WO2019066146A1 - Procédé d'extraction de lipides à partir de microalgues - Google Patents

Procédé d'extraction de lipides à partir de microalgues Download PDF

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Publication number
WO2019066146A1
WO2019066146A1 PCT/KR2017/014750 KR2017014750W WO2019066146A1 WO 2019066146 A1 WO2019066146 A1 WO 2019066146A1 KR 2017014750 W KR2017014750 W KR 2017014750W WO 2019066146 A1 WO2019066146 A1 WO 2019066146A1
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microalgae
high shear
shear mixer
weight
organic solvent
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PCT/KR2017/014750
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English (en)
Korean (ko)
Inventor
장용근
곽민수
이봉수
문명훈
Original Assignee
재단법인 탄소순환형 차세대 바이오매스 생산전환 기술연구단
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Priority claimed from KR1020170171106A external-priority patent/KR102002575B1/ko
Application filed by 재단법인 탄소순환형 차세대 바이오매스 생산전환 기술연구단 filed Critical 재단법인 탄소순환형 차세대 바이오매스 생산전환 기술연구단
Publication of WO2019066146A1 publication Critical patent/WO2019066146A1/fr

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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
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats

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  • the present invention relates to a method for extracting lipids from microalgae. More particularly, the present invention relates to a method for extracting lipids from microalgae by administering a mixture containing 6 to 262 parts by weight of an organic solvent per 100 parts by weight of microalgae to a high shear mixer, To a method for extracting lipids from a sample.
  • Extraction processes include dry extraction and wet extraction. Wet processes are under study because the drying process required for dry extraction requires as much energy as it takes up more than 80% of the total biodiesel production process energy.
  • Scale-up is the easiest method for mechanical cell disruption using shear stress.
  • a high pressure homogenizer or bead milling has been mainly used for the lipid extraction method using the mechanical disruption method (Avinesh R. Byreddy et al., Bioresour Technol. 200: 464-9, 2016).
  • Both of these methods are capable of complete cell disruption and are easy to scale up.
  • a relatively large amount of solvent is required for extracting lipids, a large amount of energy is consumed, and when the cell disruption proceeds, There is a problem that the recovery is not performed smoothly.
  • the present inventors have made extensive efforts to develop a method for effectively extracting lipids by using a small amount of solvent. As a result, it has been found that when a high shear mixer is used, lipids can be efficiently extracted even with a small amount of solvent, It is confirmed that the emulsion is not generated in the extraction process, the lipid recovery is smooth, the lipid-removed microalgae and the organic solvent are separated by gravity, and the recovery of the organic solvent is easy. .
  • the present invention provides a method for producing a microalgae, comprising: (a) adding 6-262 parts by weight of an organic solvent to 100 parts by weight of microalgae; And (b) a method of extracting lipids from microalgae by administering a mixture of microalgae and an organic solvent to a high shear mixer.
  • 1 is a schematic diagram showing the principle of a high shear mixer in order.
  • FIG. 2 is a schematic diagram of an experiment for confirming the lipid extraction rate and energy consumption according to the stirring speed of the present invention and the ratio of microalgae to organic solvent.
  • FIG. 2 is a schematic diagram of an experiment for confirming the lipid extraction rate and energy consumption according to the stirring speed of the present invention and the ratio of microalgae to organic solvent.
  • FIG 3 is a graph showing the lipid extraction rate according to the stirring speed of the present invention.
  • FIG. 4 is a graph showing the lipid extraction rate of the microalgae (Orthioquitrium) of the present invention and the organic solvent (hexane) (volume-to-weight ratio) and stirring time.
  • FIG. 5 is a bar graph showing the lipid extraction rate according to the ratio (volume to volume ratio) of microalgae (Orthioquitrium) to organic solvent (hexane) after one hour treatment of the high shear mixer of the present invention.
  • FIG. 6 is a photograph showing that biomass and hexane are automatically separated over time after the lipid extraction of the present invention is completed.
  • FIG. 7 shows the result of lipid extraction from Orthioquitrium according to the extraction method.
  • FIG. 8 is a photograph of an emulsion produced according to the concentration of microalgae during lipid extraction according to the present invention.
  • lipids were not recovered in the extraction process, lipids were recovered smoothly, and lipid-removed microalgae and organic solvent were separated by gravity to facilitate recovery of organic solvent.
  • the present invention provides a method for producing a microalgae, comprising: (a) adding 6-262 parts by weight of an organic solvent to 100 parts by weight of microalgae; And (b) a method of extracting lipids from microalgae by administering a mixture of microalgae and an organic solvent to a high shear mixer.
  • the high shear mixer is a mixer made of a rotor-stator assembly
  • the shear rate of the high shear region is 20,000 to 100,000 s -1
  • the stirring speed is 2000 rpm to 10,000 rpm
  • Shear rate of 20000 s -1, or less than 2000 rpm of stirring speed is low shear.
