CN106119308A - A kind of method that magnetic carrier immobilized-lipase prepares biodiesel - Google Patents
A kind of method that magnetic carrier immobilized-lipase prepares biodiesel Download PDFInfo
- Publication number
- CN106119308A CN106119308A CN201610152923.8A CN201610152923A CN106119308A CN 106119308 A CN106119308 A CN 106119308A CN 201610152923 A CN201610152923 A CN 201610152923A CN 106119308 A CN106119308 A CN 106119308A
- Authority
- CN
- China
- Prior art keywords
- lipase
- magnetic
- immobilized
- sio
- magnetic nanoparticle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000004367 Lipase Substances 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 30
- 239000003225 biodiesel Substances 0.000 title claims abstract description 29
- 230000005291 magnetic effect Effects 0.000 title claims abstract description 27
- 239000002122 magnetic nanoparticle Substances 0.000 claims abstract description 33
- 239000003054 catalyst Substances 0.000 claims abstract description 30
- 102000004882 Lipase Human genes 0.000 claims abstract description 21
- 108090001060 Lipase Proteins 0.000 claims abstract description 21
- 235000019421 lipase Nutrition 0.000 claims abstract description 21
- 102000004190 Enzymes Human genes 0.000 claims abstract description 13
- 108090000790 Enzymes Proteins 0.000 claims abstract description 13
- 241000894006 Bacteria Species 0.000 claims abstract description 8
- 108010093096 Immobilized Enzymes Proteins 0.000 claims abstract description 4
- 238000004132 cross linking Methods 0.000 claims abstract description 4
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 30
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 239000003921 oil Substances 0.000 claims description 25
- 235000019198 oils Nutrition 0.000 claims description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 238000006243 chemical reaction Methods 0.000 claims description 21
- 238000002360 preparation method Methods 0.000 claims description 16
- 229910052681 coesite Inorganic materials 0.000 claims description 11
- 229910052906 cristobalite Inorganic materials 0.000 claims description 11
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
- 229910052682 stishovite Inorganic materials 0.000 claims description 11
- 238000009210 therapy by ultrasound Methods 0.000 claims description 11
- 229910052905 tridymite Inorganic materials 0.000 claims description 11
- 239000002699 waste material Substances 0.000 claims description 11
- 239000002105 nanoparticle Substances 0.000 claims description 10
- 239000012153 distilled water Substances 0.000 claims description 9
- 239000004006 olive oil Substances 0.000 claims description 9
- 235000008390 olive oil Nutrition 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 8
- 238000005576 amination reaction Methods 0.000 claims description 7
- 238000000855 fermentation Methods 0.000 claims description 7
- 230000004151 fermentation Effects 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 239000008363 phosphate buffer Substances 0.000 claims description 7
- 238000003828 vacuum filtration Methods 0.000 claims description 7
- 238000013019 agitation Methods 0.000 claims description 6
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 5
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 5
- 239000002131 composite material Substances 0.000 claims description 5
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 4
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 4
- 230000001186 cumulative effect Effects 0.000 claims description 4
- 238000004064 recycling Methods 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 3
- -1 Amino Chemical group 0.000 claims description 2
- 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 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 2
- 239000000872 buffer Substances 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 239000004519 grease Substances 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- ALSTYHKOOCGGFT-KTKRTIGZSA-N (9Z)-octadecen-1-ol Chemical compound CCCCCCCC\C=C/CCCCCCCCO ALSTYHKOOCGGFT-KTKRTIGZSA-N 0.000 claims 1
- 229940055577 oleyl alcohol Drugs 0.000 claims 1
- XMLQWXUVTXCDDL-UHFFFAOYSA-N oleyl alcohol Natural products CCCCCCC=CCCCCCCCCCCO XMLQWXUVTXCDDL-UHFFFAOYSA-N 0.000 claims 1
- 238000005809 transesterification reaction Methods 0.000 abstract description 13
- 238000006555 catalytic reaction Methods 0.000 abstract description 6
- 239000002253 acid Substances 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 108010091086 Recombinases Proteins 0.000 description 3
- 102000018120 Recombinases Human genes 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000006227 byproduct Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000001291 vacuum drying Methods 0.000 description 3
- HPEUJPJOZXNMSJ-UHFFFAOYSA-N Methyl stearate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC HPEUJPJOZXNMSJ-UHFFFAOYSA-N 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 238000005815 base catalysis Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 230000032050 esterification Effects 0.000 description 2
- 238000005886 esterification reaction Methods 0.000 description 2
- 229960004756 ethanol Drugs 0.000 description 2
- 239000003925 fat Substances 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000002440 industrial waste Substances 0.000 description 2
