CN100375781C - Production of biological diesel oil by fixed enzyme method - Google Patents
Production of biological diesel oil by fixed enzyme method Download PDFInfo
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- CN100375781C CN100375781C CNB2005101009663A CN200510100966A CN100375781C CN 100375781 C CN100375781 C CN 100375781C CN B2005101009663 A CNB2005101009663 A CN B2005101009663A CN 200510100966 A CN200510100966 A CN 200510100966A CN 100375781 C CN100375781 C CN 100375781C
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- Y02E50/10—Biofuels, e.g. bio-diesel
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Abstract
The present invention relates to a method using a fixed lipase method to produce biological diesel oil through carrying out catalyzed alcoholysis for the greases of animals and plants, which comprises a lipase fixing step using a fixative composed of sodium alginate, kaolin, glutaraldehyde, gelatin and lactose, a step that spherical fixed enzymes are stacked in reactors filled with bed enzyme columns in a natural state, a step that the greases of animals and plants are carried out with alcoholysis under the condition of using the biological diesel oil as solvent, and a biological diesel oil producing step. The production method of the present invention has the advantages of mild reaction condition, low energy consumption, continuous production and low production cost, and meanwhile, the present invention uses biocatalyst so as to have strong reaction specificity and few byproducts; the produced biological diesel oil has the characteristics of light color and high quality.
Description
Technical field
This relates to the production method of bio-diesel oil, and especially a kind of immobilized lipase enzyme process is a method of bio-diesel oil at non-aqueous system catalyzed alcoholysis animal-plant oil.
Background technology
Diesel oil is the major impetus fuel of many oversize vehicles such as truck and diesel locomotive and generator etc., it is big that it has power, advantages of cheap price, and China's diesel oil is in great demand, the growth rate of demand of diesel oil has reached more than 6%, far above the rate of increase (3.2%) of gasoline.
The subject matter of diesel fuel applications is that efficiency of combustion is lower, and is serious to atmospheric pollution.Be tail gas pollution problem that solves fuel oil and the environmental stress that goes from bad to worse, people begin one's study and adopt other energy such as fuel alcohol fuel gasoline replacing, fuel alcohol is at Bei Meizhou at present, occupied significant proportion with South America as Brazil, Argentina etc. as the U.S., Canada etc., the automobile that is equipped with the fuel alcohol engine is put on market.But fuel alcohol is only as the quid pro quo of gasoline, and engine needs to change.It is the big power car of fuel that great majority are needed diesel oil, and fuel alcohol also is not suitable for.And the tail gas pollution that causes of diesel oil is than big many of gasoline, the therefore surrogate-biofuel of necessary exploitation diesel oil.
There are abundant Vegetable oil lipoprotein and animal grease resource in China, and the annual soya-bean oil annual production of China reaches 6,000 ten thousand tons, and a large amount of sewer oil and the frying oils of restaurant's generation, and food-processing industry also produces a large amount of tankage.If in addition fine utilization can solve troubling " sewer oil " and wait problem of environmental pollution, provide green fuel.
Biofuel all is to use chemical method production at present, promptly uses animal, Vegetable oil lipoprotein and low-carbon alcohol such as methyl alcohol or ethanol under basic catalyst, under high temperature (230 ℃-250 ℃), and transesterification generation methyl esters or ethyl ester.But the chemical method biodiesel synthesis has following shortcoming: complex process, and alcohol must be excessive, and follow-up must have corresponding pure retrieving arrangement; The energy consumption height, color and luster is dark, because unsaturated fatty acids is at high temperature apt to deteriorate in the fat, esterification products is difficult to reclaim the cost height; Generative process has the waste lye discharging.
For addressing the above problem, people begin one's study and use the biological enzyme biodiesel synthesis, and promptly animal-plant oil and low-carbon alcohol are carried out transesterification by lipase, prepare corresponding fatty acid methyl ester and ethyl ester.The enzyme process biodiesel synthesis has mild condition, pure consumption is little, advantages such as non-pollutant discharge.But at present because low-carbon alcohol is toxic to enzyme, cause the enzymatic conversion method rate very low, and enzyme deactivation is serious, enzyme and enzyme immobilization cost are higher, and above variety of problems makes the Production by Enzymes biofuel still can not industrialization.
