CN103882069A - Preparation method of long chain fatty acid ester - Google Patents
Preparation method of long chain fatty acid ester Download PDFInfo
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Abstract
The invention discloses a preparation method of long chain fatty acid ester, and belongs to the downstream technical field of bio-engineering. In the presence of water, oil-producing microorganisms and short chain alcohol are taken as the raw materials, and long chain fatty acid ester is obtained through the self-catalytic conversion of microorganisms. The fermenting mash of oil-producing microorganisms and fermenting mash containing short chain alcohol can be directly taken as the raw materials, the microorganisms have a strong water tolerance ability during the conversion process, no additional catalysis is required, the operation conditions are mild, the bacterial strains do not need to be subjected to complicated genetic engineering modification, and thus the preparation method has the advantages of simple process, easy amplification, low requirements on equipment, low energy consumption, low cost, and environment-friendliness. The invention provides an economic and feasible novel technology for massive production of long chain fatty acid ester.
Description
Technical field
The present invention relates to a kind of preparation method of long chain fatty acid ester, concretely, is under the environment that has water to exist, take the thalline of oleaginous microorganism and short chain alcohol as raw material, transform through thalline autocatalysis, generate long chain fatty acid ester, belong to biotechnology downstream technical field.
Background technology
By animal and plant grease or longer chain fatty acid and short chain alcohol (as methyl alcohol and ethanol) reaction, obtain long chain fatty acid ester, the current biofuel that is often referred to as.Biofuel energy density is high, lubricity good, storing and transporting security, anti-knocking property are good, sufficient combustion, renewable, environmental friendliness, can directly apply to diesel engine system, is one of desirable biofuel kind.Vegetable oil resource is very limited, and its production is subject to the impact of place, season and climate change, and expand oil crops plantation may exist with people strive grain, with grain strive etc. problem.Therefore, there is lack of raw materials becomes one of bottleneck of restriction Biodiesel scale.Part microorganism can be at intracellular accumulation grease, and can reach the more than 20% of its dry cell weight, this quasi-microorganism is called as oleaginous microorganism (Ratledge C, Wynn JP.The biochemistry and molecular biology of lipidaccumulation in oleaginous microorganisms.Advances in Applied Microbiology, 2002,51,1).Some kinds in fungi, bacterium and micro-algae have the ability of excess accumulation grease, meet the feature of oleaginous microorganism.The grease that oleaginous microorganism is synthetic, is called microbial oil, conventionally has the lipid acid composition close with animal and plant grease.Compared with animal and plant grease technology, the advantage such as that microbial oil technology has is with short production cycle, can produce continuously, production potential are large.Because oleaginous microorganism can be converted into grease by the organism such as carbohydrate that derives from biomass resource, so microbial oil raw material resources are abundant and cheap.Therefore, microbial oil be solve that Biodiesel raw material problem is the most effective, one of continuable approach (Zhao Zongbao. Chinese biological engineering magazine, 2005,25,8).
Produce long chain fatty acid ester based on microbial oil technology, can be divided into two primary processes, i.e. the production of microbial oil and the production of long chain fatty acid ester.Previous process generally includes oleaginous microorganism cultivation and extracts and separate with microbial oil, and a rear process is to complete by catalyst or in super (Asia) critical fluids.The catalyzer that long chain fatty acid ester is prepared in the conversion of catalysis grease has acid, alkali, lipase and ionic liquid etc.Acid catalyst comprises free acid, solid acid etc., and alkaline catalysts comprises mineral alkali, solid alkali etc., and lipase-catalyzed dose comprises resolvase, immobilized enzyme and immobilized cell, and ionic-liquid catalyst comprises dissociated ion liquid and immobilization ionic liquid.If above-mentioned catalysis transesterification or esterification will obtain more than 60% high-level efficiency, need to be in moisture content lower than 30%(w/w) condition under carry out (AtadashiIM, Aroua MK, AzizARA, Sulaiman NMN.Renewable and Sustainable Energy Reviews, 2012,16,3456).Prepare long chain fatty acid ester except using catalyst, can also in super (Asia) critical methanol, water or other fluid, carry out.This process does not need to add catalyzer, higher to the tolerance of moisture.For example, at 350 ° of C, under 43MPa, the water-content of supercritical methanol-aqueous systems exceedes 30%(w/w) time, the yield of fatty acid methyl ester still can reach more than 90% (Kusdiana D, Saka S.Bioresource Technology, 2004,91,289).Oleaginous microorganism is cultivated the fermentation liquid obtaining, and moisture content is conventionally more than 80%, so will carry out processed before catalyzed reaction.Research shows, the energy that moisture removal consumes accounts for long chain fatty acid ester and produces the more than 80% of total energy consumption (Teixeira RE.Green Chemistry, 2012,14,419).In super (Asia) critical fluids, produce long chain fatty acid ester, although allow higher moisture content, need under High Temperature High Pressure, carry out, high to equipment requirements, energy consumption is large, and cost is high, and other side reactions also may occur for other compositions in fermentation liquid and thalline.Therefore, need to set up the novel method of preparing long chain fatty acid ester of mating with microbial oil technology, the directly raw material of trans-utilization high moisture content under mild conditions, significantly to improve the Technical Economy of process.