- 150000004702 methyl esters Chemical class 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000001149 (9Z,12Z)-octadeca-9,12-dienoate Substances 0.000 description 1
- WTTJVINHCBCLGX-UHFFFAOYSA-N (9trans,12cis)-methyl linoleate Natural products CCCCCC=CCC=CCCCCCCCC(=O)OC WTTJVINHCBCLGX-UHFFFAOYSA-N 0.000 description 1
- LNJCGNRKWOHFFV-UHFFFAOYSA-N 3-(2-hydroxyethylsulfanyl)propanenitrile Chemical compound OCCSCCC#N LNJCGNRKWOHFFV-UHFFFAOYSA-N 0.000 description 1
- FLIACVVOZYBSBS-UHFFFAOYSA-N Methyl hexadecanoate Natural products CCCCCCCCCCCCCCCC(=O)OC FLIACVVOZYBSBS-UHFFFAOYSA-N 0.000 description 1
- PKIXXJPMNDDDOS-UHFFFAOYSA-N Methyl linoleate Natural products CCCCC=CCCC=CCCCCCCCC(=O)OC PKIXXJPMNDDDOS-UHFFFAOYSA-N 0.000 description 1
- 240000007817 Olea europaea Species 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000003965 capillary gas chromatography Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000002485 combustion reaction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000002761 deinking Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- QYDYPVFESGNLHU-UHFFFAOYSA-N elaidic acid methyl ester Natural products CCCCCCCCC=CCCCCCCCC(=O)OC QYDYPVFESGNLHU-UHFFFAOYSA-N 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- CAMHHLOGFDZBBG-UHFFFAOYSA-N epoxidized methyl oleate Natural products CCCCCCCCC1OC1CCCCCCCC(=O)OC CAMHHLOGFDZBBG-UHFFFAOYSA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 235000019387 fatty acid methyl ester Nutrition 0.000 description 1
- 150000002191 fatty alcohols Chemical class 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 235000019626 lipase activity Nutrition 0.000 description 1
- 230000001050 lubricating effect Effects 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- FLIACVVOZYBSBS-GZXVCZRGSA-N methyl hexadecanoate Chemical group CCCCCCCCCCCCCCC[14C](=O)OC FLIACVVOZYBSBS-GZXVCZRGSA-N 0.000 description 1
- QYDYPVFESGNLHU-KHPPLWFESA-N methyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC QYDYPVFESGNLHU-KHPPLWFESA-N 0.000 description 1
- 229940073769 methyl oleate Drugs 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- 238000006053 organic reaction Methods 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- LFQCEHFDDXELDD-UHFFFAOYSA-N tetramethyl orthosilicate Chemical compound CO[Si](OC)(OC)OC LFQCEHFDDXELDD-UHFFFAOYSA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- C12P7/6436—Fatty acid esters
- C12P7/649—Biodiesel, i.e. fatty acid alkyl esters
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- 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
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
Abstract
The invention discloses a kind of method that magnetic carrier immobilized-lipase prepares biodiesel, the method utilizes immobilized-lipase magnetic nanoparticle catalyst to be catalyzed reaction, and described immobilized-lipase magnetic nanoparticle catalyst is used the method for covalent cross-linking to be prepared from yielding lipase being fixed of bacterium by magnetic nanoparticle as carrier.Technical solution of the present invention simple process is easy, low for equipment requirements, and the catalyst transesterification efficiency related to is high, and immobilized enzyme has enzyme and lives high and stable, it is simple to reclaims and separates, can repeatedly use, avoids using strong acid and strong base simultaneously, reduce the pollution to environment.
Description
Technical field
The invention belongs to biological engineering and derived energy chemical field, be specifically related to one and utilize magnetic nano particle immobilized fat
The method of biodiesel is prepared in fat enzyme catalysis.
Background technology
Along with economic growth and social progress, the consumption of fossil energy especially petroleum-based energy increases rapidly, develops novel
Bioenergy, as petrifaction diesel substitute products, can promote novel energy development, ensure energy security and national security.Biological bavin
Oil not only has with the similar combustion characteristics of petrifaction diesel, also has environmental friendliness, higher-security, good lubricating properties and can be again
Life is biodegradable waits good characteristic.
In each production technology of biodiesel, chemical bases catalysis method has realized industrialized production, although its catalyst is the most honest and the cleanest
Valency, response time are short, the advantage that conversion ratio is higher, but still suffer from a series of intrinsic defect: material quality is required relatively by (1)
Height is low to waste oil, industrial waste recovery utilization rate；(2) complex process, energy consumption is high；(3) alcohol consumption is big and is not easily recycled profit
With；(4) by-product glycerin difficulty recycles, and catalyst difficulty separates with product, increases the cost of subsequent treatment, and spent lye discharges
Form new environomental pollution source.Base catalysis method utilizes refined oil with methanol-fueled CLC biodiesel, although alleviate to a certain extent
Pressure on Energy, but it is not energy-conserving and environment-protective that the inherent shortcoming of this technique determines it, therefore have more environment-friendly advantage
The trend of green energy resource technique lipase-catalyzed technique necessarily Future Development.
