KR20000000842A - Extraction method of hydrocarbon from microalgae - Google Patents

Extraction method of hydrocarbon from microalgae Download PDF

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KR20000000842A
KR20000000842A KR1019980020707A KR19980020707A KR20000000842A KR 20000000842 A KR20000000842 A KR 20000000842A KR 1019980020707 A KR1019980020707 A KR 1019980020707A KR 19980020707 A KR19980020707 A KR 19980020707A KR 20000000842 A KR20000000842 A KR 20000000842A
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microalgae
hydrocarbon
methanol
mixed solvent
chloroform
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KR1019980020707A
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KR100320786B1 (en
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오희목
이석준
윤병대
김희식
안극현
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박호군
한국과학기술연구원
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D39/00—Filtering material for liquid or gaseous fluids
    • B01D39/14—Other self-supporting filtering material ; Other filtering material
    • B01D39/16—Other self-supporting filtering material ; Other filtering material of organic material, e.g. synthetic fibres
    • B01D39/1607—Other self-supporting filtering material ; Other filtering material of organic material, e.g. synthetic fibres the material being fibrous
    • B01D39/1623—Other self-supporting filtering material ; Other filtering material of organic material, e.g. synthetic fibres the material being fibrous of synthetic origin
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D27/00—Cartridge filters of the throw-away type
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D39/00—Filtering material for liquid or gaseous fluids
    • B01D39/14—Other self-supporting filtering material ; Other filtering material
    • B01D39/16—Other self-supporting filtering material ; Other filtering material of organic material, e.g. synthetic fibres
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2239/00—Aspects relating to filtering material for liquid or gaseous fluids
    • B01D2239/06—Filter cloth, e.g. knitted, woven non-woven; self-supported material
    • B01D2239/0604—Arrangement of the fibres in the filtering material
    • B01D2239/0618—Non-woven
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2239/00—Aspects relating to filtering material for liquid or gaseous fluids
    • B01D2239/06—Filter cloth, e.g. knitted, woven non-woven; self-supported material
    • B01D2239/0604—Arrangement of the fibres in the filtering material
    • B01D2239/0622—Melt-blown
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2239/00—Aspects relating to filtering material for liquid or gaseous fluids
    • B01D2239/06—Filter cloth, e.g. knitted, woven non-woven; self-supported material
    • B01D2239/0604—Arrangement of the fibres in the filtering material
    • B01D2239/0627—Spun-bonded

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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

PURPOSE: An extraction method of massive hydrocarbon from Botryoccoccus barunii in a short time is provided which is time and cost effective. CONSTITUTION: Hydrocarbon is extracted by: culturing and recovering microalgae, Botryoccoccus barunii UEX 572; dispersing Botryoccoccus barunii UEX 572 into a solvent, which is a mixture of chloroform and methanol in a weight ratio of 2:1 and breaking the microalgae by using glass beads; extracting hydrocarbon and measuring its content.

Description

미세조류로부터 탄화수소의 추출방법Extraction of hydrocarbons from microalgae

본 발명은 미세조류로부터 탄화수소를 추출하는 방법에 관한 것이다. 더욱 상세하게는, 본 발명은 미세조류 보트리오코커스 브라우니(Botryococcus braunii) UTEX 572를 이용하여 클로로포름과 메탄올의 혼합용매에 분산시키고, 유리비드로 파쇄하여 탄화수소를 추출하는 방법에 관한 것이다.The present invention relates to a method for extracting hydrocarbons from microalgae. More specifically, the present invention relates to a method of dispersing a hydrocarbon by dispersing in a mixed solvent of chloroform and methanol using a microalgae Botryococcus braunii UTEX 572, and crushing with glass beads.

