CN110408466A - A kind of method of enzyme assisting ion liquid extraction microalgae grease - Google Patents
A kind of method of enzyme assisting ion liquid extraction microalgae grease Download PDFInfo
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- CN110408466A CN110408466A CN201910716314.4A CN201910716314A CN110408466A CN 110408466 A CN110408466 A CN 110408466A CN 201910716314 A CN201910716314 A CN 201910716314A CN 110408466 A CN110408466 A CN 110408466A
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- microalgae grease
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- ion liquid
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- 239000004519 grease Substances 0.000 title claims abstract description 55
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 31
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 31
- 238000000605 extraction Methods 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 21
- 239000007788 liquid Substances 0.000 title claims abstract description 19
- 239000002608 ionic liquid Substances 0.000 claims abstract description 37
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 32
- 108010029541 Laccase Proteins 0.000 claims abstract description 23
- 239000006228 supernatant Substances 0.000 claims abstract description 22
- 241000195493 Cryptophyta Species 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000001816 cooling Methods 0.000 claims abstract description 11
- 239000000047 product Substances 0.000 claims abstract description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 9
- 230000007071 enzymatic hydrolysis Effects 0.000 claims abstract description 9
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims abstract description 9
- 230000010355 oscillation Effects 0.000 claims abstract description 9
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 9
- 239000000741 silica gel Substances 0.000 claims abstract description 9
- 229960001866 silicon dioxide Drugs 0.000 claims abstract description 9
- 230000007062 hydrolysis Effects 0.000 claims abstract description 5
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 5
- 238000000120 microwave digestion Methods 0.000 claims abstract description 5
- 150000002500 ions Chemical class 0.000 claims description 16
- 239000002253 acid Substances 0.000 claims description 15
- MCTWTZJPVLRJOU-UHFFFAOYSA-N 1-methyl-1H-imidazole Chemical compound CN1C=CN=C1 MCTWTZJPVLRJOU-UHFFFAOYSA-N 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 229920002472 Starch Polymers 0.000 claims description 7
- 235000019698 starch Nutrition 0.000 claims description 7
- 239000008107 starch Substances 0.000 claims description 7
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 6
- 108091005804 Peptidases Proteins 0.000 claims description 4
- -1 hexafluorophosphate Chemical compound 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- BNGXYYYYKUGPPF-UHFFFAOYSA-M (3-methylphenyl)methyl-triphenylphosphanium;chloride Chemical compound [Cl-].CC1=CC=CC(C[P+](C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 BNGXYYYYKUGPPF-UHFFFAOYSA-M 0.000 claims description 3
- 108091005508 Acid proteases Proteins 0.000 claims description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 3
- 102000035195 Peptidases Human genes 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 3
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 3
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- 229940124530 sulfonamide Drugs 0.000 claims description 3
- IBZJNLWLRUHZIX-UHFFFAOYSA-N 1-ethyl-3-methyl-2h-imidazole Chemical compound CCN1CN(C)C=C1 IBZJNLWLRUHZIX-UHFFFAOYSA-N 0.000 claims description 2
- 108090000604 Hydrolases Proteins 0.000 claims 1
- 102000004157 Hydrolases Human genes 0.000 claims 1
- 230000003301 hydrolyzing effect Effects 0.000 claims 1
- 238000011084 recovery Methods 0.000 abstract description 12
- 239000003960 organic solvent Substances 0.000 abstract description 7
- 239000000126 substance Substances 0.000 abstract description 5
- 238000004821 distillation Methods 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 description 19
- 229920005610 lignin Polymers 0.000 description 12
- 239000000243 solution Substances 0.000 description 10
- 239000000284 extract Substances 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 210000002421 cell wall Anatomy 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 238000005303 weighing Methods 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 230000002085 persistent effect Effects 0.000 description 5
- 239000008346 aqueous phase Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 238000006911 enzymatic reaction Methods 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000005855 radiation Effects 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 238000000638 solvent extraction Methods 0.000 description 3
- YQUVCSBJEUQKSH-UHFFFAOYSA-N 3,4-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 150000001335 aliphatic alkanes Chemical class 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- JMSVCTWVEWCHDZ-UHFFFAOYSA-N syringic acid Chemical compound COC1=CC(C(O)=O)=CC(OC)=C1O JMSVCTWVEWCHDZ-UHFFFAOYSA-N 0.000 description 2
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 102000029749 Microtubule Human genes 0.000 description 1
- 108091022875 Microtubule Proteins 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 150000003851 azoles Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003225 biodiesel Substances 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004939 coking Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000017858 demethylation Effects 0.000 description 1
- 238000010520 demethylation reaction Methods 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 108010093305 exopolygalacturonase Proteins 0.000 description 1
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 description 1
- 229940114124 ferulic acid Drugs 0.000 description 1
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 description 1
- 235000001785 ferulic acid Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000000762 glandular Effects 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000155 melt Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 210000004688 microtubule Anatomy 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- YIBXWXOYFGZLRU-UHFFFAOYSA-N syringic aldehyde Natural products CC12CCC(C3(CCC(=O)C(C)(C)C3CC=3)C)C=3C1(C)CCC2C1COC(C)(C)C(O)C(O)C1 YIBXWXOYFGZLRU-UHFFFAOYSA-N 0.000 description 1
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/04—Pretreatment of vegetable raw material
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
- C11B1/108—Production of fats or fatty oils from raw materials by extracting after-treatment, e.g. of miscellae
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/008—Refining fats or fatty oils by filtration, e.g. including ultra filtration, dialysis
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
A kind of method of enzyme assisting ion liquid extraction microalgae grease disclosed by the invention, the following steps are included: hydrophobic ionic liquid is added in (1) algae powder, 100-120 DEG C of micro-wave digestion, after cooling, laccase, hydrolysis complex enzyme is added, 40-60 DEG C of enzymatic hydrolysis algae powder dissolves out microalgae grease in hydrophobic ionic liquid；(2) oscillation layering, negated water phase, and it is cooled to 20-40 DEG C, centrifuging and taking supernatant；(3) supernatant is filtered through silicagel column to get microalgae grease finished product, and broken wall is high-efficient, is extracted without organic solvent up to microalgae grease, and recovery rate is high, consume energy low, no organic substance residues without distillation, and Environmental security coefficient is high, and extraction process step is simple and easy.
