CN1923960A - Microorganism fermentation grease and method for preparing organism diesel oil from the same - Google Patents
Microorganism fermentation grease and method for preparing organism diesel oil from the same Download PDFInfo
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- CN1923960A CN1923960A CNA200610113582XA CN200610113582A CN1923960A CN 1923960 A CN1923960 A CN 1923960A CN A200610113582X A CNA200610113582X A CN A200610113582XA CN 200610113582 A CN200610113582 A CN 200610113582A CN 1923960 A CN1923960 A CN 1923960A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 70
- 230000004151 fermentation Effects 0.000 title claims abstract description 70
- 238000000034 method Methods 0.000 title claims abstract description 30
- 244000005700 microbiome Species 0.000 title claims abstract description 10
- 239000004519 grease Substances 0.000 title claims description 35
- 239000002283 diesel fuel Substances 0.000 title 1
- 230000000813 microbial effect Effects 0.000 claims abstract description 76
- 239000003225 biodiesel Substances 0.000 claims abstract description 23
- 239000002994 raw material Substances 0.000 claims abstract description 20
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 17
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol group Chemical group OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 17
- 238000006243 chemical reaction Methods 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 10
- 238000005809 transesterification reaction Methods 0.000 claims abstract description 9
- 239000006227 byproduct Substances 0.000 claims abstract description 8
- 239000003054 catalyst Substances 0.000 claims abstract description 7
- 238000000605 extraction Methods 0.000 claims abstract description 6
- 230000009471 action Effects 0.000 claims abstract description 4
- 238000000926 separation method Methods 0.000 claims abstract description 4
- 239000002551 biofuel Substances 0.000 claims description 31
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- 150000002632 lipids Chemical class 0.000 claims description 20
- HEDRZPFGACZZDS-UHFFFAOYSA-N chloroform Substances ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 19
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 13
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 12
- 239000002253 acid Substances 0.000 claims description 10
- 230000001580 bacterial effect Effects 0.000 claims description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 9
- 239000008103 glucose Substances 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 9
- 239000003513 alkali Substances 0.000 claims description 7
- 239000000306 component Substances 0.000 claims description 7
- 235000011187 glycerol Nutrition 0.000 claims description 7
- 239000002028 Biomass Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 238000011218 seed culture Methods 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 5
- 239000012533 medium component Substances 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 3
- 108090001060 Lipase Proteins 0.000 claims description 3
- 239000004367 Lipase Substances 0.000 claims description 3
- 102000004882 Lipase Human genes 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- 150000001720 carbohydrates Chemical class 0.000 claims description 3
- 235000014633 carbohydrates Nutrition 0.000 claims description 3
- 235000019387 fatty acid methyl ester Nutrition 0.000 claims description 3
- 235000019421 lipase Nutrition 0.000 claims description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000011973 solid acid Substances 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 239000002351 wastewater Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 229920001817 Agar Polymers 0.000 claims description 2
- 241000223233 Cutaneotrichosporon cutaneum Species 0.000 claims description 2
- 108010048733 Lipozyme Proteins 0.000 claims description 2
- 108010084311 Novozyme 435 Proteins 0.000 claims description 2
- 239000001888 Peptone Substances 0.000 claims description 2
- 108010080698 Peptones Proteins 0.000 claims description 2
- 241000223253 Rhodotorula glutinis Species 0.000 claims description 2
- 241000223230 Trichosporon Species 0.000 claims description 2
- 230000004913 activation Effects 0.000 claims description 2
- 239000008272 agar Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 claims description 2
- 239000000413 hydrolysate Substances 0.000 claims description 2
- FCCDDURTIIUXBY-UHFFFAOYSA-N lipoamide Chemical compound NC(=O)CCCCC1CCSS1 FCCDDURTIIUXBY-UHFFFAOYSA-N 0.000 claims description 2
- 235000019319 peptone Nutrition 0.000 claims description 2
- 238000012545 processing Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 238000012546 transfer Methods 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 abstract description 5