Lipase-catalyzed method is low to the selectivity of raw material, waste oil that available acid number is higher and industrial waste, and reaction
Mild condition, energy consumption is low；Alcohol consumption is little, product and by-product is more easily separated and non-pollution discharge, overcomes the tight of base catalysis method
Weight defect, is a kind of preferable energy saving and environment friendly technique.But, lipase-catalyzed chemical reaction process condition, it is common that
Various organic solvent mediums (short chain alcohol as used in production of biodiesel), extreme soda acid (strong as in deinking industry
Alkalescence) etc. under the conditions of, be easily caused the inactivation of native lipase, thus limit lipase large-scale application, lipase steady
Qualitative difference is mainly reflected in resolvase and is directly exposed in reaction system, and methanol or ethanol prepare biodiesel as acyl acceptor
Time, poor with the compatibility of oils and fats, react the desired amount of methanol or the undissolved part of ethanol will cause enzyme to inactivate.
Biodiesel preparation technology is had a major impact by the preparation method of lipase.Resolvase is easy in organic reaction system
Producing and assemble, be unfavorable for contacting of enzyme and reactant, reaction efficiency is the lowest, poor stability.The short chain fatty alcohols such as reactant methanol
Effect toxic to enzyme with by-product glycerin, reduces service life and the conversion ratio of enzyme.Substantial amounts of resolvase cannot obtain many
Secondary utilization, reclaims difficulty, and the price of lipase itself is the most sufficiently expensive in addition so that the cost of Production by Enzymes biodiesel is very
High, it is impossible to for promoting the use of of industry.Lose little by physisorphtion lipase, and remain the specificity of lipase,
But the poor stability of immobilized-lipase, is difficult to storage；The loss of cross-linking method lipase is relatively big, but the stability of immobilized-lipase
Preferably, the vigor of 62.5% is retained after 3 months.
Summary of the invention
The problem existed for above process for preparing biodiesel by enzyme, it is contemplated that in overcoming the deficiencies in the prior art, carry
A kind of method preparing biodiesel for magnetic carrier immobilized-lipase, realizes the recycling of lipase by immobilization,
Reduce the production cost of synthesis of biodiesel by enzymatic catalysis.
In order to solve above-mentioned technical problem, the present invention is accomplished by:
A kind of method that magnetic carrier immobilized-lipase prepares biodiesel, the method utilizes immobilized-lipase magnetic
Reaction is catalyzed by nanoparticle catalyst, and described immobilized-lipase magnetic nanoparticle catalyst is by magnetic nanoparticle
Yielding lipase being fixed of bacterium is prepared from by the method using covalent cross-linking as carrier.
Described magnetic nanoparticle is prepared as steps described below:
1)Fe3O4@SiO2The preparation of magnetic composite nano particle: weigh Fe3O4Nano-particle is put into containing dehydrated alcohol, steaming
In the there-necked flask of distilled water and ammonia water mixture, after ultrasonic Treatment 1h, it is maintained under 30 DEG C of constant temperature and carries out mechanical agitation
30min, dropping tetraethyl orthosilicate (TEOS), then mechanic whirl-nett reaction 24h, carry out Magneto separate with Magnet, use distilled water the most again
Fully washing with dehydrated alcohol, Magneto separate, vacuum filtration is dried to mass conservation, obtains loose powder Fe3O4@SiO2
Magnetic composite nano particle；
2)Fe3O4@SiO2-NH2The preparation of amination magnetic nanoparticle: weigh step 1) the middle Fe prepared3O4@SiO2Magnetic
Property composite nanometer particle put in the there-necked flask containing dehydrated alcohol and ammonia water mixture, after ultrasonic Treatment 1h, protect
Hold under 30 DEG C of constant temperature, drip gamma-aminopropyl-triethoxy-silane (APTES), mechanic whirl-nett reaction 24h, carry out magnetic with Magnet and divide
From, fully washing with distilled water and dehydrated alcohol the most respectively, Magneto separate, vacuum filtration is dried to mass conservation, obtains loose
Powder Fe dissipated3O4@SiO2–NH2Amination magnetic nanoparticle；
3)Fe3O4@SiO2The preparation of-CHO activation magnetic nanoparticle: weigh step 2 again) Fe for preparing3O4@SiO2-NH2
Amination magnetic Nano is put in the there-necked flask containing glutaraldehyde solution, after ultrasonic Treatment 30min, is maintained at 30
Mechanical agitation 12h under DEG C constant temperature, carries out Magneto separate with Magnet, and recycling phosphate buffer fully washs, and removes unnecessary penta 2
Aldehyde, vacuum filtration is dried to mass conservation, obtains loose powder Fe3O4@SiO2-CHO activates magnetic nanoparticle.