Method of the present invention promptly is the problem that faces at process for preparing biodiesel by enzyme, replace conventional solvent such as expensive normal hexane, sherwood oil with biofuel, and used novel, cheap process for fixation enzyme is carried out immobilization, can effectively reduce the cost of process for preparing biodiesel by enzyme.
Summary of the invention
What the purpose of this invention is to provide that a kind of cost is low, productivity is high utilizes sodium alginate immobilized enzyme production method of bio-diesel oil, and this method can be utilized fixed lipase catalyzed conversion waste animal vegetable oil production biofuel.
For reaching above-mentioned purpose, production method of the present invention comprises the steps:
A. lipase immobilization: sodium alginate soln, kaolin solution, glutaraldehyde solution, gelatin solution, lactose solution are mixed, again mixed solution is placed on the magnetic stirring apparatus, add an amount of lipase, stirring makes and mixes, and puts into 30 ℃ of-50 ℃ of constant temperature shaking tables then, and 100-300r/min is vibration 1-2h down, splash into then in the calcium chloride solution of 15-30g/L, form spheroidal particle, solidify being fixed of suction filtration enzyme;
B. immobilized enzyme alcoholysis animal-plant oil: immobilized enzyme is piled up in first and second and the three reactor with state of nature; Be 1: 1 mixed in molar ratio with animal-plant oil and methyl alcohol, and adding and grease weight ratio are 4: 1-5: 1 biofuel is formed substrate mixture as the water of solvent and grease weight 15%-25%, this substrate mixture places the substrate mixing tank and carries out the magnetic agitation heating, so that methyl alcohol and vegetable and animals oils mix; Substrate mixes after volume pump extracts to enter in the first step reactor reacts under immobilized enzyme catalysis, the reaction solution that comes out from first step reactor is through after the glycerin separator separation of glycerin and enter second stage reactor after 1: 1 the methanol mixed of mol ratio and react, the reaction solution that comes out from second stage reactor through glycerine separate and and 1: 1 methanol mixed of mol ratio after enter third stage reactor and react, the final reaction product can make biofuel after the glycerine separation.
In the preparation method's of the present invention steps A, during immobilized lipase, the concentration of sodium alginate soln is 20-40g/L, the concentration of kaolin solution is 10-30g/L, the weight percentage of glutaraldehyde solution is 2.5-8%, the concentration of gelatin solution is 10-20g/L, and the concentration of lactose solution is 5-20g/L; The concentration that its preferred concentration is sodium alginate soln is 30g/L, the concentration of kaolin solution is 20g/L, and the weight percentage of glutaraldehyde solution is 5%, and the concentration of gelatin solution is 10g/L, the concentration of lactose solution is 5g/L, and each liquor capacity is 300: 300: 5 than optimum: 5: 3.
Solidification value described in preparation method's steps A of biofuel of the present invention is 3-5 ℃, and the set time is 4-6h.
Calcium chloride (CaCl
2) concentration range of solution is 15-30g/L, preferred concentration is 20g/L.
The inventive method is applicable to various types of lipase, and the mass ratio of lipase and sodium alginate is 1: 1-1: 4, be preferably 3: 5.
Compare with preparation biofuel in the prior art, the present invention has the following stated advantage:
(1) do the immobilized enzyme good springiness that carrier is made with sodium alginate, change to such an extent that tolerance is more intense, be difficult for being dissolved in water temperature, pH value, the physical strength height, enzyme is lived stable, and the transformation period is longer.Sodium alginate itself is nontoxic, and cheap, material is easy to get, and is as safe as a house during therefore as the carrier of immobilized enzyme, and process for fixation is fairly simple, and the condition during immobilization is relatively gentleer, and is very few to the enzymic activity influence.
(2) the common fixing agent of forming by sodium alginate, kaolin, glutaraldehyde, gelatin, lactose, improved the hydrophilic and hydrophobic property of fixation support effectively, for the effect between enzyme and the substrate provides good microenvironment, thereby improved the catalytic activity of enzyme greatly.
(3) replace expensive normal hexane, sherwood oil to make solvent with biofuel, can effectively reduce the cost of process for preparing biodiesel by enzyme.