- 238000005119 centrifugation Methods 0.000 abstract description 4
- 239000003921 oil Substances 0.000 description 30
- 235000019198 oils Nutrition 0.000 description 30
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 230000008569 process Effects 0.000 description 5
- 235000013311 vegetables Nutrition 0.000 description 5
- 239000000047 product Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 235000019197 fats Nutrition 0.000 description 3
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 3
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 2
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 2
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 2
- 108090001030 Lipoproteins Proteins 0.000 description 2
- 102000004895 Lipoproteins Human genes 0.000 description 2
- 239000005642 Oleic acid Substances 0.000 description 2
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 2
- 235000021314 Palmitic acid Nutrition 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 238000007171 acid catalysis Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 2
- 235000020778 linoleic acid Nutrition 0.000 description 2
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- 229940098695 palmitic acid Drugs 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 239000008158 vegetable oil Substances 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000219000 Populus Species 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 238000005815 base catalysis Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000002485 combustion reaction Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000002803 fossil fuel Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 235000021313 oleic acid Nutrition 0.000 description 1
- 239000010773 plant oil Substances 0.000 description 1
- CASUWPDYGGAUQV-UHFFFAOYSA-M potassium;methanol;hydroxide Chemical compound [OH-].[K+].OC CASUWPDYGGAUQV-UHFFFAOYSA-M 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- -1 send out Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P30/00—Technologies relating to oil refining and petrochemical industry
- Y02P30/20—Technologies relating to oil refining and petrochemical industry using bio-feedstock
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Liquid Carbonaceous Fuels (AREA)
Abstract
本发明属于生物能源和生物催化技术领域，涉及一种利用微生物发酵油脂制备生物柴油的方法。包括微生物发酵油脂的制备和利用微生物发酵油脂制备生物柴油的方法，即以酵母菌为微生物产油脂菌种，通过在产油脂菌种培养基中接入该酵母菌微生物，在一定的培养条件下进行微生物发酵制备油脂，然后经离心、分离、提取得到微生物发酵油脂，以该微生物油脂为原料，在催化剂作用下进行转酯化反应，得到生物柴油，副产物为甘油。该技术反应条件温和、操作简单、易于控制、成本较低，以微生物发酵油脂为原料，具有良好的工业化应用前景。The invention belongs to the technical field of bioenergy and biocatalysis, and relates to a method for preparing biodiesel by using microorganisms to ferment oil. Including the preparation of microbial fermented oil and the method of using microbial fermented oil to prepare biodiesel, that is, using yeast as the microbial oil-producing strain, inserting the yeast microorganism into the oil-producing strain medium, under certain culture conditions Microbial fermentation is carried out to prepare oil, and then the microbial fermented oil is obtained through centrifugation, separation, and extraction. The microbial oil is used as a raw material to carry out transesterification reaction under the action of a catalyst to obtain biodiesel, and the by-product is glycerol. The technology has mild reaction conditions, simple operation, easy control, and low cost. It uses microbial fermentation oil as raw material and has good industrial application prospects.
Description
Technical field
The invention belongs to bioenergy and biocatalysis technology field, particularly a kind of microbial fermentation grease and be used to prepare method of bio-diesel oil.
Background technology
Current; Increase of population makes the contradiction of ever-increasing grease demand and natural resources critical shortage be becoming increasingly acute; particularly along with the global energy shortage and the environmental degradation that are on the rise; make people have to from the angle of environment protection and development of resources; active development substitutes the renewable new forms of energy of fossil fuel, is exactly a kind of have develop on a large scale very much potentiality and eco-friendly renewable and clean energy resource as biofuel (fatty acid methyl ester).The desired premium properties of liquid energy product such as biofuel has that energy density, sulphur content are low, sufficient combustion, lubricity are good.Since 1988, many European countries just begin the substitute of biofuel as conventional diesel, but because higher raw materials cost, make the price of biofuel be higher than conventional diesel, this has seriously restricted the industrialization process of biofuel, therefore choose suitable, oil resource is the direction of biofuel industry development cheaply.