Researchist started to attempt under normal temperature and pressure conditions in recent years, utilized microorganism directly synthetic long chain fatty acid ester in high moisture environments, and this kind of method need to be carried out genetic engineering modified to microorganism conventionally.For example; heterogenous expression wax ester synthetic enzyme/acyl CoA in yeast saccharomyces cerevisiae: diacylglycerol acyltransferase gene (atfA) is also added lipid acid during the fermentation; can obtain long-chain fat acetoacetic ester (Kalscheuer R; LuftmannH; Steinbuchel A.Applied and Environmental Microbiology; 2004,70,7119).At the gene of expression in escherichia coli atfA gene and coding pyruvic carboxylase and ethanol dehydrogenase, by adding lipid acid, in intestinal bacteria, long-chain fat acetoacetic ester content reaches the 25.4%(Elbahloul Y.Steinbuechel A.Applied and Environmental Microbiology of dry cell weight, 2010,76,4560).On the basis of expression in escherichia coli atfA gene and ethanol synthase gene, by transformation fatty acid metabolism path, can realize production (the Steen EJ to long-chain fat acetoacetic ester by monose, Kang, YS, Keasling JD, et al.Nature, 2010,463,559), after this, by introducing dynamic pickup regulator control system, long-chain fat acetoacetic ester output reaches 1.5g/L(Zhang FZ, Carothers JM, Keasling JD.Nature Biotechnology, 2012,30,354).Utilize directly synthetic long-chain fat acetoacetic ester of microorganism, can carry out at normal temperatures and pressures, but bacterial strain need to be genetic engineering modified, process adjustment complexity.For the long chain fatty acid ester of other short chain alcohol (as methyl alcohol, propyl alcohol etc.), owing to being subject to the restriction of microbial metabolism approach, not yet building and obtain efficient gene engineering strain at present.
Summary of the invention
The invention provides under a kind of environment there being water to exist, take the thalline of oleaginous microorganism and short chain alcohol as raw material, transform through thalline autocatalysis, generate the method for long chain fatty acid ester.The present invention can be directly take oleaginous microorganism fermentation liquid and the fermentation liquid that contains short chain alcohol as raw material, conversion process is strong to moisture tolerance, do not need to add extra catalyzer, operational condition gentleness, does not need bacterial strain to carry out the genetic engineering modified of complexity, process is simple, easily amplify, low for equipment requirements, energy consumption is low, cost is low, free from environmental pollution.
The present invention is achieved by following technical proposals:
1) as follows a kind of or the combination of their necessity, preparation feedback system:
A) in the fermentation liquid that contains oleaginous microorganism thalline, add short chain alcohol, reach 0.1%～90% to the concentration expressed in percentage by volume of short chain alcohol; Or
B) thalline of water, oleaginous microorganism and short chain alcohol are mixed, reach 0.1%～90% to the concentration expressed in percentage by volume of short chain alcohol, the mass percentage concentration of water is 10%～99%; Or
C) fermentation liquid that contains oleaginous microorganism thalline and the fermented liquid that contains short chain alcohol are mixed by a certain percentage, reach 0.1%～15% to the concentration expressed in percentage by volume of short chain alcohol; Or
D) thalline of water, oleaginous microorganism and the fermented liquid that contains short chain alcohol are mixed by a certain percentage, reach 0.1%～15% to the concentration expressed in percentage by volume of short chain alcohol.