The preparation of described immobilized-lipase magnetic nanoparticle catalyst, specifically includes following steps:
1) yielding lipase is cultivated in following condition: the glucose of 2.845g/L, the olive oil addition of 1.2%,
Initial pH is the Carnis Bovis seu Bubali cream, (NH of 0.1% of 7.2,1.5%4)2SO4, the K of inoculum concentration 2.5% and 0.15%2HPO4.3H2O, protects
Holding under 30 DEG C of constant temperature, ferment 48h；
2) by step 1) in cultured fermentation enzyme bacterium solution centrifugal 10min under 12000rpm rotating speed, collect supernatant, will
Fe3O4@SiO2-CHO activation magnetic nanoparticle, the phosphate buffer of pH7.0 and fermentation enzyme bacterium solution are mixed in triangular flask, logical
Cross ultrasonic Treatment, and shaking table reaction being fixed of 4h under conditions of 40 DEG C, then carry out Magneto separate with Magnet, use phosphoric acid
Buffer fully washs, vacuum filtration drying to obtain immobilized-lipase magnetic nanoparticle catalyst.
When the present invention utilizes magnetic immobilized enzyme biodiesel, add alcohol according to mol ratio 4:1-12:1
The immobilization of oil, the water adding the 10-15% accounting for oil alcohol and water cumulative volume and the 2-8% accounting for oil, alcohol, water and catalyst gross mass
Lipase magnetic nanoparticle catalyst, is placed in the shaking table that reaction temperature is 30-50 DEG C and carries out reacting 4h.Described grease separation olive
Olive oil or waste oil；Described alcohol is methanol.
Compared with prior art, the invention have the benefit that high yield lipase strain used in the present invention is through
After the fermentation of excellent fermentation condition, institute's yielding lipase activity is higher, and immobilization technology is fixed lipase catalyzed transesterification with traditional
Preparation biodiesel is compared, and the immobilized-lipase magnetic nanoparticle catalyst that the present invention provides directly utilizes microorganism and sends out
Ferment liquid, it is not necessary to carry out the purification of lipase, technique is simple, with low cost；Magnetic material is selected to be easy to separate, it is achieved to be repeated several times
Utilize, after esterification terminates, utilize permanent magnet to be separated from product by catalyst, can directly be invested in new anti-
Answer in thing and react；There is high service life, mass-transfer efficiency and conversion ratio: owing to catalyst has magnetics effect, therefore
Can put into have in the reactor in magnetic field by catalyst and react, catalyst distribution so can be made more uniform, reduce
Sedimentation collision, reaches to improve mass-transfer efficiency and the effect of conversion ratio, transesterification reaction mild condition, and the response time is short, transesterification efficiency
High.
Accompanying drawing explanation
Fig. 1 is the immobilized-lipase magnetic nanoparticle catalyst stereoscan photograph of the present invention.
Fig. 2 is that example 1 uses olive oil to prepare biodiesel GC-MS chromatogram.
Fig. 3 is that example 2 uses Preparation of biodiesel from waste oils GC-MS chromatogram.
Detailed description of the invention
Below in conjunction with specific embodiment, the detailed description of the invention of the present invention is described in further detail, but the present invention
Content is not limited only to the content involved by embodiment.
Weigh the Fe of 2g3O4Nano-particle is put into containing 200ml dehydrated alcohol, 50ml distilled water and ammonia 5ml water mixed liquid
In there-necked flask, after ultrasonic Treatment 1h, it is maintained under 30 DEG C of constant temperature and carries out mechanical agitation 30min, be slowly added to 5ml's
Tetraethyl orthosilicate, continuous stirring reaction 24h, Magneto separate, fully wash with distilled water and dehydrated alcohol the most again, magnetic divides
From, vacuum drying, obtain loose powder Fe3O4@SiO2；Weigh the Fe of 2g3O4@SiO2Put into containing 150ml dehydrated alcohol and
In the there-necked flask of 15ml ammonia water mixture, ultrasound wave disperses, and is maintained at γ-aminopropyl three second dripping 10ml under 30 DEG C of constant temperature
TMOS, mechanic whirl-nett reaction 24h, Magneto separate, fully washs with distilled water and dehydrated alcohol, Magneto separate, very respectively
Empty dry, obtain loose powder Fe3O4@SiO2–NH2Amination magnetic nanoparticle；Weigh the Fe of 2g again3O4@SiO2-NH2Ammonia
Base magnetic Nano is put into containing in the there-necked flask of 10% glutaraldehyde solution of 100ml, by ultrasonic Treatment 30min
After, it being maintained at mechanical agitation 12h under 30 DEG C of constant temperature, Magneto separate, recycling phosphate buffer fully washs, and removes unnecessary penta 2
Aldehyde, vacuum drying, obtain loose powder Fe3O4@SiO2-CHO activates magnetic nanoparticle, and sealing saves backup.
