CN106554845B - Method for extracting microalgae grease - Google Patents
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Abstract
The invention discloses a method for extracting microalgae oil, which comprises the steps of (1) adding a certain amount of inorganic salt solution and alkali solution into collected microalgae, uniformly mixing, placing in a microwave generator for radiation heating, and dissolving and crushing cell walls by utilizing the synergistic effect of the alkali, the salt and the microwave; (2) the fatty acid ester released from the cells continuously has saponification reaction with alkali to generate fatty acid salt; (3) filtering to remove algae cell debris; (4) continuously adding inorganic salt solution into the system to separate out fatty acid salt, and filtering and collecting solid; (5) adding inorganic acid into the fatty acid salt, acidifying to obtain free fatty acid, and collecting the product fatty acid. The invention adopts the aqueous phase method to extract the microalgae grease, and has the advantages of simple operation process, high yield of the microalgae grease, environmental friendliness and the like.
Description
Technical Field
The invention relates to a method for extracting microalgae oil, in particular to a method for extracting microalgae oil by adopting a water phase method.
Background
Microalgae are a diverse and extremely widespread group of lower plants that grow in water. The organism has high photosynthetic reaction system, and can pass through CO2The fixation of (1) converts light energy into chemical energy and stores the chemical energy in the form of organic matters such as grease or starch in body cells. With the increasing pressure of shortage of human social resources and the increasing environmental problem, the development of biodiesel and partial fossil energy substitute products by using microalgae has become a hot spot of current research.
The production of biodiesel by using microalgae is a complex system engineering, and covers a plurality of technical links, including screening and cultivation of microalgae species, scale cultivation of microalgae, induction of oil production, collection and processing of oil and the like. The research of microalgae as a biodiesel raw material begins in the 60 th 20 th century, and in recent years, along with the development of biotechnology, microalgae resources with high oil production capacity are obtained through the biological modification of algae species, so that the novel biodiesel production mode has a very good application prospect.
The advantages of microalgae as a new biomass energy source are obvious, but in terms of utilization, from the current research progress, the microalgae are still in the starting stage. The current research mainly focuses on screening dominant algae species, improving the growth strength of oil algae and increasing the content of oil, but the research on the aspects of improving the yield of oil, establishing an industrialized extraction process method and the like is still little. However, research on microalgae aims to solve the problem of alternative energy in the field of petrochemical industry, and therefore research on efficient oil extraction methods suitable for large-scale production and application should be a research direction for a period of time in the future.
The oil content of microalgae is very different, the oil content of some microalgae which is easy to culture in large scale generally accounts for 20-50% of the dry weight of cells and is far lower than that of conventional oil crops, and therefore, some classical oil extraction methods are not suitable for extracting the oil of the microalgae. CN200810240949.3 discloses a method for simultaneously extracting oil and protein from microalgae, which takes wet algae mud as a raw material, adjusts the pH value to be alkaline or alkalescent, and carries out wall breaking of microalgae cells and dissolution of the oil and the protein through steam. Filtering the obtained microalgae slurry to remove cell residues to obtain a mixture of oil and protein, and performing oil-water separation by using a hydrocyclone to obtain microalgae oil. Because the specific surface area of microalgae cells is large and the content of phospholipid components on cell membranes is high, the method has simple preparation process, but increases the loss rate of oil components due to the adoption of a filtration pretreatment mode. CN200910060589.3 discloses a method for extracting microbial oil and short-chain alcohol fatty acid ester thereof, which comprises the following steps: adjusting the moisture content: so as to obtain a wet culture containing the grease-containing microorganisms and having a water content of 20-90%; microwave treatment: carrying out microwave radiation cell breaking on the wet culture and reducing the water content to 5-40%; short-chain alcohol treatment: partially alcoholyzing the oil in the microorganism under the action of an alkaline catalyst, and simultaneously extracting the oil; and (3) recovering the solvent: solid-liquid separation is carried out, and the mixture of the microbial oil and the short-chain alcohol fatty acid ester is obtained after the short-chain alcohol is recovered by evaporation. The method is characterized in that the microalgae is subjected to microwave treatment and cell breaking, and then is added with short-chain alcohol solvent for treatment, although the short-chain alcohol solvent has a certain dissolving effect on cell membrane phospholipid layers after cell breaking, the alcohol dissolving effect is limited, and the short-chain alcohol mainly participates in alkali catalytic reaction.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for extracting microalgae grease. The invention adopts the aqueous phase method to extract the microalgae grease, and has the advantages of simple operation process, high yield of the microalgae grease, environmental friendliness and the like.