2) potential of hydrogen of the reaction system of being prepared by step 1) is adjusted to pH2～10, under 20 ° of C～60 ° C of temperature, processes 5 minutes～168 hours;
3) to step 2) add organic solvent to extract in the system of gained, remove organic solvent and obtain long chain fatty acid ester.
The fermentation liquid that contains oleaginous microorganism thalline of the present invention is the suspension liquid that contains thalline obtaining after oleaginous microorganism fluid suspension culture; The thalline of oleaginous microorganism, for the wet thallus separated from oleaginous microorganism culture system or through dehydrating the dry mycelium of processing.
Short chain alcohol of the present invention is one or more necessity combination in methyl alcohol, ethanol, propyl alcohol, butanols, amylalcohol.
The reagent of adjustment reaction system potential of hydrogen of the present invention is conventional sour example hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid etc., or conventional alkali is as sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate, salt of wormwood, ammoniacal liquor etc.
Organic solvent of the present invention is the liquid state organics that polarity is lower, as sherwood oil, normal hexane, ethyl acetate, methylene dichloride, chloroform, No. six solvent for extraction, No. four solvents, industrial hexane etc. or their necessity combination.
The present invention removes the method that organic solvent obtains long chain fatty acid ester mutually from organic solvent be air distillation or vacuum-evaporation etc.
In order to improve the technique effect of step 3) of the present invention, before enforcement solvent extraction or in leaching process, also can be aided with the physics and chemistry processing such as heating, microwave radiation, supersound process, enzyme processing, acid treatment, high-pressure homogeneous processing, ball-milling processing, granulated glass sphere fragmentation, multigelation, to destroy oleaginous microorganism cell, improve the long chain fatty acid ester rate of recovery.
Oleaginous microorganism thalline of the present invention is eukaryotic microorganisms thalline or the prokaryotic micro-organisms thalline that fat content exceedes its dry weight 20%.Exceed dry cell weight 20%(w/w for the preparation of the oleaginous microorganism of oleaginous microorganism thalline for thalline fat content after fermentation culture) eukaryotic microorganisms and prokaryotic micro-organisms in one or more necessity combination, they include but not limited to, produce oil fungi, as red winter spore yeast (Rhodosporidium sp.), Cryptococcus (Cryptococcus sp.), rhodotorula (Rhodotorula sp.), saccharomyces oleaginosus (Lipomyces sp.), trichosporon (Trichosporon sp.), sub-sieve solution fat yeast (Yarrowialipolytica), strong mould (Geotrichum robustum) doughtily, Mortierella isabellina (Mortierella isabellina), volume branch Mucor (Mucor circinelloides), Cunninghamella sp (Cunninghamella) and Christian Breton endomycopsi.sp (Endomycopsis burtonii), oil-producing microalgae, as Botryococcus braunii (Botryococcus braunii), Crypthecodinium cohnii (Crypthecodinium cohnii), chlorella (Chlorellaprotothecoides), Nannochloropsis oceanica (Nannochloropsis sp.) and schizochytrium limacinum (Schizochytrium limacinum), produce oil bacterium, as coryneform bacteria (Corynebacterium), Nocardia bacteria (Nocarda), mycobacterium (Mycobacterium) etc.
The present invention uses the actual fat content of thalline material to measure the method for reference literature (Li YH, Zhao ZB, BaiFW.Enzyme Microbial Technology, 2007,41,312).The method of the evaluation of long chain fatty acid ester and content detection reference literature (Gu HQ, Jiang YJ, Zhou LY, Gao J.Energy & EnvironmentalScience, 2011,4,1337) is carried out.
Embodiment
Following examples have been chosen and have been utilized some typical oleaginous microorganism thalline and short chain alcohol to prepare the process of long chain fatty acid ester for feedstock conversion, contribute to understand this patent, but do not limit in any form application of the present invention.