(4) present method is reactor used is packed bed enzyme column type reactor, and the ball shape fixation enzyme is piled up in the reactor with state of nature, and after reaction finished, immobilized enzyme also was easy to separate with water-retaining agent with reaction solution, helped reclaiming multiplexing and serialization production.
Put it briefly, compare with traditional chemical synthesis, reaction conditions gentleness of the present invention, energy consumption is low, and can serialization production, greatly reduces production cost.The present invention simultaneously uses biological catalyst, and atopic is strong, and by product is few, and prepared biofuel has lighter color, characteristics that quality is high.
Description of drawings
Fig. 1 is the device synoptic diagram of the inventive method.
Description of reference numerals: 1 is the methyl alcohol storage tank; 2 is the substrate mixing tank; 3 is volume pump; 4 is mixing tank; 5,6,7 is respectively first, second and third stage reactor; 8,9,10th, glycerin separator; The 11st, the glycerine collector.
Embodiment
As shown in Figure 1, reactor 5 and 6, be filled with spheric sodium alginate immobilized enzyme in 7, this immobilized enzyme is 1 for the mass ratio of adding and sodium alginate in sodium alginate soln, kaolin solution, glutaraldehyde solution, gelatin solution, lactose solution mix: 1-1: 4 lipase, putting into the constant temperature shaking table after mixing vibrates, dropwise splash in the calcium chloride solution of 15-30g/L then, usually, in the 100ml calcium chloride solution, splash into this mixture of the lipase that contains 0.5 gram, form spheroidal particle, 3-5 ℃ fixedly makes after 4-6h hour.Used lipase is alkaline lipase or other alkaline lipase that the Shenzhen Leveking Biology Engineering Co.,Ltd produces.
Animal-plant oil mixed with methyl alcohol, biofuel and water etc. in substrate mixing tank 2 after volume pump 3 enter in the first step reactor 5 with reactor in be filled with and react under the effect of spheric sodium alginate immobilized enzyme catalysis, the reaction solution that comes out from first step reactor is after glycerin separator 8 separation of glycerin, enter mixing tank 4 through volume pump again, in mixing tank 4 and after the methanol mixed, enter 6 reactions of second stage reactor.Methyl alcohol is squeezed into mixing tank by methyl alcohol storage tank 1 through volume pump.The reaction solution that comes out from reactor 6 enters last step reactor 7 internal reactions with methanol mixed equally after glycerine separates.The product that comes out from reactor 7 can make biofuel after glycerine separates.From glycerin separator 8,9,10 glycerine of separating enter glycerine collector 11.
Be used for realizing the inventive system comprises:
3 packed bed immobilized enzyme column type reactors 5,6 and 7, this reactor is a telescoping water-bath temperature control, makes temperature of reaction be stabilized between 40-50 ℃.
2 methyl alcohol storage tanks 1 are used for squeezing into quantitative methyl alcohol in the mixing tank 4 of reactor 6,7, realize that scalable stream adds methyl alcohol, to reduce the toxicity of low-carbon alcohol to immobilized enzyme.
1 substrate mixing tank 2 is used for stock oil, reactant, solvent etc. are mixed, and mixing tank 2 carries out the constant temperature stirring heating, so that methyl alcohol and raw material wet goods thorough mixing.
2 mixing tanks 4, the methyl alcohol and the reaction solution that are used for classification is dosed mix.
3 glycerin separator 8,9,10, the glycerine that is used for reaction process separates, and to reduce the toxicity of glycerine to enzyme, will separate as the glycerine of reaction product simultaneously, can promote the direction of reacting to generating biofuel to move, and helps improving transformation efficiency.
1 glycerine collector 11 is used to collect glycerin separator 8,9,10 isolated glycerine.