The research of at present relevant biodiesel manufacture concentrates on the vegetables oil aspect basically, as Liu Chuanwu, poplar is steamed, Dai Xiaoan, Li Jianhong etc. are at document 1 " Henan chemical industry, 2,005 22 (6): 19～21 " " vegetables oil prepares the research of biofuel ", and Yang Zhenqiang, Xie Wenlei, Li Haitaos etc. are at document 2 " grain and grease, 2006, (3): 13～15 " " acid catalysis stearic permutoid reaction progress ", and Yao Yaguang, the discipline prestige, Fu Taijun, Zhang Chuanlong etc. are in document 3 " grain and grease, 2005, (10): 20～22 " acid catalysis sewer oil and the research of alcohols esterification " ", reported with the Vegetable oil lipoprotein to be the raw material production biofuel, but these reports all are to be raw material with vegetables oil or food and drink waste grease, utilize vegetables oil to be raw material, the cost height, and be raw material with the food and drink waste grease, also exist raw material to be difficult to the difficulty of collecting and transporting, so these reports all are difficult to realize industrial applications.Utilizing Vegetable oil lipoprotein to prepare in the process of biofuel, raw materials cost has just accounted for 70%～85% of total cost of production, and this has seriously restricted the industrial applications of biofuel.Yet some microbial cells contain a large amount of greases, prepare microbial oil by microbial fermentation, this preparation that not only can be biofuel provides cheap more raw material, and microorganism to have cell proliferation fast, with short production cycle, the required abundant raw material of growing, advantages such as low price, utilize microbial fermentation to produce grease simultaneously, the labor force more required than the agriculture production grease lacks, and is not subjected to season simultaneously, it is grease that the restriction of climate change, some oleaginous microorganisms can also transform five-carbon sugar, this characteristic is particularly useful for the utilization to lignocellulose hydrolysis and saccharification liquid, so microbial oil is the good raw material of production biofuel.The common method of production biofuel is an ester-interchange method, promptly in grease, add a certain amount of low-carbon alcohol, as methyl alcohol or ethanol etc., under the sour or biological fatty enzyme catalyst effect of alkali, generate biofuel in certain temperature and time reaction, by product is a glycerine simultaneously.The technology that the present invention produces microbial fermentation microbial oil combines with the technology that transesterification reaction prepares biofuel, has proposed a kind of microbial fermentation grease and has been used to prepare method of bio-diesel oil, has had not yet to see report.
Operating process of the present invention is simple, reaction conditions is gentle, is easy to control, cost is lower, simultaneously, utilizes the microbial fermentation preparing biodiesel from lipid not have and strives grain with the people, do not strive ground with grain, do not strive oily advantage with the people, has the favorable industrial application prospect.
Summary of the invention
The purpose of this invention is to provide a kind of method of utilizing the microbial fermentation preparing biodiesel from lipid; It is characterized in that, comprise the greasy preparation of microbial fermentation and utilize the method for microbial fermentation preparing biodiesel from lipid, it promptly is production by biological grease bacterial classification with the yeast, by in produce oil fat fermention medium, inserting this yeast microorganism, under the certain culture condition, carry out microbial fermentation and prepare grease, after centrifugal, separation, extraction, obtain the microbial fermentation grease, with this microbial oil is raw material, under catalyst action, carry out transesterification, obtain biofuel and by-product glycerin.
The described step of utilizing the microbial fermentation preparing biodiesel from lipid:
1) the greasy preparation of microbial fermentation
(1) with yeast (as S.cerevisiae yeast saccharomyces cerevisiae, R.glutinis rhodotorula glutinis, Trichosporoncutaneum skin shape trichosporon, buy from Institute of Microorganism, Academia Sinica) be the greasy bacterial classification of microbial fermentation, the activation of in slant medium, transferring, in seed culture medium, carry out seed culture then, the bacterial classification of 5-15% transfer again in fermention medium, in temperature is after 20-40 ℃, rotating speed are to cultivate 10-100h under the 100-200r/min, obtain containing the thalline of microbial oil, above-mentioned each substratum all 110 ℃, the sterilization 20 minutes;
(2) the greasy extraction of microbial fermentation: step (1) microbial fermentation is obtained a large amount of thalline, centrifugal, separated and collected thalline, and utilize methyl alcohol-chloroform method from thalline, to extract grease, operational condition is: add 5-100ml 1-7MHCl by every gram thalline, in 30-100 ℃ of water bath processing 0.5-5.0h, the cooling back adds and added methyl alcohol, chloroform extracted solution in 1: 1 by volume, centrifugal layering behind the thorough mixing, collect chloroform layer, the chloroform that adds equivalent again, repeat aforementioned operation, the chloroform in the extracting solution is reclaimed, obtain microbial oil.