  • shear rate (s -1 ) is the value obtained by dividing the rotor speed Vtip by the gap d between the rotor and the stator .
  • the high shear mixer can generate a high shear stress in the gap between the rotor and the stator and can operate at a higher rpm than a normal stirred tank, thus generating a strong turbulence flow. On the other hand, it shows weaker ability than high pressure homogenizer or bead milling, but it has the advantage that the energy consumption rate is reduced accordingly.
  • the high shear mixer is advantageous in that it is easy to disperse high viscosity materials with a strong mixing ability, is easy to scale up, and emulsion control is relatively easy (FIG. 1).
  • the blades of the high shear mixer of the present invention are of a cross shape, the diameter of the rotor is 31.44 mm, and the length of the gap between the rotor and the stator is 0.1 mm.
  • the relation between rpm and shear rate is as follows.
  • the stirring speeds of 1000 rpm, 3000 rpm, 5000 rpm, and 7000 rpm of the high shear mixer of the present invention can be represented by shear rates of 10960.7 s -1 , 32882 s -1 , 54803.3 s -1 and 76724.7 s -1 , respectively .
  • wet micro-algae can be efficiently dispersed into an organic solvent because a strong shear is generated.
  • lipid extraction rates of aurantiochytrium were compared using a high shear mixer method, a stirling method, and a sonication method.
  • the Sutering method showed no or no cell walls and a crushed Orantio kit It was confirmed that the lipid extraction rate was low even in rumen, and the sonication method and the high shear mixer method showed a lipid extraction rate of 100% (Fig. 7).
  • the microalgae and the organic solvent were used at a ratio of 1: 4 (volume ratio by weight).
  • the microalgae and the organic solvent were mixed at a ratio of 1: , It showed 100% lipid extraction rate.
  • the lipid extraction rate was 91%.
  • the sonication method showed that the biomass and hexane were not mixed efficiently even though the experiment was performed on a small scale and that the lipid extraction ratio was lower than that of the high shear mixer.
  • the method using the high shear mixer can extract 100% of the lipids even with a small amount of organic solvent, It can be seen that the high shear mixer is better.
  • the method of extracting lipids of the present invention can effectively extract lipids even with a small amount of solvent.
  • sonication is an authorized method that is usefully used to extract lipids from wet microalgae, and is able to effectively extract lipids through strong cell disruption capability.
  • the organic solvent may be selected from the group consisting of hexane, toluene, chloroform, acetone, heptane, dichloromethane, carbon tetrachloride and alcohol groups such as benzene, ethyl acetate, methanol, ethanol, isopropanol have.
  • the organic solvent is added in an amount of 6 to 262 parts by weight based on 100 parts by weight of the microalgae, and more specifically, the organic solvent is added in an amount of 32 to 196 parts by weight. do.
  • the density of hexane is 0.655 g / ml based on room temperature.
  • the change in lipid extraction rate with the amount of Aurantiochytrium versus hexane was observed.
  • High shear mixer did not show a decrease in the lipid extraction rate (Figs. 4 and 5), as compared to a 1: 1 ratio (volume to weight ratio) that decreased the lipid extraction rate. Therefore, it has been confirmed that the use of a high shear mixer can effectively extract lipids even when a small amount of organic solvent is used at the time of lipid extraction.
  • volume ratios by weight of Oranthiocritium and hexane in relation to the weight of 1: 4, 1: 3, 1: 2, 1: 1 and 1: 0.5 were 262 parts by weight, 196.5 parts by weight , 131 parts by weight, 65.5 parts by weight and 32.75 parts by weight.
  • volume ratio by weight of 1: 0.1 is 6.55 parts by weight based on 100 parts by weight of Orthioquitrium.
  • the present invention effectively extracts lipids and significantly reduces energy consumption even when using a small amount of an organic solvent.
  • the shear rate of the high shear mixer may be 20,000 to 100,000 s -1 , preferably 30000 to 80000 s -1 , and more preferably, Is 50000 to 60000 s < -1 >.
  • the lipid extraction rate according to the stirring speed of the high shear mixer was observed. As a result, it was confirmed that as the stirring speed increased, the lipid extraction rate increased and the lipid extraction rate reached 100% from 54803.3 s -1 (FIG. 3). In addition, at 76724.7 s -1 , the lipid extraction rate doubled, reaching a lipid extraction rate of 100% faster than 54803.3 s -1 , but a lot of heat was generated and the temperature rose to the boiling point of hexane causing a loss of hexane Table 1). Therefore, it was confirmed that the optimum stirring speed of the high shear mixer was 54803.3 s-1.
  • the step (a) may include the step of removing water from the microalgae.
  • the external water is removed through a centrifugation process using a vertical centrifuge.
  • the concentration of orantiocritium from which moisture is removed is 350 g / L.
  • micro-algae of 100 g / L and 200 g / L which do not remove water, were treated with a high shear mixer to confirm that the lipid extraction rate was lowered due to emulsion formation Respectively. Therefore, it can be seen that, in the lipid extraction using the high shear mixer, the emulsion is not formed and the lipid is extracted well by completely removing the moisture of the microalgae.
  • the external moisture of the microalgae is completely removed and the microalgae of high viscosity are dispersed in the organic solvent, and the biphasic extraction Respectively.