Embodiment 1:
0.5g lipase is joined in the 30ml solution that contains 0.3g sodium alginate (0.046mo1/L) and 0.6g kaolin (20g/L), the glutaraldehyde solution that adds 0.5ml gelatin (10g/L), 0.5ml lactose (10g/L) and 0.3ml 5%, stirred 10 minutes, mix, under 42 ℃ of conditions, vibration 1h injects the 100ml CaCl of 25g/L then
24 ℃ solidify 4h in the solution.Be piled up in first and second and the three reactor with state of nature;
Be 1: 1 mixed in molar ratio with animal-plant oil and methyl alcohol, and adding and grease weight ratio are that 5: 1 biofuel is formed substrate mixture as the water of solvent and grease weight 20%6, place the substrate mixing tank to carry out magnetic agitation and heating this substrate mixture, so that methyl alcohol and animal and plant grease mix; Substrate mixes after oil pump extracts to enter in the first step reactor reacts under immobilized enzyme catalysis, the reaction solution that comes out from first step reactor through glycerine separate and and 1: 1 methyl alcohol of mol ratio enter in the second stage reactor 6 after mixing tank, mixing and react, the reaction solution that comes out from second stage reactor 6 through glycerine separate and and 1: 1 methyl alcohol of mol ratio enter third stage reactor 7 after mixing tank, mixing and react, the final reaction product can make biofuel after glycerin separator 10 separation.The biofuel of being produced has lighter color, characteristics that quality is high, and its property analysis sees Table 1.
The biofuel analytical results that table 1. present embodiment is produced
| Project name | Analytical data | Unit |
| Density (15 ℃) | 0.8798 | g/cm3 |
| Viscosity (40 ℃) | 4.639 | mm2/s |
| Flash-point (cup of remaining silent) | >160 | ℃ |
| Cold filter clogging temperature | 1 | ℃ |
| S content | 4 | ppm |
| Carboloy residue | <0.05 | mass％ |
| Cetane value | 75.2 | |
| Water-content | 290 | ppm |
| Copper corrosion (50 ℃, 3h) | 1 | Level |
| Total acid value | 0.07 | mgKOH/g |
| Methanol content | 0.05 | mass％ |
| Methyl esters content | 99.68 | mass％ |
| Content of monoglyceride | Do not detect | mass％ |
| Two sweet ester contents | Do not detect | mass％ |
| Triglyceride content | Do not detect | mass％ |
| Free glycerol content | Do not detect | mass％ |
| Total glycerol content | Do not detect | mass％ |
| P content | 4.1 | ppm |
| Calorific value | 36.95 | kJ/g |
Embodiment 2:
The 0.5g enzyme is joined in the 30ml solution that contains 0.6g sodium alginate (0.046mol/L) and 0.6g kaolin (20g/L), the glutaraldehyde solution that adds 0.5ml gelatin (10g/L), 0.5ml lactose (10g/L) and 0.3ml 5%, stirred 10 minutes, mix, under 40 ℃ of conditions, vibration 1h injects the 100mlCaCl of 20g/L then
24 ℃ solidify 4h in the solution, filter, and are piled up in first and second and the three reactor with state of nature.
Be 1: 1 mixed in molar ratio with animal-plant oil and methyl alcohol, and adding and grease weight ratio are that 5: 1 biofuel is formed substrate mixture as the water of solvent and grease weight 20%, this substrate mixture is placed the substrate mixing tank and carry out the magnetic agitation heating, so that methyl alcohol and animal-plant oil mix; Substrate mixture mixes after oil pump extracts to enter in the first step reactor reacts under immobilized enzyme catalysis, and all the other steps are with embodiment 1 preparation biofuel.The biofuel property analysis of being produced sees Table 2.
The biofuel analytical results that table 2. present embodiment is produced
| Project name | Analytical data | Unit |
| Density (15 ℃) | 0.8816 | g/cm3 |
| Viscosity (40 ℃) | 4.647 | mm2/s |
| Flash-point (cup of remaining silent) | >160 | ℃ |
| Cold filter clogging temperature | 1 | ℃ |
| S content | 4 | ppm |
| Carboloy residue | <0.04 | mass％ |
| Cetane value | 75.2 | |
| Water-content | 290 | ppm |
| Copper corrosion (50 ℃, 3h) | 1 | Level |
| Total acid value | 0.07 | mgKOH/g |
| Methanol content | 0.05 | masss％ |
| Methyl esters content | 99.68 | mass％ |
| Content of monoglyceride | Do not detect | mass％ |
| Two sweet ester contents | Do not detect | mass％ |
| Triglyceride content | Do not detect | mass％ |
| Free glycerol content | Do not detect | mass％ |
| Total glycerol content | Do not detect | mass％ |
| P content | 4.1 | ppm |
| Calorific value | 36.95 | 1kJ/g |
Embodiment 3:
The 0.5g enzyme is joined in the 30ml solution that contains 0.9g sodium alginate (0.046mol/L) and 0.6g kaolin (20g/L), the glutaraldehyde solution that adds 0.5ml gelatin (10g/L), 0.5ml lactose (5g/L) and 0.3ml 5%, stirred 10 minutes, mix, under 42 ℃ of conditions, vibration 1h injects the 100mlCaCl of 20g/L then
24 ℃ solidify 4h in the solution, are piled up in first and second and the three reactor with state of nature.