The Fe3O4@SiO2 CHO weighing 50mg again activates magnetic nanoparticle, is the pH7.0 of 1mol/L by 20ml concentration
Phosphate buffer and the fermentation enzyme bacterium solution of 30ml be mixed in triangular flask, by ultrasonic Treatment, and under conditions of 40 DEG C
Shaking table reaction being fixed of 4h, then carries out Magneto separate with Magnet, fully washs with phosphate buffer, and vacuum drying i.e. obtains
Immobilized-lipase magnetic nanoparticle catalyst.
Embodiment 1 is catalyzed olive oil transesterification preparation biodiesel
With olive oil as raw material, carry out transesterification test with the immobilized-lipase magnetic nanoparticle catalyst of preparation.
2ml olive oil and 330ml methanol (molar ratio of methanol to oil 4:1) mixture being loaded in test tube, the water adding 10% (accounts for oil alcohol and water
The 10% of cumulative volume), then be separately added into 5% above-mentioned catalyst (account for oil, alcohol, water, the 5% of catalyst gross mass), be placed in and shake
Reacting in Chuan, temperature 40 DEG C, rotating speed 160rpm carries out reacting 4h.After having reacted, taking out sample at rotating speed is
10000rpm takes supernatant liquid and measures methyl ester content after being centrifuged 2min, recording transesterification efficiency is 98.84%.
Embodiment 2 is catalyzed waste oil transesterification preparation biodiesel
With waste oil as raw material, carry out transesterification test with the immobilized-lipase magnetic nanoparticle catalyst of preparation.
Waste oil raw material is provided by Chengdu Shu Feng company limited, through mechanical purifying, bentone adsorbent adsorption filtration, washing and high speed
After rotating centrifugal removes moisture, directly use after carrying out deacidification with methanol esterification.Waste oil key property is as follows: acid number
28.87mgKOH/g；Free fatty 10.27%；Mean molecule quantity is 885.68.By waste oil and methanol (molar ratio of methanol to oil
During 12:1) mixture is loaded on test tube, add the water the 15% of alcohol and water cumulative volume (account for oil) of 15%, then be separately added into 8% upper
State catalyst (account for oil, alcohol, water, the 8% of catalyst gross mass), be placed in shaking table and react, temperature 50 C, rotating speed 160rpm
Carry out reacting 4h.The sample taken out is to take supernatant liquid after 10000rpm is centrifuged 2min to measure methyl ester content at rotating speed, records and turns
Ester efficiency is 97.19%.
In the present invention, with this amination magnetic nanometer composite material after activated as carrier, by covalent coupling mode
The immobilized enzyme catalysis agent that fixing high yield lipase strain institute yielding lipase obtains, is being catalyzed olive oil and waste oil as substrate
Catalysis for preparing biodiesel oil, after 4h transesterification reaction, transesterification efficiency is up to 98.84% and 97.19% respectively.Transesterification product is main
It is methyl hexadecanoate, methyl oleate, methyl linoleate, methyl stearate, is the main component of biodiesel.
The i.e. fatty acid methyl ester yield of transesterification efficiency uses chromatography analysis, gas chromatogram operating condition: fid detector, hair
Capillary gas chromatography post HP-88, sample size 1 μ l, column flow rate 1ml/min, 10 DEG C/min of programming rate, temperature programming 60 DEG C-230
℃。
Above the present invention is done exemplary description, it should explanation, in the situation of the core without departing from the present invention
Under, any simple deformation, amendment or other those skilled in the art can not spend the equivalent of creative work equal
Fall into protection scope of the present invention.
Claims (6)
1. the method that a magnetic carrier immobilized-lipase prepares biodiesel, it is characterised in that: the method utilizes immobilization
Reaction is catalyzed by lipase magnetic nanoparticle catalyst, and described immobilized-lipase magnetic nanoparticle catalyst is by magnetic
Property nano-particle as carrier use covalent cross-linking method yielding lipase being fixed of bacterium is prepared from.