2) microbial fermentation grease transesterificationization prepares biofuel: get microbial fermentation grease 5-25g, methyl alcohol with 1.0-30g is made esterifying agent, with the greasy 1～10wt%KOH of microbial fermentation is catalyzer, at 30-100 ℃, reacted 0.5-5 hour, after carrying out transesterification, obtaining fatty acid methyl ester through separation is biofuel, and by product is a glycerine.
Described medium component (weight ratio) comprising: the slant culture based component is yeast powder 0.1-5%; Peptone 0.5-6%; Glucose 1-10%; Agar 1-5%, pH=4-5.5; The seed culture based component is glucose 1-15%; (NH
4)
2SO
40.1-6%; Yeast powder 0.1-5%; KH
2PO
40.1-8%; Na
2HPO
40.1-5%; MgSO
40.1-5%, pH=4-5.5; The fermentation culture based component is glucose l-20%; (NH
4)
2SO
40.1-5%; Yeast powder 0.1-5%; KH
2PO
40.1-8%; Na
2HPO
40.1-5%; MgSO
40.1-5%; CaCl
20.01-4%; FeCl
30.01-6%, pH=4-5.5;
Described microbial fermentation grease bacterial classification also utilizes glucose, lignocellulose biomass by hydrolyzation liquid, starch based biomass hydrolysate or contains carbohydrate waste water and carries out fermentation culture for carbon source and obtain; Medium component and fermentation condition and fermentation unit are had no particular limits.
The also useful alkali of described catalyzer, acid, solid alkali, solid acid or biological lipase (as Novozym 435 and Lipozyme, deriving from Novozymes Company and Denmark Novo company respectively) carry out transesterification and prepare biofuel; Selected reaction conditions, temperature and time has different parameter areas for different catalyzer.
The invention has the beneficial effects as follows: utilize the microbial fermentation preparing biodiesel from lipid to enrich the raw material of preparation biofuel, low cost, environmental friendliness, reaction conditions gentleness, process be easy to control, be easy to industrial applications.
Embodiment
The purpose of this invention is to provide a kind of method of utilizing the microbial fermentation preparing biodiesel from lipid.Comprise the greasy preparation of microbial fermentation and utilize the method for microbial fermentation preparing biodiesel from lipid.
Principle of the present invention is to utilize produce oil fat microorganism to prepare microbial oil, through centrifugal, separate, obtain the microbial fermentation grease after the extraction, and be raw material with this microbial fermentation grease, under catalyst action, carry out transesterification, obtain biofuel, so not only expanded the raw material of biofuel, and reduced cost microbial oil fat raw material provided by the invention and also can be used for other alkali, acid, send out, solid alkali, solid acid, biological lipase etc. carries out the process that transesterificationization prepares biofuel as catalyzer, selected reaction conditions has different parameter areas as temperature and time for different catalyzer.
Microbial fermentation grease bacterial classification provided by the invention can utilize glucose, lignocellulose biomass by hydrolyzation liquid, starch based biomass hydrolysate, contain carbohydrate waste water etc. ferments for carbon source, and medium component and fermentation condition and fermentation unit are had no particular limits.
Greasy composition of gained microbial fermentation of the present invention and content are as shown in table 1.Table 1 is the composition and the content of the different lipid acid of microbial fermentation grease, as can be seen from Table 1, the greasy composition of microbial fermentation mainly contains different lipid acid, wherein Palmiticacid, stearic acid, oleic acid, linoleic acid content are more, its composition content and plant oil seemingly can be used as the raw material that transesterificationization prepares biofuel.