  • the microalgae may be wet microalgae.
  • the microalgae may be selected from the group consisting of Thraustochytrid, Chlamydomonas sp., Nannochloropsis sp. spores of the genus Chlorella sp., Chlorella sp., Scenedesmus sp. and Ettlia sp., can be characterized.
  • the Thraustochytrid system may include Aurantiochytrium sp., Schizochytrium sp., And Thraustochytrium sp. But is not limited thereto.
  • Example 1 Extraction of lipids using a high shear mixer
  • KRS101 was used.
  • the water content of the Orthioquitrium is 61.27 ⁇ 1.04%, and the total soil weight is 550 mg / g cell.
  • the KRS101 strain was cultured in a 5-ton incubator for 4 days, harvested and stored at 70 ° C.
  • technical grade hexane (95%) was used as the solvent for lipid extraction.
  • Chloroform, methanol, and sulfuric acid were used as the solvents for the lipid conversion, and the chloroform, methanol, and sulfuric acid were HPLC grade solvents.
  • the high shear mixer was a laboratory batch mixer L5M-A manufactured by Silverton, England. The experiment was performed using a glass beaker. The high shear mixer was placed 10 mm away from the left side of the beaker and 15 mm away from the bottom.
  • the amount of W l, W d, W b is the aluminum dish, the aluminum dish, and means the weight of the dried biomass and, V d and V t is the total solvent used in the amount of extraction experiments of solvents transferred to a dish lipids containing each . Based on the lipid content, the lipid extraction efficiency and the lipid extraction rate were calculated using the following equation.
  • the total lipid content refers to the total lipid content.
  • L 2 , L 1 , t 2 , and t 1 represent the later and first lipid extraction rates and time, respectively.
  • the moisture content and lipid content of the sample were measured as 64% of the weight of the wet microalgae and 55% of the weight of the dry microalgae on average.
  • Example 2 Change in lipid extraction rate according to shear rate of a high shear mixer
  • the change in lipid extraction rate with the shear rate of the high shear mixer was measured. 100 g of Aurantiochytrium and 400 ml of hexane were added to adjust the ratio of Aurantiochytrium and hexane to 1: 4 (by weight to volume ratio), and then the shear rate was increased to 10960.7 s - 1 , 32882 s -1 , 54803.3 s -1 and 76724.7 s -1 , and the lipid extraction rate was measured.
  • Table 1 shows the lipid extraction rate and the energy efficiency in the rest Portrait (shear rate) of 54803.3 and 76724.7 s -1 s -1.
  • Example 3 Change in lipid extraction rate according to the volume ratio of solvent to microalgae
  • Example 4 Energy efficiency of a high shear mixer
  • the power of the high shear mixer was measured using a Wattman HPM-100A meter. As a result, the lipid extraction rate was not significantly changed even when the amount of the solvent used was small, and only the stirring speed affects the lipid extraction rate.
  • the less the equivalent amount of hexane to decreased 3.3 times extracted Oran because a significant increase in the amount of thio kit Solarium (Aurantiochytrium) energy consumption is in 9.2MJ / kg to 2.8MJ / kg (Table 2).
  • Table 2 below shows the energy efficiency of the high shear mixer.
  • EROI energy consumption per unit dry biomass
  • EROI energy return on investment
  • Example 5 Identification of emulsion according to the concentration of microalgae in lipid extraction
  • the presence of emulsion was observed according to the concentration of microalgae in order to determine the optimum microalgae concentration.
  • 100 ml of hexane was added to 400 ml of Orthioquitrium at a concentration of 100 g / L, and Orantioquium and hexane were mixed at a ratio of 4: 1 (volume ratio by weight).
  • a high shear mixer After the treatment, the mixture was centrifuged at 5000 g for 5 minutes to confirm that hexane was separated without forming an emulsion.
  • the emulsion was better with increasing moisture content of microalgae, and the degree of emulsion was observed when treated for 10 minutes compared to 30 seconds of high shear mixer.
  • COMPARATIVE EXAMPLE 1 Comparison of lipid extraction ratio between high shear mixer method, stirling method and sonication method
  • the sonication was carried out with a 3 mm microtip at 30% amplitude for 1 hour using aurantiochytrium and hexane in a ratio of 1: 4 (weight to volume ratio), and the stuttering method was Orantiochitrium ( Aurantiochytrium ) and hexane at a ratio of 1: 4 (weight to volume ratio), the stirrer was treated at 500 rpm at 50 ° C for 1 hour. As a result, it was confirmed that the Sterling method had a low lipid extraction rate and the sonication method had a lipid extraction rate of 100% (FIG. 7).
  • the high shear mixer method was conducted for 1 hour under conditions of 54803.3 s -1 shear rate using a 1: 1 ratio (by volume to weight ratio) of aurantiochytrium and hexane. As a result, it was confirmed that the lipid extraction rate was 100% (Fig. 7).
  • the sonication method does not efficiently mix the biomass and hexane even though the experiment is conducted at a small scale, and the lipid extraction rate is lower than that of the high shear mixer.
  • the lipid can be extracted 100% It can be seen that the high shear mixer is better.
  • the method of extracting lipids from the microalgae of the present invention can effectively extract lipids even with a small amount of solvent and can significantly reduce the energy consumption. In the extraction process, no lipid is produced, It is easy to recover the organic solvent because the smooth, lipid-removed microalgae and the organic solvent are separated by gravity.