The method that a kind of magnetic carrier immobilized-lipase the most according to claim 1 prepares biodiesel, its feature exists
In: described magnetic nanoparticle is prepared as steps described below:
1)Fe3O4@SiO2The preparation of magnetic composite nano particle: weigh Fe3O4Nano-particle is put into containing dehydrated alcohol, distilled water
With in the there-necked flask of ammonia water mixture, after ultrasonic Treatment 1h, it is maintained under 30 DEG C of constant temperature and carries out mechanical agitation
30min, drips tetraethyl orthosilicate, then mechanic whirl-nett reaction 24h, carries out Magneto separate with Magnet, the most again with distilled water and anhydrous
Ethanol fully washs, Magneto separate, and vacuum filtration is dried to mass conservation, obtains loose powder Fe3O4@SiO2Magnetic is multiple
Close nano-particle；
2)Fe3O4@SiO2-NH2The preparation of amination magnetic nanoparticle: weigh step 1) the middle Fe prepared3O4@SiO2Magnetic is multiple
Close nano-particle and put in the there-necked flask containing dehydrated alcohol and ammonia water mixture, after ultrasonic Treatment 1h, be maintained at
Drip gamma-aminopropyl-triethoxy-silane, mechanic whirl-nett reaction 24h under 30 DEG C of constant temperature, carry out Magneto separate with Magnet, then distinguish
Fully washing with distilled water and dehydrated alcohol, Magneto separate, vacuum filtration is dried to mass conservation, obtains loose powder
Fe3O4@SiO2–NH2Amination magnetic nanoparticle；
3)Fe3O4@SiO2The preparation of-CHO activation magnetic nanoparticle: weigh step 2 again) Fe for preparing3O4@SiO2-NH2Amino
Change magnetic Nano and put in the there-necked flask containing glutaraldehyde solution, after ultrasonic Treatment 30min, be maintained at 30 DEG C of perseverances
Lower mechanical agitation 12h of temperature, carries out Magneto separate with Magnet, and recycling phosphate buffer fully washs, and removes unnecessary glutaraldehyde, very
Empty sucking filtration is dried to mass conservation, obtains loose powder Fe3O4@SiO2-CHO activates magnetic nanoparticle.
The method that a kind of magnetic carrier immobilized-lipase the most according to claim 1 prepares biodiesel, its feature exists
In the preparation of described immobilized-lipase magnetic nanoparticle catalyst, specifically include following steps:
1) yielding lipase is cultivated in following condition: the glucose of 2.845g/L, the olive oil addition of 1.2%, initially
PH is the Carnis Bovis seu Bubali cream, (NH of 0.1% of 7.2,1.5%4)2SO4, the K of inoculum concentration 2.5% and 0.15%2HPO4.3H2O, is maintained at
Under 30 DEG C of constant temperature, ferment 48h；
2) by step 1) in cultured fermentation enzyme bacterium solution centrifugal 10min under 12000rpm rotating speed, collect supernatant, will
Fe3O4@SiO2-CHO activation magnetic nanoparticle, the phosphate buffer of pH7.0 and fermentation enzyme bacterium solution are mixed in triangular flask, logical
Cross ultrasonic Treatment, and shaking table reaction being fixed of 4h under conditions of 40 DEG C, then carry out Magneto separate with Magnet, use phosphoric acid
Buffer fully washs, vacuum filtration drying to obtain immobilized-lipase magnetic nanoparticle catalyst.
The method that a kind of magnetic carrier immobilized-lipase the most according to claim 1 prepares biodiesel, its feature exists
In: utilize magnetic immobilized enzyme biodiesel, according to mol ratio 4:1-12:1 add alcohol oil, add account for oleyl alcohol and
The water of the 10-15% of water cumulative volume and account for the immobilized-lipase magnetic Nano of 2-8% of oil, alcohol, water and catalyst gross mass
Beaded catalyst, is placed in the shaking table that reaction temperature is 30-50 DEG C and carries out reacting 4h.
The method that a kind of magnetic carrier immobilized-lipase the most according to claim 4 prepares biodiesel, its feature exists
In: described grease separation olive oil or waste oil.