Table 1 is the composition and the content of the different lipid acid of microbial fermentation grease
| Lipid acid | Content (%) |
| C14:0 C16:0 (palmitic acid) C17:0 C18:0 (stearic acid) C18:1 (oleic acid) C18:2 (linoleic acid) C19:0 C20:0 C22:0 C24:0 | 0.76 17.80 1.31 10.27 53.02 5.29 0.45 1.02 0.51 1.30 |
Below again row embodiment the present invention is further described.
Embodiment 1
The microbe oil fermentation substratum of 100ml is placed the 500ml triangular flask, connecing 5% barms, is 30 ℃ in temperature, and shaking speed 180r/min bottom fermentation is after 60 hours, remove thalline after centrifugation, obtaining content is the microbial fermentation grease of 6.5g/L.
Embodiment 2
Inoculum size by 5% is inoculated in the full-automatic stirred-tank fermenter of 1L of liquid amount 70%, 30 ℃ of temperature, air flow 1vvm, the pH=6.0 of control culture environment, mixing speed 300r/min, ferment after 60 hours, remove thalline after centrifugation, obtaining content is the microbial fermentation grease of 8g/L.
Embodiment 3
Adopt methyl alcohol-chloroform method to extract microbial oil.Get microbial oil 100mg, add KOH-methanol solution 2ml and the normal hexane 2ml of 0.4mol/L, thermal agitation 2min places 15min in 40 ℃ of water-baths, add water 5ml, and standing demix is got the normal hexane layer, utilizes gas-chromatography and mass spectrometry to analyze its composition.The result is as shown in table 1 in concrete analysis.From the result that analyzes as can be seen, utilize yeast microorganism to ferment greasy main component and vegetables oil constituent class seemingly, can be used for the preparation of biofuel.
Embodiment 4
With the method for GB GB/T5530-85, measuring this greasy acid value is 25～30mgKOH/g, and the oil product acid value is too high, if directly with the preparation of base catalysis method, not only will consume too much alkaline catalysts, and can influence transformation efficiency.Needing to carry out depickling to stock oil and handle, is to mix at 1: 1 stock oil and methyl alcohol by mass ratio, and standing demix behind the stirring at normal temperature 30min is got oil reservoir, and repetition aforesaid operations 3 times makes the acid value of stock oil drop to 1～1.5mgKOH/g.
Get the stock oil 15g after the depickling, add 7.5g methyl alcohol and 1.5gKOH, at normal pressure, 60 ℃ of water-bath 1.5h, reaction finishes the centrifugal layering in back, obtains biofuel, and transformation efficiency is 91%, separates obtaining byproduct glycerine simultaneously.
Embodiment 5
With the microbial fermentation grease is raw material, and the duty ratio of Yan Zhikaifaing is that 30% modified solid catalyzer CaO/NaY is a catalyzer voluntarily, catalyst levels 3wt%, molar ratio of methanol to oil is 9: 1, in the 500mL there-necked flask, temperature of reaction is 65 ℃, stirring and refluxing reaction 3h.After reaction finished, centrifugation went out catalyzer, then filtrate is poured into standing demix in the separating funnel, and the upper strata underpressure distillation obtains the product biofuel after going out excessive methyl alcohol, and transformation efficiency is 95%, separated obtaining byproduct glycerine simultaneously.
Above-mentioned product carries out Analysis and Identification by gas-chromatography and mass spectrum.
Claims (7)
1. method of utilizing the microbial fermentation preparing biodiesel from lipid, it is characterized in that, comprise the greasy preparation of microbial fermentation and utilize the method for microbial fermentation preparing biodiesel from lipid, it promptly is production by biological grease bacterial classification with the yeast, by in produce oil fat fermention medium, inserting this yeast bacterial classification, under the certain culture condition, carry out microbial fermentation and prepare grease, through centrifugal, separate, obtain the microbial fermentation grease after the extraction, with this microbial oil is raw material, carry out transesterification under catalyst action, obtaining biofuel and by product is glycerine.
