH value is 4.5.
4. The method for producing fuel ethanol by fermenting green algae biomass as claimed in claim 1, wherein in the step 3, the raw material after enzymolysis is respectively subjected to acidolysis by 2%, 4% and 8% sulfuric acid and treated at 120 ℃ for 2 h.
5. The method for producing fuel ethanol by fermenting green algae biomass as claimed in claim 1, wherein in the step 4, the pH value of the hydrolyzed sugar solution is adjusted to 5.5 by using calcium hydroxide.
6. The method for producing fuel ethanol by using green alga biomass fermentation as claimed in claim 1, characterized in that in the step 5, 1g of dry yeast is inoculated into 100mL of 2% sucrose solution, and the mixture is placed in a constant temperature water bath at 38 ℃ for reaction for 30 min.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BE838665A (en) * | 1976-02-18 | 1976-06-16 | METHOD FOR MANUFACTURING ETHYL ALCOHOL FROM SINGLE-CELL GREEN ALGAE | |
| CN105368881A (en) * | 2015-12-10 | 2016-03-02 | 辽宁石油化工大学 | Method for preparing fuel ethanol by joint fermentation of synanthrin containing material and cellulosic material |
| CN105420286A (en) * | 2015-11-24 | 2016-03-23 | 广西罗城科潮基业科技发展有限公司 | Alcohol production method |
| CN106834358A (en) * | 2017-03-21 | 2017-06-13 | 青岛大学 | A kind of method that Efficient Conversion algal polysaccharides prepare bio-ethanol |
-
2019
- 2019-11-13 CN CN201911103272.3A patent/CN110643641A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BE838665A (en) * | 1976-02-18 | 1976-06-16 | METHOD FOR MANUFACTURING ETHYL ALCOHOL FROM SINGLE-CELL GREEN ALGAE | |
| CN105420286A (en) * | 2015-11-24 | 2016-03-23 | 广西罗城科潮基业科技发展有限公司 | Alcohol production method |
| CN105368881A (en) * | 2015-12-10 | 2016-03-02 | 辽宁石油化工大学 | Method for preparing fuel ethanol by joint fermentation of synanthrin containing material and cellulosic material |
| CN106834358A (en) * | 2017-03-21 | 2017-06-13 | 青岛大学 | A kind of method that Efficient Conversion algal polysaccharides prepare bio-ethanol |
Non-Patent Citations (2)
| Title |
|---|
| GRAEME M WALKER等: "Enhancing Yeast Alcoholic Fermentations", 《ADVANCES IN APPLIED MICROBIOLOGY》 * |
| 王辉等: "绿藻生物质发酵生产燃料乙醇的研究", 《功能材料》 * |
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Application publication date: 20200103 |