The method for extracting the microalgae grease comprises the following steps:
(1) adding a certain amount of inorganic salt solution and alkali solution into the collected microalgae, uniformly mixing, placing in a microwave generator for radiation heating, and dissolving and crushing cell walls by utilizing the synergistic action of the alkali, the salt and the microwave;
(2) the fatty acid ester released from the cells continuously has saponification reaction with alkali to generate fatty acid salt;
(3) filtering to remove algae cell debris;
(4) continuously adding inorganic salt solution into the system to separate out fatty acid salt, and filtering and collecting solid;
(5) adding inorganic acid into the fatty acid salt, acidifying to obtain free fatty acid, and collecting the product fatty acid.
The microalgae can be any oil-producing microalgae with the capacity of accumulating grease and fatty acid, such as green algae, diatom, red algae and the like, and especially chlorella or botryococcus. The collected microalgae can be microalgae powder or microalgae mud.
The inorganic salt in the step (1) may be NaNO3、Na2CO3、NaHCO3One or more of sodium phosphate, potassium phosphate and the like, and the concentration is 0.01-0.03 mol/L. The inorganic salt solution can be prepared by using a culture solution which is separated from algae cells after the microalgae are collected. The alkali solutionCan be NaOH solution or KOH solution with the mass concentration of 10 to 30 percent. The mass-to-volume ratio (g: ml) of the collected microalgae (by mass based on dry weight) to the inorganic salt solution and the alkali solution is 1: 5-1: 10, wherein the volume ratio of the inorganic salt solution to the alkali solution is 5: 1-1: 1.
The power of the microwave generator in the step (1) is 5-20 kW, the microwave frequency can be selected from 2450MHz or 915MHz, the radiation heating temperature is 50-70 ℃, and the heating time is 15-60 min.
The algae cell debris removed in step (3) can be membrane filtration or centrifugal filtration.
The inorganic salt in the step (4) is consistent with that added in the step (1), and can be an unsaturated solution or a saturated solution with the concentration of 0.8-3 mol/L, preferably an inorganic salt saturated solution, and the volume-to-mass ratio (ml: g) of the added amount to the collected microalgae (the mass is calculated by dry weight) is 10: 1-5: 1.
The inorganic acid in the step (5) can be one or more of sulfuric acid, hydrochloric acid, nitric acid and the like, and is acidified to pH 1-4. The fatty acid can be collected by a centrifugal method, the fatty acid is better separated from the water phase after centrifugation, and an upper phase product is collected.
Compared with the prior art, the method has the following advantages:
(1) the microalgae cells are crushed by adopting a mode of combining a salt solution and an alkali solution, on one hand, the addition of the salt solution has an autolysis effect on the microalgae cells, on the other hand, the saponification effect of the alkali can quickly dissolve the cell walls of the microalgae, and the synergistic effect of the salt and the alkali can obviously improve the crushing effect of the microalgae cells.
(2) The microwave radiation is adopted to heat the mixed system of the microalgae, the salt solution and the alkali solution, and the microwave absorption capacity of the system is enhanced due to the existence of the salt and the alkali, so that compared with the traditional heating mode, the cell can be broken more thoroughly, the reaction time is shortened while the oil yield is improved, and the energy utilization efficiency is improved.