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Abstract

La présente invention concerne un procédé d'extraction de lipides à partir de microalgues et, plus spécifiquement, un procédé d'extraction de lipides à partir de microalgues par ajout, dans un mélangeur à cisaillement élevé, d'un mélange dans lequel 6 à 262 parts en masse d'un solvant organique sont ajoutées sur la base de 100 parts en masse de microalgues. La présente invention permet d'extraire efficacement des lipides, même lorsqu'une petite quantité de solvant est utilisée, et de réduire sensiblement la consommation d'énergie. De plus, les lipides sont récupérés sans à-coups étant donné qu'aucune émulsion n'est produite pendant le processus d'extraction, et étant donné que la biomasse, à partir de laquelle des lipides ont été éliminés, et le solvant organique sont séparés par gravité, la récupération d'un solvant organique est simple.
PCT/KR2017/014750 2017-09-26 2017-12-14 Procédé d'extraction de lipides à partir de microalgues WO2019066146A1 (fr)

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Application Number Priority Date Filing Date Title
KR20170124180 2017-09-26
KR10-2017-0124180 2017-09-26
KR1020170171106A KR102002575B1 (ko) 2017-09-26 2017-12-13 미세조류로부터 지질을 추출하는 방법
KR10-2017-0171106 2017-12-13

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112137110A (zh) * 2020-08-28 2020-12-29 盐城工学院 一种具有降血脂活性的沙蚕脂类的提取方法

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US20150251194A1 (en) * 2014-02-26 2015-09-10 James Madison Innovations, Inc. Method and apparatus for recovering non-hydrophilic components from algae-containing water
KR20150144397A (ko) * 2014-06-16 2015-12-28 한국과학기술원 양이온성 고분자와 전단 응력을 이용한 미세조류로부터 지질을 추출하는 방법 및 장치
KR20160053565A (ko) * 2014-11-05 2016-05-13 한국에너지기술연구원 양이온 계면활성제-자성나노입자 복합체를 이용한 미세조류의 수확 및 지질의 추출방법

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KR101317242B1 (ko) * 2013-07-30 2013-10-15 명지대학교 산학협력단 여과막을 이용한 미세조류로부터의 지질 추출 및 바이오 디젤 생산 방법
US20150251194A1 (en) * 2014-02-26 2015-09-10 James Madison Innovations, Inc. Method and apparatus for recovering non-hydrophilic components from algae-containing water
KR20150144397A (ko) * 2014-06-16 2015-12-28 한국과학기술원 양이온성 고분자와 전단 응력을 이용한 미세조류로부터 지질을 추출하는 방법 및 장치
KR20160053565A (ko) * 2014-11-05 2016-05-13 한국에너지기술연구원 양이온 계면활성제-자성나노입자 복합체를 이용한 미세조류의 수확 및 지질의 추출방법

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KWAK, M. ET AL.: "Extraction of Lipid from Wet Aurantiochytrium sp. KRS101 Using High Shear Mixer", KMB 2016 43RD ANNUAL MEETING & INTERNATIONAL SYMPOSIUM, June 2016 (2016-06-01) *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112137110A (zh) * 2020-08-28 2020-12-29 盐城工学院 一种具有降血脂活性的沙蚕脂类的提取方法

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