Embodiment 1
According to document (Li Yonghong, Liu Bo, Zhao Zongbao, Bai Fengwu. biotechnology journal, 2006,22,650) described method, batch fermentation is cultivated oleaginous yeast R.toruloids AS2.1389(bacterium source in Chinese common micro-organisms culture presevation administrative center), the density that obtains fermenting is the fermentation liquid of 18.3g dry mycelium (CDW)/L, oil quantity is 13.9g/L, thalline fat content 75.9%.Get above-mentioned fermentation liquid 6mL, add ethanol, the final volume percentage concentration that makes ethanol is 10%, mix, adjust pH to 4 with 5M sodium hydroxide solution, in system, the mass percentage concentration of water is 88%, reaction solution stirs 48h under 30 ° of C, then add isopyknic granulated glass sphere, concuss 30 minutes, adds 10mL chloroform fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 90%.In product, more than 95% be that carbon chain lengths is 18 and 20 long-chain fat acetoacetic ester, can be used as biofuel.
Embodiment 2
Other operational conditions are with embodiment 1, but need the centrifugal fermentation liquid wet thallus that obtains before reaction.In wet thallus, add second alcohol and water, the final volume percentage concentration that makes ethanol is 10%, and the mass percentage concentration of water is 88%, and the yield of long-chain fat acetoacetic ester is 91%.
Embodiment 3
Other operational conditions are with embodiment 1, but before reaction, need centrifugal fermentation liquid and by thalline freeze-drying.In dry mycelium, add second alcohol and water, the final volume percentage concentration that makes ethanol is 10%, and the mass percentage concentration of water is 88%, and the yield of long-chain fat acetoacetic ester is 85%.
Embodiment 4
Other operational conditions are with embodiment 1, but the final volume percentage concentration of ethanol is 1%, and in system, the mass percentage concentration of water is 97%, and the yield of long-chain fat acetoacetic ester is 76%.
Embodiment 5
Other operational conditions are with embodiment 1, but the final volume percentage concentration of ethanol is 40%, and in system, the mass percentage concentration of water is 58%, and the yield of long-chain fat acetoacetic ester is 98%.
Embodiment 6
Other operational conditions are with embodiment 1, but reaction system are adjusted to pH10 with 5M sodium hydroxide, and the yield of long-chain fat acetoacetic ester is 35%.
Embodiment 7
Other operational conditions are with embodiment 1, but with 1M hydrochloric acid, reaction system are adjusted to reaction system pH2, and the yield of long-chain fat acetoacetic ester is 55%.
Embodiment 8
Other operational conditions are with embodiment 1, but temperature of reaction is 60 ° of C, and the yield of long-chain fat acetoacetic ester is 83%.
Embodiment 9
Other operational conditions are with embodiment 1, but temperature of reaction is 25 ° of C, and the yield of long-chain fat acetoacetic ester is 80%.
Embodiment 10
Other operational conditions are with embodiment 1, but the reaction times be 6h, the yield of long-chain fat acetoacetic ester is 60%.
Embodiment 11
Other operational conditions are with embodiment 1, but the reaction times be 168h, the yield of long-chain fat acetoacetic ester is 96%.
Embodiment 12
Other operational conditions are with embodiment 1, but short chain alcohol used is methyl alcohol, and the yield of longer chain fatty acid methyl esters is 94%.
[0027] embodiment 13
Other operational conditions are with embodiment 1, but short chain alcohol used is Virahol, and the yield of long-chain fat isopropyl propionate is 88%.
Embodiment 14
Other operational conditions are with embodiment 1, but short chain alcohol used is propyl carbinol, and the yield of the positive butyl ester of longer chain fatty acid is 80%.
Embodiment 15
Other operational conditions are with embodiment 1, but short chain alcohol used is the mixture of methyl alcohol and ethanol, the final volume percentage concentration of methyl alcohol and ethanol is 5% and 5%, and in system, the mass percentage concentration of water is 89%, and the total yield of longer chain fatty acid methyl esters and long-chain fat acetoacetic ester is 86%.