2. according to the described method of utilizing the microbial fermentation preparing biodiesel from lipid of claim 1, it is characterized in that the described step of microbial fermentation preparing biodiesel from lipid of utilizing is as follows:
1) the greasy preparation of microbial fermentation
(1) with yeast (as S.cerevisiae yeast saccharomyces cerevisiae, R.glutinis rhodotorula glutinis, Trichosporoncutaneum skin shape trichosporon, buy from Institute of Microorganism, Academia Sinica) be the greasy bacterial classification of microbial fermentation, the activation of in slant medium, transferring, in seed culture medium, carry out seed culture then, the bacterial classification of 5-15% transfer again in fermention medium, in temperature is after 20-40 ℃, rotating speed are to cultivate 10-100h under the 100-200r/min, obtain containing the thalline of microbial oil, above-mentioned each substratum all 110 ℃, the sterilization 20 minutes;
(2) the greasy extraction of microbial fermentation: step (1) microbial fermentation is obtained a large amount of thalline, centrifugal, separated and collected thalline, and utilize methyl alcohol-chloroform method from thalline, to extract grease, operational condition is: add 5-100ml 1-7MHCl by every gram thalline, in 30-100 ℃ of water bath processing 0.5-5.0h, the cooling back adds and added methyl alcohol, chloroform extracted solution in 1: 1 by volume, centrifugal layering behind the thorough mixing, collect chloroform layer, the chloroform that adds equivalent again, repeat aforementioned operation, the chloroform in the extracting solution is reclaimed, obtain microbial oil;
2) microbial fermentation grease transesterificationization prepares biofuel
Getting microbial fermentation grease 5-25g, make esterifying agent with the methyl alcohol of 1.0-30g, is catalyzer with the greasy 1～10wt%KOH of microbial fermentation, at 30-100 ℃, reacted 0.5-5 hour, carry out transesterification after, obtaining fatty acid methyl ester through separation is biofuel, and by product is a glycerine.
3. according to the described method of utilizing the microbial fermentation preparing biodiesel from lipid of claim 2, it is characterized in that described slant culture based component (weight ratio) is yeast powder 0.1-5%; Peptone 0.5-6%; Glucose 1-10%; Agar 1-5%, pH=4-5.5.
4. according to the described method of utilizing the microbial fermentation preparing biodiesel from lipid of claim 2, it is characterized in that seed culture based component (weight ratio) is glucose 1-15%; (NH
4)
2SO
40.1-6%; Yeast powder 0.1-5%; KH
2PO
40.1-8%; Na
2HPO
40.1-5%; MgSO
40.1-5%, pH=4-5.5.
5. according to the described method of utilizing the microbial fermentation preparing biodiesel from lipid of claim 2, it is characterized in that fermentation culture based component (weight ratio) is glucose 1-20%; (NH
4)
2SO
40.1-5%; Yeast powder 0.1-5%; KH
2PO
40.1-8%; Na
2HPO
40.1-5%; MgSO
40.1-5%; CaCl
20.01-4%; FeCl
30.01-6%, pH=4-5.5.
6. according to the described method of utilizing the microbial fermentation preparing biodiesel from lipid of claim 1, it is characterized in that described microbial fermentation grease bacterial classification also utilizes glucose, lignocellulose biomass by hydrolyzation liquid, starch based biomass hydrolysate or contains carbohydrate waste water and carries out fermentation culture for carbon source and obtain; Medium component and fermentation condition and fermentation unit are had no particular limits.
7. according to the described method of utilizing the microbial fermentation preparing biodiesel from lipid of claim 1, it is characterized in that, the also useful alkali of described catalyzer, acid, solid alkali, solid acid or biological lipase (as Novozym 435 and Lipozyme, deriving from Novozymes Company and Denmark Novo company respectively) carry out transesterification and prepare biofuel; Selected reaction conditions, temperature and time has different parameter areas for different catalyzer.
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