Be 1: 1 mixed in molar ratio with animal-plant oil and methyl alcohol, and adding and grease weight ratio are that 4: 1 biofuel weighs 20% water as solvent and grease and forms substrate mixture, this substrate mixture is placed the substrate mixing tank and carry out the magnetic agitation heating, so that methyl alcohol and animal-plant oil mix; Substrate mixture mixes after oil pump extracts to enter in the first step reactor reacts under immobilized enzyme catalysis, and all the other steps are with embodiment 1 preparation biofuel.The biofuel property analysis of being produced sees Table 3.
The biofuel analytical results that table 3. present embodiment is produced
| Project name | Analytical data | Unit |
| Density (15 ℃) | 0.8818 | g/cm3 |
| Viscosity (40 ℃) | 4.645 | mm2/s |
| Flash-point (cup of remaining silent) | >160 | ℃ |
| Cold filter clogging temperature | 1 | ℃ |
| S content | 4 | ppm |
| Carboloy residue | <0.04 | mass％ |
| Cetane value | 75.2 | |
| Water-content | 290 | ppm |
| Copper corrosion (50 ℃, 3h) | 1 | Level |
| Total acid value | 0.07 | mgKOH/g |
| Methanol content | 0.05 | mass％ |
| Methyl esters content | 99.68 | mass％ |
| Content of monoglyceride | Do not detect | mass％ |
| Two sweet ester contents | Do not detect | mass％ |
| Triglyceride content | Do not detect | mass％ |
| Free glycerol content | Do not detect | mass％ |
| Total glycerol content | Do not detect | mass％ |
| P content | 4.1 | ppm |
| Calorific value | 36.83 | kJ/g |
Embodiment 4:
The 0.5g enzyme is joined in the 30ml solution that contains 1.2g sodium alginate (0.046mo1/L) and 0.6g kaolin (20g/L), the glutaraldehyde solution that adds 0.5ml gelatin (10g/L), 0.5ml lactose (5g/L) and 0.3ml (5%), stirred 10 minutes, mix, under 42 ℃ of conditions, vibration 1h injects the 100ml CaCl of 20g/L then
24 ℃ solidify 4h in the solution, are piled up in first and second and the three reactor with state of nature.
Be 1: 1 mixed in molar ratio with animal-plant oil and methyl alcohol, and adding and grease weight ratio are that 4: 1 biofuel is formed substrate mixture as the water of solvent and grease 25%, this substrate mixture is placed the substrate mixing tank and carry out the magnetic agitation heating, so that methyl alcohol and animal-plant oil mix; Substrate mixture mixes after oil pump extracts to enter in the first step reactor reacts under immobilized enzyme catalysis, and all the other steps are with embodiment 1 preparation biofuel.The biofuel property analysis of being produced sees Table 4.