Embodiment 16
Other operational conditions are with embodiment 1, but short chain alcohol used is the fermentation liquid that contains ethanol, its preparation method reference (Yuan WJ, Chang BL, Ren JG, Liu JP, Bai FW, Li YY.Journal of AppliedMicrobiology, 2012,112,38), the final volume percentage concentration of ethanol is 9%, and in system, the mass percentage concentration of water is 89%, and the yield of long-chain fat acetoacetic ester is 85%.
Embodiment 17
Other operational conditions are with embodiment 1, but short chain alcohol used is the fermentation liquid that contains propyl carbinol, its preparation method reference (Shen CR, Lan EI, DekishimaY, Baez A, Cho KM, Liao JC.Applied andEnvironmental Microbiology, 2011,77,2905), the final volume percentage concentration of propyl carbinol is 2.5%, and in system, the mass percentage concentration of water is 96%, and the yield of the positive butyl ester of longer chain fatty acid is 67%.
Embodiment 18
Other operational conditions are with embodiment 1, but short chain alcohol used is the fermentation liquid that simultaneously contains propyl carbinol and ethanol, its preparation method reference (Xue C, Zhao JB, Lu CC, Yang ST, Bai FW, Tang IC.Biotechnology and Bioengineering, 2012,109,2746), the final volume percentage concentration of propyl carbinol and ethanol is respectively 8% and 0.7%, and in system, the mass percentage concentration of water is 89%, and the total yield of the positive butyl ester of longer chain fatty acid and long-chain fat acetoacetic ester is 83%.
Embodiment 19
Other operational conditions are with embodiment 1, but reaction solution stirs after 48h under 30 ° of C, do not carry out cytoclasis, directly add 20mL chloroform fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 71%.
Embodiment 20
Other operational conditions are with embodiment 1, but reaction solution stirs after 48h under 30 ° of C, adding concentrated hydrochloric acid to hydrochloric acid final concentration is 4M, 80 ° of C water-bath 1h, are cooled to 25 ° of C, add 10mL methylene dichloride fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 89%.
Embodiment 21
Other operational conditions are with embodiment 1, but reaction solution stirs after 48h under 30 ° of C, and under atmospheric pressure microwave (800W) is processed 1 minute, be cooled to 25 ° of C, add β-1,3-mannase to final concentration is 0.4g/L, under pH4.5,25 ° of C, processes 1h, add 20mL ethyl acetate, under 25 ° of C, fully mix, lixiviate 1h, organic phase is taken out in centrifugation, evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 84%.
Embodiment 22
Other operational conditions are with embodiment 1, but reaction solution stirs after 48h under 30 ° of C, under 121 ° of C, process 1h, be cooled to 25 ° of C, add 20mL sherwood oil fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 81%.
Embodiment 23
According to document (Hongwei Shen, Jin Guojie, Hu Cuimin, Gong Zhiwei, Bai Fengwu, Zhao Zongbao. biotechnology journal, 2012,28,56) described method, continuously ferment and cultivate oleaginous yeast R.toruloides AS2.1389(bacterium source in Chinese common micro-organisms culture presevation administrative center), the density that obtains fermenting is the fermentation liquid of 1.8g dry mycelium (CDW)/L, oil quantity is 0.6g/L.Get above-mentioned fermentation liquid 6mL, add ethanol, the final volume percentage concentration that makes ethanol is 10%, mix, adjust pH to 4 with 5M sodium hydroxide solution, in system, the mass percentage concentration of water is 90%, reaction solution stirs 168h under 20 ° of C, then add isopyknic granulated glass sphere, concuss 30 minutes, adds 10mL chloroform fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 95%.
Embodiment 24
According to document (Lin JT, Shen HW, Tan HD, Zhao X, Wu SG, Hu CM, Zhao ZB.Journal ofBiotechnology, 2011,152,184) described method, two stage fermentations are cultivated oleaginous yeast L.stakeyiAS 2.1560(bacterium sources in Chinese common micro-organisms culture presevation administrative center), the density that obtains fermenting is the fermentation liquid of 104.6g dry mycelium (CDW)/L, and oil quantity is 67.9g/L.Get above-mentioned fermentation liquid 100mL, add ethanol, making the final volume percentage concentration of ethanol is 10%, mixes, and adjusts pH to 4 with 5M sodium hydroxide solution, and in system, the mass percentage concentration of water is 80%, and reaction solution stirs 96h under 35 ° of C; Add 100mL chloroform, with 5M hydrochloric acid tune pH to 2.5, lixiviate 1h under 55 ° of C, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 90%.