보트리오코커스속 미세조류는 지름 0.5㎜ 크기의 군체를 이루어 성장하는 녹조류로 온대와 열대지방에 걸쳐 광범위하게 분포한다. 보트리오코커스속 미세조류는 올레인산(oleic acid)으로부터 탄소수 34의 불포화 테르펜계 탄화수소(hydrocarbon)인 보트리오코커센(botryococcene)과 이소보트리오코커센(isobotryococcene)을 축적하는데, 생장기의 녹색세포는 탄소수 27, 29, 31의 디엔(diene)류를 생산한다. 이들 탄화수소류들의 화학적 조성은 자동차 연료인 디젤유의 조성과 비슷하여, 수송연료로 사용할 석유자원을 대체하는 에너지원으로 많은 연구가 진행되고 있다(Hillen L.W. et al., Biotech. Bioeng., 24, 193, 1982; Knight B.A. et al., Phytochem., 9, 1317, 1970). 그러나, 미세조류는 균체내부에 지방함량이 높아 담수나 염분이 함유된 물의 표면에서 부유하는 형태로 생장하고, 또한 미세조류의 종류에 따라 물리·화학적 조성이 다르기 때문에 산업적으로 활용하기 위한 공정개발에 어려움이 따른다(Buelna G. et al., Biological Wastes, 31, 211, 1990). 또한 미세조류로부터 탄화수소를 추출함에 있어서 종래의 방법으로는 미세조류를 동결건조하고, 용매로 탄화수소를 추출하는 동결건조방법이 있으나 비용과 시간이 많이 소요되는 비경제적인 추출방법이다(Ahlgren, G. and Uppsala, L.M., Arch Hydrobiol., 121, 295-306, 1991). 따라서, 본 발명자들은 이러한 문제점을 극복하기 위하여 미세조류, 특히 보트리오코커스 브라우니(Botryococcus braunii) UTEX 572의 배양액을 pH 11로 조정하여 미세조류의 균체를 침전시켜 탄수화물을 회수하는 방법을 개발한 바 있으나(한국특허출원 제 97-61307호), 본 발명은 이보다 더욱 추출이 용이한 방법을 제공하고자 하였다. 따라서, 본 발명의 목적은 상기와 같은 문제점과 편의성을 감안하여 안출한 것으로 클로로포름과 메탄올의 혼합용매에 미세조류 보트리오코커스 브라우니(Botryococcus braunii) UTEX 572를 분산시킨 후 유리비드를 이용하여 균체의 두꺼운 세포벽을 용이하게 파괴하므로써 적은 시간과 경제적인 비용으로 탄화수소를 대량 추출하여 제공함에 있다.The microalgae of Boturiococcus are green algae that grow in colonies with a diameter of 0.5 mm and are widely distributed in temperate and tropical regions. The microalgae of Boturiococcus accumulate botryococcene and isobotryococcene, which are unsaturated terpene hydrocarbons of 34 carbon atoms, from oleic acid. It produces 27, 29 and 31 dienes. The chemical composition of these hydrocarbons is similar to that of diesel oil, which is a vehicle fuel, and much research is being conducted as an energy source to replace petroleum resources for transportation fuels (Hillen LW et al., Biotech. Bioeng., 24, 193). , 1982; Knight BA et al., Phytochem., 9, 1317, 1970). However, microalgae grow in the form of floating on the surface of fresh water or salt-containing water due to the high fat content in the cells, and the physical and chemical composition varies depending on the type of microalgae. Difficulties follow (Buelna G. et al., Biological Wastes, 31, 211, 1990). In addition, conventional methods for extracting hydrocarbons from microalgae are lyophilization methods for lyophilizing microalgae and extracting hydrocarbons with solvents, but are an uneconomical and costly extraction method (Ahlgren, G. and Uppsala, LM, Arch Hydrobiol., 121, 295-306, 1991). Therefore, the present inventors have developed a method for recovering carbohydrates by adjusting the culture of microalgae, especially Botryococcus braunii UTEX 572 to pH 11 to precipitate the microalgae cells. (Korean Patent Application No. 97-61307), the present invention was intended to provide a method that is easier to extract than this. Accordingly, an object of the present invention was devised in view of the above problems and conveniences, and after dispersing the microalgae Botryococcus braunii UTEX 572 in a mixed solvent of chloroform and methanol, the cells were thickened using glass beads. By easily breaking the cell wall, a large amount of hydrocarbons are extracted and provided in a small amount of time and economic cost.