(3) Through the process of firstly converting the grease into salt and then precipitating, the grease is well separated from other water-soluble macromolecular organic matters and other impurities in algae cells, the extraction and purification of the grease are carried out in one step, and the load of pretreatment during the preparation of the biodiesel is greatly reduced.
(4) The inorganic salt used in the salting-out process is a nutrient salt in the microalgae culture medium, and the liquid separated after salting-out is rich in inorganic nutrient salt and can be continuously recycled as a component of the microalgae culture medium.
The method adopts a water phase method to extract the microalgae grease, belongs to a low-energy-consumption and environment-friendly technical route, and has the characteristics of simple process, easy amplification and high grease yield.
Detailed Description
The following examples are given to illustrate specific processes and effects of the present invention, but are not limited to the following examples.
In the present invention, the oil yield = (mass of product fatty acid/initial algal cell dry weight) × 100%. The unit of mass in the mass-to-volume ratio or the volume-to-mass ratio is g, and the unit of volume is ml.
Example 1
The microalgae is Chlorella vulgaris (Chlorella vulgaris), and is cultured in 10L photobioreactor for 10 days, and the cell concentration of the microalgae is 1.82g/L (dry weight) when the culture is finished. Centrifuging 5L of microalgae liquid, controlling the centrifugal revolution at 4000rpm, and centrifuging for 3min to obtain microalgae mud. Adding 0.03mol/L Na into the microalgae mud2CO345ml of solution and 45ml of 10% sodium hydroxide solution. After mixing uniformly, the system is placed in a microwave generator, the power of the microwave generator is set to be 5kW, the frequency of the microwave is 2450MHz, the heating temperature is 50 ℃, and the heating is carried out for 20min to carry out cell disruption and saponification reaction of the grease. Then using a membrane with a pore diameter of 10-1Filtering with micron microfiltration membrane to remove algae cell and debris. Adding 1mol/L Na into the liquid system2CO3Separating out fatty acid salt by using 90ml of solution, and filtering and collecting solid; dissolving the fatty acid salt solid in a small amount of water, adding a hydrochloric acid solution, and acidifying to pH2.0 to obtain free fatty acid; the system was centrifuged and the upper phase was collected to give 3.17g of product fatty acid with a yield of oil of 34.8%.
Example 2
The microalgae is Botryococcus, and is cultured in 10L photobioreactor for 10 days, and the cell concentration of the microalgae is 1.78g/L (dry weight) after the culture is finished. Centrifuging 5L of microalgae liquid, controlling the centrifugal revolution at 4000rpm, centrifuging for 5min to obtain microalgae mud, drying the microalgae mud, and grinding to obtain 8.9g of microalgae powder. Adding 0.02mol/L NaNO into microalgae powder358ml of solution and 29ml of 30% sodium hydroxide solution. After mixing uniformly, the system is placed in a microwave generator, the power of the microwave generator is set to be 10kW, the frequency of the microwave is 915MHz, the heating temperature is 55 ℃, and the system is heated for 30min to carry out cell disruption and saponification reaction of the grease. Then using a membrane with a pore diameter of 10-1Filtering with micron microfiltration membrane to remove algae cell and debris. Adding 1.5mol/L NaNO into the liquid system3Separating out fatty acid salt by using 45ml of solution, and filtering and collecting solid; dissolving the fatty acid salt solid in a small amount of water, adding a hydrochloric acid solution, and acidifying until the pH value is 3.0 to obtain free fatty acid; the system was centrifuged and the upper phase was collected to give 3.34g of product fatty acid with a fat yield of 37.5%.