The biofuel analytical results that table 4. present embodiment is produced
| Project name | Analytical data | Unit |
| Density (15 ℃) | 0.8816 | g/cm3 |
| Viscosity (40 ℃) | 4.639 | mm2/s |
| Flash-point (cup of remaining silent) | >160 | ℃ |
| Cold filter clogging temperature | 1 | ℃ |
| S content | 4 | ppm |
| Carboloy residue | <0.04 | mass％ |
| Cetane value | 75.2 | |
| Water-content | 290 | ppm |
| Copper corrosion (50 ℃, 3h) | 1 | Level |
| Total acid value | 0.07 | mgKOH/g |
| Methanol content | 0.05 | mass％ |
| Methyl esters content | 99.68 | mass％ |
| Content of monoglyceride | Do not detect | mass％ |
| Two sweet ester contents | Do not detect | mass％ |
| Triglyceride content | Do not detect | mass％ |
| Free glycerol content | Do not detect | mass％ |
| Total glycerol content | Do not detect | mass％ |
| P content | 4.2 | ppm |
| Calorific value | 37.25 | kJ/g |
Claims (4)
1. production method of bio-diesel oil: it is characterized in that may further comprise the steps:
A. lipase immobilization: sodium alginate soln, kaolin solution, glutaraldehyde solution, gelatin solution, lactose solution are mixed, again mixed solution is placed on the magnetic stirring apparatus, adding is 1 with the sodium alginate mass ratio: 1-1: 4 lipase, stir, and mix, put into 30-50 ℃ of constant temperature shaking table then, 100-300r/min is vibration 1-2h down, splashes into then in the calcium chloride solution, forms spheroidal particle, solidify being fixed of suction filtration enzyme;
B. immobilized enzyme alcoholysis animal-plant oil: immobilized enzyme is piled up in first and second and the three reactor with state of nature; Be 1: 1 mixed in molar ratio with animal-plant oil and methyl alcohol, and adding and grease weight ratio are 4: 1-5: 1 biofuel is formed substrate mixture as the water of solvent and grease weight 15-25%, this substrate mixture is placed the substrate mixing tank and carry out the magnetic agitation heating, so that methyl alcohol and animal-plant oil mix; Substrate mixes after volume pump extracts to enter in the first step reactor reacts under immobilized enzyme catalysis, the reaction solution that comes out from first step reactor through after the glycerin separator separation of glycerin and methyl alcohol enter second stage reactor after mixing in 1: 1 in molar ratio and react, the reaction solution that comes out from second stage reactor through glycerine separate and and methyl alcohol mixing in 1: 1 in molar ratio after enter third stage reactor and react, the final reaction product can make biofuel after glycerine separates
Wherein, the concentration of sodium alginate soln is 20-40g/L in the steps A, the concentration of kaolin solution is 10-30g/L, the weight percentage of glutaraldehyde solution is 2.5-8%, the concentration of gelatin solution is 10-20g/L, the concentration of lactose solution is 5-20g/L, and aforementioned each liquor capacity ratio is followed successively by: 300: 300: 5: 5: 3; The concentration of described calcium chloride is 15-30g/L; Solidification value described in the steps A is 3-5 ℃, and the set time is 4-6 hour.
2. production method of bio-diesel oil according to claim 1, the concentration that it is characterized in that sodium alginate soln in the steps A is 30g/L, the concentration of kaolin solution is 20g/L, the weight percentage of glutaraldehyde solution is 5%, the concentration of gelatin solution is 10g/L, the concentration of lactose solution is 5g/L, and aforementioned each liquor capacity ratio is followed successively by: 300: 300: 5: 5: 3.
3. production method of bio-diesel oil according to claim 1, the concentration that it is characterized in that the calcium chloride described in the steps A is 20g/L.
4. production method of bio-diesel oil according to claim 1 is characterized in that the lipase that adds in the steps A and the mass ratio of sodium alginate are 3: 5.
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| CN101235351B (en) * | 2008-02-27 | 2012-07-18 | 浙江大学 | Fixed enzyme membrane reactor, preparation thereof and method for producing biological diesel oil by the same |
| CN101781398B (en) * | 2009-01-21 | 2012-05-30 | 华东理工大学 | Enzyme method for continuously producing poly(Epsilon-caprolactone) |
| CN102827825B (en) * | 2012-08-28 | 2015-06-10 | 连云港正丰生物能源有限公司 | Immobilized lipase catalyst for preparing biodiesel and application of immobilized lipase catalyst |
| CN202989098U (en) * | 2012-12-31 | 2013-06-12 | 南通大学 | Biodiesel preparation device |
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| CN1640991A (en) * | 2004-12-06 | 2005-07-20 | 华中科技大学 | Method for producing biological diesel using lipase |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103275814A (en) * | 2013-06-07 | 2013-09-04 | 华东理工大学 | Method for preparing biodiesel without by-product of glycerin by utilization of high acid value waste oil |
| CN103275814B (en) * | 2013-06-07 | 2015-02-18 | 华东理工大学 | Method for preparing biodiesel without by-product of glycerin by utilization of high acid value waste oil |
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| CN1818026A (en) | 2006-08-16 |
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