Embodiment 25
According to document (Xiong W, Gao CF, Yan D, Wu C, Wu Q Y.Bioresource Technology, 2010,101,2287) described method, light autotrophy is cultivated oil-producing microalgae C.protothecoides CS-41(bacterium source in the micro-algae of Australian CSIRO research centre), the density that obtains fermenting is the fermentation liquid of 0.6g dry mycelium (CDW)/L, and oil quantity is 0.2g/L.Get above-mentioned fermentation liquid 6mL, add ethanol, the final volume percentage concentration that makes ethanol is 10%, mix, adjust pH to 4 with 5M sodium hydroxide solution, in system, the mass percentage concentration of water is 90%, reaction solution stirs 96h under 30 ° of C, then add isopyknic granulated glass sphere, concuss 30 minutes, adds 10mL chloroform fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 94%.
Embodiment 26
According to document (Xiong W, Gao CF, Yan D, Wu C, Wu Q Y.Bioresource Technology, 2010,101,2287) described method, after first light autotrophy, the mixotrophism mode of heterotrophism is cultivated oil-producing microalgae C.protothecoides CS-41(bacterium source in the micro-algae of Australian CSIRO research centre), the density that obtains fermenting is the fermentation liquid of 32g dry mycelium (CDW)/L, and oil quantity is 14g/L.Get above-mentioned fermentation liquid 6mL, add ethanol, the final volume percentage concentration that makes ethanol is 10%, mix, adjust pH to 6 with 5M sodium hydroxide solution, in system, the mass percentage concentration of water is 87%, reaction solution stirs 48h under 45 ° of C, then add isopyknic granulated glass sphere, concuss 30 minutes, adds 10mL chloroform fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 92%.
Embodiment 27
According to document (Xue FY, Miao JX, Zhang X, Tan TW.Applied Biochemistry andBiotechnology, 2010,160,498) described method, mixed culture oleaginous yeast R.glutinis AS2.703(bacterium source is in Chinese common micro-organisms culture presevation administrative center) and oil-producing microalgae C.protothecoides CS-41(bacterium source in the micro-algae of Australian CSIRO research centre), the density that obtains fermenting is the fermentation liquid of 3.7g dry mycelium (CDW)/L, and oil quantity is 0.5g/L.Get above-mentioned fermentation liquid 6mL, add ethanol, the final volume percentage concentration that makes ethanol is 10%, mix, adjust pH to 4 with 5M sodium hydroxide solution, in system, the mass percentage concentration of water is 90%, reaction solution stirs 48h under 30 ° of C, then add isopyknic granulated glass sphere, concuss 30 minutes, adds 10mL chloroform fully to mix under 25 ° of C, lixiviate 1h, organic phase is taken out in centrifugation, and evaporating solvent obtains long-chain fat acetoacetic ester, and yield is 95%.
Embodiment 28
Get 0.5 gram, the long-chain fat acetoacetic ester sample that embodiment 24 obtains, according to document (Peng BX, Yao Y, Zhao C, Lercher JA.Angewandte Chemie International Edition2012,51,2072) described method, take 10wt%Ni/HBeta as catalyzer, be 40 bar at hydrogen pressure, under 260 ° of C of temperature, react, obtain liquid product, wherein the alkane of carbon atom number 14-20 accounts for more than 95%, can be used for preparing diesel oil.
Comparative example 1
Other operational conditions are with embodiment 1, but add isopyknic granulated glass sphere before reaction, concuss 30 minutes, and microscopic examination finds, and the percentage of damage of R.toruloides cell reaches more than 95%, and the yield of long-chain fat acetoacetic ester is 6%.Relatively the experimental result of comparative example 1 and embodiment 1, shows that the integrity of oleaginous microorganism cell is most important to preparing long chain fatty acid ester; When after cytoclasis, the transformation efficiency of long chain fatty acid ester is very low, there is no actual application value.Therefore,, while preparing long chain fatty acid ester by the present invention, need to keep the integrity of cell.Comparative example 1 also illustrates, the present invention adopts lipase, immobilized lipase with other or only expresses compared with the whole-cell catalytic system of lipase, has obvious technical superiority.