Example 3
The microalgae is Botryococcus, and is cultured in 10L photobioreactor for 10 days, and the cell concentration of the microalgae is 1.92g/L (dry weight) after the culture is finished. Centrifuging 5L of microalgae liquid, controlling the centrifugal revolution at 4000rpm, centrifuging for 5min to obtain microalgae mud, drying the microalgae mud, and grinding to obtain 9.6g of microalgae powder. Adding KH 0.03mol/L into microalgae powder2PO440ml of solution and 40ml of 10% potassium hydroxide solution. After mixing uniformly, the system is placed in a microwave generator, the power of the microwave generator is set to be 15kW, the frequency of the microwave is 915MHz, the heating temperature is 65 ℃, and the heating is carried out for 40min, so that cell disruption and saponification reaction of the grease are carried out. Then using a membrane with a pore diameter of 10-1Filtering with micron microfiltration membrane to remove algae cell and debris. Adding KH 1.5mol/L into the liquid system2PO4Precipitating fatty acid salt with 48ml of solution, and filtering and collecting solid; dissolving the fatty acid salt solid in a small amount of water, adding a hydrochloric acid solution, and acidifying until the pH value is 3.0 to obtain free fatty acid; the system was centrifuged and the upper phase was collected to give 3.70g of product fatty acid with a fat yield of 38.5%.
Example 4
The microalgae is Chlorella, and is cultured in 10L photobioreactor for 10 days, and the cell concentration of the microalgae is 1.93g/L (dry weight) when the culture is finished. Centrifuging 5L of microalgae liquid, controlling the centrifugal revolution at 4000rpm, centrifuging for 5min to obtain microalgae mud, drying the microalgae mud, and grinding to obtain 9.65g of microalgae powder. Adding 0.01mol/L K into microalgae powder2HPO4•3H250ml of O solution and 20ml of 20% potassium hydroxide solution.
After mixing uniformly, the system is placed in a microwave generator, the power of the microwave generator is set to be 20kW, the frequency of the microwave is 915MHz, the heating temperature is 70 ℃, and the heating is carried out for 50min, so that cell disruption and saponification reaction of the grease are carried out. Then using a membrane with a pore diameter of 10-1Filtering with micron microfiltration membrane to remove algae cell and debris. Adding 1mol/L K into the liquid system2HPO4••3H260ml of O solution is used for separating out fatty acid salt, and solid is collected by filtration; dissolving the fatty acid salt solid in a small amount of water, adding a hydrochloric acid solution, and acidifying to pH4.0 to obtain free fatty acid; the system was centrifuged and the upper phase was collected to give 3.52g of product fatty acid with a yield of oil of 36.5%.
Example 5
The microalgae chlorella is cultured for 10 days in a 10L photobioreactor, and the cell concentration of the chlorella is 1.82g/L (dry weight) after the culture is finished. Centrifuging 5L of microalgae liquid, controlling the centrifugal rotation number to be 4000rpm, centrifuging for 4min to obtain microalgae mud, drying the microalgae mud, and grinding to obtain 9.1g of microalgae powder. Adding 0.02mol/L NaH into microalgae powder2PO460ml of solution and 20ml of 25% potassium hydroxide solution. After mixing uniformly, the system is placed in a microwave generator, the power of the microwave generator is set to be 20kW, the frequency of the microwave is 2450MHz, the heating temperature is 60 ℃, and the heating is carried out for 60min, so that the cell disruption and the saponification reaction of the oil are carried out. Then using a membrane with a pore diameter of 10-1Filtering with micron microfiltration membrane to remove algae cell and debris. Adding 3mol/L NaH into the liquid system2PO4Precipitating fatty acid salt with 50ml of solution, and filtering and collecting solid; dissolving the fatty acid salt solid in a small amount of water, adding hydrochloric acidAcidifying the solution to pH2.5 to obtain free fatty acid; the system was centrifuged and the upper phase was collected to give 3.59g of product fatty acid with a yield of oil 39.4%.