The method that a kind of magnetic carrier immobilized-lipase the most according to claim 4 prepares biodiesel, its feature exists
In: described alcohol is methanol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610152923.8A CN106119308A (en) | 2016-03-17 | 2016-03-17 | A kind of method that magnetic carrier immobilized-lipase prepares biodiesel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610152923.8A CN106119308A (en) | 2016-03-17 | 2016-03-17 | A kind of method that magnetic carrier immobilized-lipase prepares biodiesel |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106119308A true CN106119308A (en) | 2016-11-16 |
Family
ID=57270357
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610152923.8A Pending CN106119308A (en) | 2016-03-17 | 2016-03-17 | A kind of method that magnetic carrier immobilized-lipase prepares biodiesel |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106119308A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520622A (en) * | 2016-12-01 | 2017-03-22 | 湖南利尔康生物股份有限公司 | Production method for mycoprotein containing lipase |
| CN107326022A (en) * | 2017-03-29 | 2017-11-07 | 东北林业大学 | A kind of method of silanization magnetic cellulose microsphere fixed fat enzyme |
| CN108753761A (en) * | 2018-05-30 | 2018-11-06 | 北京理工大学珠海学院 | A kind of preparation method of amination modified composite magnetic nano-carrier for lipase immobilization |
| ES2695327A1 (en) * | 2017-06-28 | 2019-01-03 | Consejo Superior Investigacion | SYNTHESIS OF BIODIESEL CATALYZED BY AN IMMOBILIZED ENZYMATIC CRUDE ON MAGNETIC PARTICLES (Machine-translation by Google Translate, not legally binding) |
| CN111041014A (en) * | 2019-12-31 | 2020-04-21 | 浙江工业大学 | Magnetic immobilized lipase and application thereof in resolution of 1-methyl-3-amphetamine |
| CN112680436A (en) * | 2020-12-23 | 2021-04-20 | 陕西科技大学 | Immobilized enzyme for extracting anthocyanin from black rice and preparation and use methods thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102994491A (en) * | 2012-07-12 | 2013-03-27 | 浙江大学 | Immobilization method of Thermus lipase |
| CN103333879A (en) * | 2013-06-24 | 2013-10-02 | 中国水产科学研究院黄海水产研究所 | Method for immobilizing marine lipase Bohai Sea-9145 with magnetic Fe3O4@SiO2-NH2 nanoparticles |
| CN103400677A (en) * | 2013-06-24 | 2013-11-20 | 中国水产科学研究院黄海水产研究所 | Preparation method of magnetic Fe3O4@SiO2-NH2 nanoparticles |
| CN104404023A (en) * | 2014-11-05 | 2015-03-11 | 济南大学 | Preparation method of magnetic carrier immobilized lipase, and method for preparing biodiesel under catalysis of magnetic carrier immobilized lipase |
| CN105255853A (en) * | 2015-11-10 | 2016-01-20 | 河南工业大学 | Preparation method of magnetic hydroxyapatite immobilized lipase and application in edible fat and oil processing |
-
2016
- 2016-03-17 CN CN201610152923.8A patent/CN106119308A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102994491A (en) * | 2012-07-12 | 2013-03-27 | 浙江大学 | Immobilization method of Thermus lipase |
| CN103333879A (en) * | 2013-06-24 | 2013-10-02 | 中国水产科学研究院黄海水产研究所 | Method for immobilizing marine lipase Bohai Sea-9145 with magnetic Fe3O4@SiO2-NH2 nanoparticles |
| CN103400677A (en) * | 2013-06-24 | 2013-11-20 | 中国水产科学研究院黄海水产研究所 | Preparation method of magnetic Fe3O4@SiO2-NH2 nanoparticles |
| CN104404023A (en) * | 2014-11-05 | 2015-03-11 | 济南大学 | Preparation method of magnetic carrier immobilized lipase, and method for preparing biodiesel under catalysis of magnetic carrier immobilized lipase |
| CN105255853A (en) * | 2015-11-10 | 2016-01-20 | 河南工业大学 | Preparation method of magnetic hydroxyapatite immobilized lipase and application in edible fat and oil processing |
Non-Patent Citations (5)
| Title |
|---|
| CHIEN-HUNG LIU 等: "Biodiesel production by enzymatic transesterification catalyzed by Burkholderia lipase immobilized on hydrophobic magnetic particles", 《APPLIED ENERGY》 * |
| 冯超: "磁性纳米载体的制备及其固定化脂肪酶催化性能研究", 《万方数据》 * |
| 张大准: "磁性纳米材料改性及固定化脂肪酶的研究", 《万方数据》 * |
| 易笑生 等: "Fe3O4-SiO2载体制备和脂肪酶固定化条件优化", 《食品工业科技》 * |
| 马晓瑞 等: "Fe3O4@SiO2-CHO共价固定脂肪酶条件的响应面优化及固定化酶催化生产生物柴油", 《应用与环境生物学报》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520622A (en) * | 2016-12-01 | 2017-03-22 | 湖南利尔康生物股份有限公司 | Production method for mycoprotein containing lipase |
| CN106520622B (en) * | 2016-12-01 | 2020-08-04 | 湖南利尔康生物股份有限公司 | Production method of mycoprotein containing lipase |
| CN107326022A (en) * | 2017-03-29 | 2017-11-07 | 东北林业大学 | A kind of method of silanization magnetic cellulose microsphere fixed fat enzyme |
