阅读说明：本技术 一种利用绿藻生物质发酵生产燃料乙醇的方法 (Method for producing fuel ethanol by fermenting green alga biomass ) 是由 王辉 李萍 方飞 于 2019-11-13 设计创作，主要内容包括：本发明提供了一种利用绿藻生物质发酵生产燃料乙醇的方法。具体方法为：利用绿藻作为生物质原料进行乙醇发酵的技术方法,用绿藻、不同硫酸水解浓度以及调节钠离子浓度提升乙醇产量。石莼葡萄糖含量丰富,作为发酵乙醇的原料是十分有潜力的。预处理过程中最适硫酸浓度为2％,并利用氢氧化钙调节pH值为5.5时,其最大乙醇浓度可高达30g/L,且产乙醇速率可达0.63g/(L.h),乙醇产量为37g/kg。(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.)

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.

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.