The invention has the beneficial effects as follows:
Compared with utilizing the method for catalyst production long chain fatty acid ester, the present invention can be directly take the fermentation liquid of oleaginous microorganism and the fermentation liquid that contains short chain alcohol as raw material, conversion process is strong to moisture tolerance, does not need to add extra catalyzer, free from environmental pollution;
Compared with utilizing the method for super (Asia) critical fluids production long chain fatty acid ester, operational condition gentleness of the present invention, low for equipment requirements, energy consumption is low, and cost is low, and process is simple, easily amplifies;
Compared with utilizing the method for microorganism direct production long chain fatty acid ester, the present invention does not need bacterial strain to carry out the genetic engineering modified of complexity, and process adjustment is simple, and production concentration is high;
In a word, the technology of the present invention is simply effective, and energy consumption is low, and facility investment is few, and cost is low, has improved the Technical Economy of long chain fatty acid ester, is conducive to large-scale industrial production.
Claims (9)
1. a preparation method for long chain fatty acid ester, is characterized in that: in the aqueous solution, short chain alcohol conversion reaction obtains long chain fatty acid ester by oleaginous microorganism thalline.
2. be further characterized in that in accordance with the method for claim 1: described oleaginous microorganism thalline is eukaryotic microorganisms thalline or the prokaryotic micro-organisms thalline that fat content exceedes its dry weight 20%.
3. according to the method described in claim 1 or 2, be further characterized in that: the state of described oleaginous microorganism thalline is one or more necessity combination in dry mycelium, wet thallus, fermentation liquid.
4. be further characterized in that in accordance with the method for claim 1: described short chain alcohol is one or more necessity combination in methyl alcohol, ethanol, propyl alcohol, butanols, amylalcohol.
5. according to the method described in claim 1 or 4, be further characterized in that: in described water solution system, the concentration expressed in percentage by volume of short chain alcohol is 0.1%～90%.
6. be further characterized in that in accordance with the method for claim 1: in described water solution system, the mass percentage concentration of water is 10%～99%.
7. in accordance with the method for claim 1, be further characterized in that: pH2～10 of described water solution system, 20 ° of C～60 ° C of temperature of reaction, 5 minutes～168 hours time.
8. in accordance with the method for claim 1, be further characterized in that: the long chain fatty acid ester obtaining, can be with the lower liquid state organics of polarity, as a kind of in sherwood oil, normal hexane, ethyl acetate, methylene dichloride, chloroform, No. six solvent for extraction, No. four solvents, industrial hexane etc. or their necessity combination, extract purifying.
9. be further characterized in that in accordance with the method for claim 1: described in the long chain fatty acid ester that obtains can be used for preparing long chain aliphatic alcohol, aviation fuel and other chemical.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560227A (en) * | 2014-08-05 | 2015-04-29 | 浙江大学 | Method for preparing biodiesel by utilizing microwave-assisted interesterification extraction of wet algae biomass |
| CN111057570A (en) * | 2019-11-26 | 2020-04-24 | 大连立和盈海科技有限公司 | System and method for preparing biofuel |
| CN112522034A (en) * | 2020-11-09 | 2021-03-19 | 中国林业科学研究院林产化学工业研究所 | Method for preparing biofuel from rhodosporidium toruloides |
| CN115121578A (en) * | 2022-07-01 | 2022-09-30 | 北京嘉博文生物科技有限公司 | Kitchen waste grease treatment process cooperatively treated with kitchen waste |
-
2012
- 2012-12-20 CN CN201210559238.9A patent/CN103882069B/en active Active
Non-Patent Citations (3)
| Title |
|---|
| 冯冲，等: "发酵性丝孢酵母产生油脂条件的优化研究", 《河南农业大学学报》 * |
| 冯冲，等: "发酵性丝孢酵母产生油脂条件的优化研究", 《河南农业大学学报》, vol. 43, no. 6, 31 December 2009 (2009-12-31) * |
| 李永红,等: "圆红冬孢酵母菌发酵产油脂培养基及发酵条件的优化研究", 《生物工程学报》 * |
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