| ES2695327A1 (en) * | 2017-06-28 | 2019-01-03 | Consejo Superior Investigacion | SYNTHESIS OF BIODIESEL CATALYZED BY AN IMMOBILIZED ENZYMATIC CRUDE ON MAGNETIC PARTICLES (Machine-translation by Google Translate, not legally binding) |
| WO2019002658A3 (en) * | 2017-06-28 | 2019-02-14 | Consejo Superior De Investigaciones Científicas (Csic) | Synthesis of biodiesel catalysed by an enzymatic crude immobilised on magnetic particles |
| CN108753761A (en) * | 2018-05-30 | 2018-11-06 | 北京理工大学珠海学院 | A kind of preparation method of amination modified composite magnetic nano-carrier for lipase immobilization |
| CN111041014A (en) * | 2019-12-31 | 2020-04-21 | 浙江工业大学 | Magnetic immobilized lipase and application thereof in resolution of 1-methyl-3-amphetamine |
| CN111041014B (en) * | 2019-12-31 | 2022-03-18 | 浙江工业大学 | Magnetic immobilized lipase and application thereof in resolution of 1-methyl-3-amphetamine |
| CN112680436A (en) * | 2020-12-23 | 2021-04-20 | 陕西科技大学 | Immobilized enzyme for extracting anthocyanin from black rice and preparation and use methods thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106119308A (en) | A kind of method that magnetic carrier immobilized-lipase prepares biodiesel | |
| Du et al. | Enzymatic synthesis of glycerol carbonate using a lipase immobilized on magnetic organosilica nanoflowers as a catalyst | |
| Aghaei et al. | Covalent immobilization of lipase from Candida rugosa on epoxy-activated cloisite 30B as a new heterofunctional carrier and its application in the synthesis of banana flavor and production of biodiesel | |
| Xie et al. | Magnetic solid catalysts for sustainable and cleaner biodiesel production: A comprehensive review | |
| Li et al. | Enhancing enzyme activity and enantioselectivity of Burkholderia cepacia lipase via immobilization on melamine-glutaraldehyde dendrimer modified magnetic nanoparticles | |
| Orçaire et al. | Application of silica aerogel encapsulated lipases in the synthesis of biodiesel by transesterification reactions | |
| Xie et al. | Immobilized lipase on Fe3O4 nanoparticles as biocatalyst for biodiesel production | |
| Li et al. | Synthesis of wax esters by lipase-catalyzed esterification with immobilized lipase from Candida sp. 99–125 | |
| CN104694526B (en) | Catalytic esterification and the Sn-1,3 selectivity immobilized lipase of transesterification and preparation method thereof | |
| Zeng et al. | Characterization of the lipase immobilized on Mg–Al hydrotalcite for biodiesel | |
| CN101412989B (en) | Method for synthesizing biodiesel by two-step catalysis of lipid via Yarrowia lipolytica lipase, and biodiesel obtained thereby | |
| Narwal et al. | Production and characterization of biodiesel using nonedible castor oil by immobilized lipase from Bacillus aerius | |
| Cao et al. | One-step direct transesterification of wet yeast for biodiesel production catalyzed by magnetic nanoparticle-immobilized lipase | |
| Jia et al. | Selective production of ethyl levulinate from levulinic acid by lipase-immobilized mesoporous silica nanoflowers composite | |
| Kumar et al. | Biodiesel production from transesterification of Serratia sp. ISTD04 lipids using immobilised lipase on biocomposite materials of biomineralized products of carbon dioxide sequestrating bacterium | |
| de Souza et al. | Immobilization of lipase B from Candida antarctica on epoxy‐functionalized silica: characterization and improving biocatalytic parameters | |
| Bi et al. | Biosynthesis of oleyl oleate in solvent-free system by Candida rugosa Lipase (CRL) immobilized in macroporous resin with cross-linking of aldehyde-dextran | |
| Paroul et al. | Solvent-free geranyl oleate production by enzymatic esterification | |
| CN101182507A (en) | Carrier and preparation and applications of immobilization lipase | |
| CN104313010A (en) | Preparation method of magnetic response composite lipase and application in biodiesel synthesis | |
| Abdulmalek et al. | Recent developments of lipase immobilization technology and application of immobilized lipase mixtures for biodiesel production | |
| Parandi et al. | Optimization of microreactor-intensified transesterification reaction of sesame cake oil (sesame waste) for biodiesel production using magnetically immobilized lipase nano-biocatalyst | |
| Santana et al. | Continuous flow reactor based with an immobilized biocatalyst for the continuous enzymatic transesterification of crude coconut oil | |
| CN101717801A (en) | Method for preparing biodiesel by using corn core immobilized lipase | |
| CN101671663B (en) | Preparation method of corncob fixed lipase and product thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161116 |