본 발명의 상기 목적은 클로로포름과 메탄올이 일정비율로 혼합된 혼합용매에 미세조류 보트리오코커스 브라우니(Botryococcus braunii) UTEX 572를 분산시킨 후, 유리비드로 미세조류를 파쇄하므로써 달성하였다.The object of the present invention was achieved by dispersing the microalgae Botryococcus braunii UTEX 572 in a mixed solvent in which chloroform and methanol were mixed at a constant ratio, and then crushing the microalgae with glass beads.

이하, 본 발명의 구성 및 작용을 상세히 설명한다.Hereinafter, the configuration and operation of the present invention will be described in detail.

본 발명은 미세조류의 배양 및 회수단계; 상기 회수된 미세조류에 클로로포름과 메탄올의 혼합용매를 사용하여 분산시킨 후 유리비드를 이용하여 세포벽을 파괴하고 탄화수소를 추출한 후, 추출된 탄화수소 함량을 측정하는 단계; 및/또는 상기 회수된 미세조류에 클로로포름과 메탄올의 혼합용매와 또다른 종류의 다양한 혼합용매를 사용하여 분산시킨 후 탄화수소 함량을 측정하여 탄화수소 추출 효율을 비교 분석하는 단계로 이루어진다.The present invention is culture and recovery step of microalgae; Dispersing the recovered microalgae using a mixed solvent of chloroform and methanol, breaking the cell wall using glass beads, extracting hydrocarbons, and then measuring the extracted hydrocarbon content; And / or dispersing the mixed microalgae using a mixed solvent of chloroform and methanol and another mixed solvent of another kind, and then measuring the hydrocarbon content to compare and analyze the hydrocarbon extraction efficiency.

이하, 본 발명의 구체적인 구성 및 작용을 실시예 및 실험예를 통하여 상세히 설명하지만 본 발명의 권리범위가 이들 실시예에 의해 제한되는 것은 아니다.Hereinafter, the specific configuration and operation of the present invention will be described in detail with reference to examples and experimental examples, but the scope of the present invention is not limited to these examples.

실시예 1. 미세조류의 배양 및 회수Example 1 Cultivation and Recovery of Microalgae

텍사스 대학에서 분양받은 보트리오코커스 브라우니(Botryococcus braunii) UTEX 572를 Chu 13 배지(KNO3371㎎, K2HPO480㎎, MgSO4·7H2O 200㎎, CaCl2·2H2O 107㎎, ferric citrate 20㎎, citric acid 100㎎, H3BO45.72㎎, MnCl2·4H2O 3.62㎎, ZnSO4·7H2O 0.44㎎, CuSO4·5H2O 0.16㎎, CoCl20.02㎎, Na2MoO40.084㎎/L, pH 7.5)에서 25℃, 100rpm, 100μE/㎡/s의 광도하에 호기적인 상태로 5주간 배양하였다. 이어서, 수산화나트륨 용액을 가하여 pH 11로 조절하여 침전시킨 다음, 원심분리하여 미세조류를 회수하였다. 회수한 미세조류는 분석에 사용할 때까지 냉장고에서 보관하였다.Botryococcus braunii UTEX 572, obtained from the University of Texas, was prepared using Chu 13 medium (KNO 3 371 mg, K 2 HPO 4 80 mg, MgSO 4 7H 2 O 200 mg, CaCl 2 2H 2 O 107 mg, ferric citrate 20 mg, citric acid 100 mg, H 3 BO 4 5.72 mg, MnCl 2 · 4H 2 O 3.62 mg, ZnSO 4 · 7H 2 O 0.44 mg, CuSO 4 · 5H 2 O 0.16 mg, CoCl 2 0.02 mg, Na 2 MoO 4 0.084 mg / L, pH 7.5) for 5 weeks under aerobic conditions at 25 ° C., 100 rpm, and 100 μE / m 2 / s of luminous intensity. Subsequently, sodium hydroxide solution was added to adjust pH to 11, followed by centrifugation to recover microalgae. The recovered algae was stored in the refrigerator until it was used for analysis.