Example 6
The microalgae are diatom, and are cultured in a 10L photobioreactor for 10 days, and the cell concentration of the microalgae is 1.66g/L (dry weight) when the culture is finished. Centrifuging 5L of microalgae liquid, controlling the rotation speed of the centrifuge to 4000rpm, and centrifuging for 3min to obtain 8.3g of microalgae mud. Adding 0.03mol/L Na into the microalgae mud2CO340ml of solution and 10ml of 10% sodium hydroxide solution. After mixing uniformly, the system is placed in a microwave generator, the power of the microwave generator is set to be 18kW, the frequency of the microwave is 2450MHz, the heating temperature is 55 ℃, and the heating is carried out for 45min, so that the cell disruption and the saponification reaction of the oil are carried out. Then using a membrane with a pore diameter of 10-1Filtering with micron microfiltration membrane to remove algae cell and debris. Adding 1.5mol/L Na into the liquid system2CO3The solution (83 ml) was used to precipitate the fatty acid salt, and the solid was collected by filtration; dissolving the fatty acid salt solid in a small amount of water, adding a hydrochloric acid solution, and acidifying to pH2.0 to obtain free fatty acid; the system was centrifuged and the upper phase was collected to give 2.89g of product fatty acid with a yield of oil of 34.8%.
Example 7
The microalgae is Botryococcus, and is cultured in 10L photobioreactor for 10 days, and the cell concentration of the microalgae is 2.08g/L (dry weight) after the culture is finished. Centrifuging 5L of microalgae liquid, controlling the centrifugal revolution at 4000rpm, and centrifuging for 3min to obtain microalgae mud. Adding 0.02mol/L NaHCO into microalgae mud375ml of solution and 25ml of 15% sodium hydroxide solution. After mixing uniformly, the system is placed in a microwave generator, the power of the microwave generator is set to be 14kW, the frequency of the microwave is 2450MHz, the heating temperature is 45 ℃, and the heating is carried out for 55min to carry out cell disruption and saponification reaction of the oil. Then using a membrane with a pore diameter of 10-1Filtering with micron microfiltration membrane to remove algae cell and debris. Adding NaHCO of 0.8mol/L into the liquid system3Precipitating fatty acid salt with 100ml of solution, and filtering and collecting solid; dissolving the fatty acid salt solid in a small amount of water, adding a hydrochloric acid solution, and acidifying to pH2.0 to obtain free fatty acid; the body is putThe resulting mixture was centrifuged, and the upper phase was collected to obtain 3.33g of a product fatty acid, which was found to have a yield of fats and oils of 32.0%.
Example 8
Adding Na into a liquid system after membrane filtration2CO3The saturated solution (90 ml) was used to precipitate the fatty acid salt. The other conditions were the same as in example 1. 3.50g of the product fatty acid was obtained, and the yield of the oil was 38.5%.
Example 9
Adding NaNO into a liquid system after membrane filtration3The saturated solution (45 ml) was used to precipitate the fatty acid salt. The other conditions were the same as in example 2. 3.40g of the product fatty acid was obtained, and the yield of the oil was 38.7%.
Comparative example 1
In the process of crushing microalgae cells, after adding a salt solution and an alkali solution, heating the system to 50 ℃ without microwave radiation heating, stirring simultaneously, and keeping for 20min, the rest is the same as in example 1. 2.74g of fatty acid was obtained, and the yield of the oil was 30.1%.
Comparative example 2
In the process of crushing the microalgae cells, only 90ml of 10% sodium hydroxide solution is added to the microalgae mud, and no salt solution is added, the rest is the same as in example 1. 2.38g of fatty acid was obtained, and the yield of the oil was 26.2%.
Comparative example 3
In the same way as example 1, but in the process of crushing the microalgae cells, only 0.03mol/L of Na is added to the microalgae mud2CO3Mixing 90ml solution without sodium hydroxide solution, placing the above system in a microwave generator with microwave generator power of 5kW, microwave frequency of 2450MHz, heating temperature of 50 deg.C, heating for 20min, filtering with a microfiltration membrane with membrane pore diameter of 10-1 μm, and removing algae cell and debris. A hydrochloric acid solution was added to the system to acidify to pH2.0, the system was centrifuged, and the upper phase was collected. Since a part of the fatty acid ester can be hydrolyzed to generate fatty acid under acidic condition, but the hydrolysis is not complete, the upper phase is a mixed phase of the fatty acid ester and the fatty acid, which is not separated, and the fatty acid can not be extracted.
Claims (11)
1. A method for extracting microalgae grease is characterized by comprising the following steps:
(1) adding a certain amount of inorganic salt solution and alkali solution into the collected microalgae, uniformly mixing, placing in a microwave generator for radiation heating, and dissolving and crushing cell walls by utilizing the synergistic action of the alkali, the salt and the microwave; the inorganic salt is NaNO3、Na2CO3、NaHCO3One or more of sodium phosphate and potassium phosphate;
(2) the fatty acid ester released from the cells continuously has saponification reaction with alkali to generate fatty acid salt;
(3) filtering to remove algae cell debris;
(4) continuously adding inorganic salt solution into the system to separate out fatty acid salt, and filtering and collecting solid;
(5) adding inorganic acid into the fatty acid salt, acidifying to obtain free fatty acid, and collecting the product fatty acid.
2. The method of claim 1, wherein: the microalgae is derived from any green algae, diatom or red algae with the capacity of accumulating grease and fatty acid, and the collected microalgae is microalgae powder or microalgae mud.
3. The method of claim 2, wherein: the microalgae is chlorella or botryococcus.
4. The method of claim 1, wherein: the concentration of the inorganic salt in the step (1) is 0.01-0.03 mol/L.
5. The method of claim 1, wherein: the alkali solution in the step (1) is NaOH solution or KOH solution with the mass concentration of 10-30%.
6. The method of claim 1, 4 or 5, wherein: the mass-to-volume ratio (g: ml) of the microalgae collected in the step (1) to the inorganic salt solution and the alkali solution is 1: 5-1: 10, wherein the volume ratio of the inorganic salt solution to the alkali solution is 5: 1-1: 1.
7. The method of claim 1, wherein: the power of the microwave generator in the step (1) is 5-20 kW, the microwave frequency is 2450MHz or 915MHz, the radiation heating temperature is 50-70 ℃, and the heating time is 15-60 min.
8. The method of claim 1, wherein: the algae cell debris removed in step (3) can be membrane filtration or centrifugal filtration.
9. The method of claim 1, wherein: and (3) the inorganic salt in the step (4) is consistent with the inorganic salt added in the step (1) and is an unsaturated solution or a saturated solution with the concentration of 0.8-3 mol/L, and the volume-to-mass ratio (ml: g) of the added amount to the collected microalgae (the mass is calculated by dry weight) is 10: 1-5: 1.
10. The method of claim 1, wherein: and (4) adopting an inorganic salt saturated solution as the inorganic salt solution.
11. The method of claim 1, wherein: and (5) acidifying the inorganic acid in the step (5) to pH 1-4, wherein the inorganic acid is one or more of sulfuric acid, hydrochloric acid and nitric acid.
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| CN101586078A (en) * | 2009-06-18 | 2009-11-25 | 南京工业大学 | Scalization harvesting method and device for microalgae producing biological diesel oil |
| CN103045352A (en) * | 2011-10-17 | 2013-04-17 | 中国石油化工股份有限公司 | Extraction method of microalga grease |
| CN103396303A (en) * | 2013-07-25 | 2013-11-20 | 浙江大学 | Method for separating and purifying eicosapentaenoic acid and docosahexaenoic acid from micro-algal oil or fish oil |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101586078A (en) * | 2009-06-18 | 2009-11-25 | 南京工业大学 | Scalization harvesting method and device for microalgae producing biological diesel oil |
| CN103045352A (en) * | 2011-10-17 | 2013-04-17 | 中国石油化工股份有限公司 | Extraction method of microalga grease |
| CN103396303A (en) * | 2013-07-25 | 2013-11-20 | 浙江大学 | Method for separating and purifying eicosapentaenoic acid and docosahexaenoic acid from micro-algal oil or fish oil |
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