실시예 2. 탄화수소 함량 측정Example 2. Hydrocarbon Content Determination

상기 실시예 1에서 회수한 미세조류를 클로로포름과 메탄올을 2:1중량부로 혼합한 혼합용매에 분산시키고, 비드파쇄기(Biospec사 제품, 모델 3110BX, 유리비드 입자 1㎜)를 사용하여 30초간 미세조류를 파쇄하고, 소금을 5% 되도록 첨가하여 층을 분리한 다음, 탄화수소 함량 측정법(Bligh, E. G. and Dyer, W. J., Can. J. Biochem. Physiol., 37:911 - 917, 1959)에 따라 상층부를 회수하고, 용매를 휘발시킨 다음 잔존된 탄화수소 함량을 측정하였다. 실험결과, 탄화수소 함량은 2.27±0.16g/L 였으며, 초기에 투입된 미세조류의 건조량은 7.92g/L 로, 5회 반복실험에서 탄화수소 함량은 미세조류 건조중량의 28.67±1.98% 범위였다.The microalgae recovered in Example 1 was dispersed in a mixed solvent in which chloroform and methanol were mixed at 2: 1 parts by weight, and the microalgae were used for 30 seconds using a bead crusher (Biospec, model 3110BX, glass bead particles 1 mm). Crushed, the salt was added to 5% to separate the layers, and the upper layer was prepared according to the hydrocarbon content measurement method (Bligh, EG and Dyer, WJ, Can. J. Biochem. Physiol., 37: 911-917, 1959). After recovery, the solvent was volatilized and the remaining hydrocarbon content was measured. As a result, the hydrocarbon content was 2.27 ± 0.16g / L, and the amount of dried microalgae initially was 7.92g / L. In five replicates, the hydrocarbon content was 28.67 ± 1.98% of the dry weight of the microalgae.

실시예 3. 탄화수소 함량 비교Example 3. Comparison of Hydrocarbon Content

실험예 1 : 동결건조기를 이용한 탄화수소 추출량 측정Experimental Example 1 Measurement of Hydrocarbon Extraction Using a Freeze Dryer

상기 실시예 1에서 얻은 미세조류를 동결건조기(삼원사 제품, 모델 SFDSM06)로 동결 건조하여 미세조류를 파쇄한 다음, 상기 실시예 2의 혼합용매를 사용하여 탄화수소 함량을 측정하였다. 실험결과, 탄화수소 함량은 미세조류 건조중량의 19.56±0.58% 범위였다.The microalgae obtained in Example 1 was lyophilized with a lyophilizer (manufactured by SAMWON, model SFDSM06) to crush the microalgae, and the hydrocarbon content was measured using the mixed solvent of Example 2. As a result, the hydrocarbon content was in the range of 19.56 ± 0.58% of the microalgae dry weight.

실험예 2 : 초음파기를 이용한 탄화수소 추출량 측정Experimental Example 2 Measurement of Hydrocarbon Extraction Rate Using an Ultrasonic Machine

상기 실시예 1에서 얻은 미세조류를 상기 실시예 2의 혼합용매에 분산시키고, 초음파기(Sonics & Materials Inc. 제품, 모델 Vibra Cell), 호모게나이저(Nihon Seiki사 제품, 모델 BM-3), 프렌치 프레서(SLM Instruments사 제품, 모델 SLM Aminco)를 각각 사용하여 미세조류를 파쇄한 후 탄화수소 함량을 측정하였다. 실험결과, 탄화수소 함량은 미세조류의 건조중량당 초음파 파쇄의 경우 16.52±0.87%, 호모게나이저를 이용하여 파쇄한 경우 14.95±1.85%, 프렌치 프레서를 이용하여 파쇄한 경우 22.49±2.69% 범위였다.The microalgae obtained in Example 1 were dispersed in the mixed solvent of Example 2, sonicator (Sonics & Materials Inc., model Vibra Cell), homogenizer (Nihon Seiki, model BM-3), French The hydrocarbon content was measured after crushing the microalgae using a presser (SLM Instruments, model SLM Aminco). As a result, the hydrocarbon content ranged from 16.52 ± 0.87% for ultrasonic crushing per microalgae, 14.95 ± 1.85% for crushing using homogenizer, and 22.49 ± 2.69% for crushing using French presser.

실험예 3 : 혼합용매를 사용한 탄화수소 추출량 측정Experimental Example 3 Measurement of Hydrocarbon Extraction Using Mixed Solvent

상기 실시예 2와 동일한 혼합용매를 사용하여 미세조류를 파쇄하지 않고 직접 혼합하여 탄화수소를 추출하였다. 실험결과, 탄화수소의 함량은 미세조류 건조중량의 14.73±0.90% 였다.Hydrocarbons were extracted by directly mixing the microalgae without crushing using the same mixed solvent as in Example 2. As a result, the hydrocarbon content was 14.73 ± 0.90% of the dry weight of the microalgae.

실험예 4 : 추출용매의 종류에 따른 탄화수소 추출량 측정Experimental Example 4 Measurement of Hydrocarbon Extraction According to the Type of Extraction Solvent

미세조류로부터 추출되는 탄화수소의 함량은 추출용매의 종류에 따라서도 달라지므로 보다 효율적인 추출용매를 선발하기 위하여 여러 가지 추출용매를 사용하여 탄화수소의 함량을 비교하였다. 즉, 상기 실시예 2의 추출용매를 헥산/이소프로판올 3:2 중량부, 디클로로에탄/메탄올 1:1 중량부, 디클로로메탄/에탄올 1:1 중량부, 아세톤/디클로로메탄 1:1 중량부의 비율로 각각 혼합된 혼합용매로 대체하여 비드파쇄기로 파쇄한 후 각각의 추출용매를 사용하여 미세조류로부터 추출된 탄화수소 함량을 측정하였다. 실험결과, 탄화수소 함량은 헥산/이소프로판올 3:2의 경우 20.23±0.59%, 디클로로에탄/메탄올 1:1의 경우 18.64±1.41%, 디클로로메탄/에탄올 1:1의 경우 18.83±0.93, 아세톤/디클로로메탄 1:1의 경우 18.35±1.06% 였다.The content of hydrocarbons extracted from the microalgae also varies depending on the type of extraction solvent, and the content of hydrocarbons was compared using various extraction solvents to select more efficient extraction solvents. That is, the extraction solvent of Example 2 in the ratio of 3: 2 parts by weight of hexane / isopropanol, 1: 1 parts by weight of dichloroethane / methanol, 1: 1 parts by weight of dichloromethane / ethanol, and 1: 1 parts by weight of acetone / dichloromethane. The contents of hydrocarbons extracted from the microalgae were measured using each extractant after crushing with a bead crusher in place of each mixed solvent. Experimental results show that the hydrocarbon content is 20.23 ± 0.59% for hexane / isopropanol 3: 2, 18.64 ± 1.41% for dichloroethane / methanol 1: 1, 18.83 ± 0.93 for dichloromethane / ethanol 1: 1, acetone / dichloromethane In the case of 1: 1, it was 18.35 ± 1.06%.

따라서, 미세조류로부터 탄화수소를 추출하는데 있어서, 미세조류를 파쇄할 때 유리비드를 사용하고, 클로로포름과 메탄올의 혼합용매를 사용하여 추출하는 것이 가장 효율적 방법임을 알 수 있었다.Therefore, in extracting hydrocarbons from the microalgae, it was found that the extraction of the algae using glass beads and the mixed solvent of chloroform and methanol was the most efficient method.

본 발명은 상기의 실시예를 통하여 알 수 있듯이, 미세조류 보트리오코커스 브라우니(Botryococcus braunii) UTEX 572에서 클로로포름과 메탄올이 2:1의 중량부로 혼합된 혼합용매와 유리비드를 이용하여 적은비용으로 다량의 탄화수소를 용이하게 추출하는 효과가 있으므로 생물산업상 유용한 발명인 것이다. 또한 본 발명은 보트리오코커스 브라우니(Botryococcus braunii) UTEX 572에서 추출한 다량의 탄화수소를 대체 에너지 자원으로 이용할 수 있는 효과가 기대되므로 에너지 산업상 매우 유용한 발명인 것이다.As can be seen from the above embodiment, the microalgae Botryococcus braunii UTEX 572 in a large amount at a low cost by using a mixed solvent and glass beads in which chloroform and methanol are mixed in a 2: 1 part by weight Since it has the effect of easily extracting the hydrocarbon of the invention is useful in the biological industry. In addition, the present invention is a very useful invention in the energy industry because it is expected that the effect of using a large amount of hydrocarbon extracted from Botryococcus braunii UTEX 572 as an alternative energy source.

Claims (2)

미세조류를 클로로포름과 메탄올이 2:1 중량비로 혼합된 혼합용매에 분산시키고 유리비드를 이용하여 상기 미세조류를 파쇄함을 특징으로 하는 미세조류로부터 탄화수소의 추출방법.A method of extracting hydrocarbons from microalgae, wherein the microalgae are dispersed in a mixed solvent in which chloroform and methanol are mixed in a 2: 1 weight ratio, and the microalgae are crushed using glass beads. 제1항에 있어서, 상기 미세조류가 보트리오코커스속임을 특징으로 하는 방법.The method of claim 1 wherein the microalgae are of the genus Boliococcus.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100672573B1 (en) * 2004-10-14 2007-01-24 엘지전자 주식회사 Control assembly in electric range
US9102972B2 (en) 2012-03-05 2015-08-11 Korea Research Institute Of Bioscience And Biotechnology Microorganism Rhizobium sp. KB10 having properties of promoting growth of Botryococcus braunii and increasing fatty acid content
WO2015152620A1 (en) * 2014-04-01 2015-10-08 인하대학교 산학협력단 High-performance empirical analysis of metabolic pathways in microalgae using cell-free system

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JPH05301097A (en) * 1992-04-24 1993-11-16 Agency Of Ind Science & Technol Method for purification of treated sewage using microalgae and for simultaneous production of hydrocarbon
JPH09803A (en) * 1995-06-19 1997-01-07 Chikyu Kankyo Sangyo Gijutsu Kenkyu Kiko Method for extracting hydrocarbons from minute algae belonging to botryococcus
JPH09234055A (en) * 1996-03-01 1997-09-09 Chikyu Kankyo Sangyo Gijutsu Kenkyu Kiko New fine alga belonging to genus botryococcus and its culture

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100672573B1 (en) * 2004-10-14 2007-01-24 엘지전자 주식회사 Control assembly in electric range
US9102972B2 (en) 2012-03-05 2015-08-11 Korea Research Institute Of Bioscience And Biotechnology Microorganism Rhizobium sp. KB10 having properties of promoting growth of Botryococcus braunii and increasing fatty acid content
WO2015152620A1 (en) * 2014-04-01 2015-10-08 인하대학교 산학협력단 High-performance empirical analysis of metabolic pathways in microalgae using cell-free system

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