Description
Technical field
The present invention relates to microalgae grease fields, and in particular to a kind of method of enzyme assisting ion liquid extraction microalgae grease.
Background technique
Ecological environment caused by increasingly in short supply and fossil energy burning with petroleum resources is worsening, and biodiesel is made
Extensive concern is obtained for a kind of environmentally friendly renewable energy.
In numerous biomass, microalgae is with biomass is big, growth cycle is short, easy culture, contains more lipid
The effect of the advantages that matter, can directly generate hydrogen or produce oil product using fast pyrogenation and Direct liquefaction technology, and there are also carbon emission reductions,
Most microalgaes are all from nourishing one's nature, and very high using photosynthetic efficiency, there are many metabolites kinds liquid of generation, photosynthesis
Be it is unique it is a kind of can be by CO2It is converted into the process of organic principle, converts hydrogen, high unsaturated alkane for inorganic matter using solar energy
The energy substances such as hydrocarbon, grease, large-scale culturing micro-algae, unit area makes 10-20 times that has material crop, and microalgae adapts to energy
Power is strong, can grow under various water body environments, such as salt-soda soil, ocean, and microalgae can be used for sewage treatment and carbon capture,
Therefore microalgae is one of the energy for most having open future now.
Microalgae grease extracts the main method using " solvent extraction " and " mechanical expression " at present.Since most of oil-containing is micro-
Frustule wall is thicker, and when conventional organic solvent extracts, intracellular grease is difficult to emigrated cell wall, and solvent extraction rendering is time-consuming
It is long, and need to consume a large amount of organic extractants, the difficulty of separation is increased, and extraction efficiency is low；In addition extracted with organic solvent
Microalgae after taking, need to distill out organic solvent can just obtain grease, and energy consumption is high, and organic solvent determine whether to completely remove it is dry
Only, " mechanical expression " equally exists that recovery rate is relatively low, the high problem of energy consumption.
Summary of the invention
To solve the above problems, the present invention provides a kind of method of enzyme assisting ion liquid extraction microalgae grease, broken wall effect
Rate is high, extracts without organic solvent up to microalgae grease, and recovery rate is high, consume energy low, no organic substance residues without distillation, environment peace
Overall coefficient is high, and extraction process step is simple and easy.
A kind of method that the technical solution that the present invention solves the problems, such as is to provide enzyme assisting ion liquid extraction microalgae grease,
Characterized by comprising the following steps:
(1) hydrophobic ionic liquid is added in algae powder, after cooling, laccase, hydrolysis complex enzyme is added in 100-120 DEG C of micro-wave digestion,
40-60 DEG C of enzymatic hydrolysis algae powder dissolves out microalgae grease in hydrophobic ionic liquid；
(2) oscillation layering, negated water phase, and it is cooled to 20-40 DEG C, centrifuging and taking supernatant；
(3) supernatant is filtered through silicagel column to get microalgae grease finished product.
Preferably, the fusing point of the hydrophobic ionic liquid is 40-60 DEG C.
Preferably, the hydrophobic ionic liquid be one or more of.
Preferably, the hydrolysis complex enzyme includes one of acid pectase, acid protease, acid starch enzyme or several
Kind.
Preferably, the step (1) further includes adjusting pH value as 3.5-4.5.
Preferably, the step (1) is that hydrophobic ionic liquid is added in algae powder, and 100-120 DEG C of micro-wave digestion is cooling
Afterwards, it sequentially adds laccase, after non-proteolytic enzyme is stirred to react, is added proteolytic enzyme, 40-60 DEG C of enzymatic hydrolysis algae powder, dissolution is micro-
Algae oil rouge is in hydrophobic ionic liquid.
Preferably, the step (2) further includes that oil-in-water type demulsifier is added.
Preferably, the oil-in-water type demulsifier includes one of sodium chloride, magnesium chloride, calcium chloride, aluminum nitrate or several
Kind.
Preferably, the centrifugal speed is 3000-4000r/min.
Preferably, the enzymolysis time is 6-8h.
In the present solution, ionic liquid needs the recovery rate of microwave radiation exaraction nonpolarity content to greatly improve, this is because
Anhydrous broken wall is carried out to algae powder using microwave, by the ion conduction mechanism of microwave, ionic liquid can very fast and effeciently
Energy needed for reaction can be converted by microwave radiation, the structure of ionic liquid determine assimilation effect that it penetrates microwave good fortune very
Good, ionic liquid can extract more greases compared with conventional solvent, and hydrophobic grouping can shape in the solution in ionic liquid
At hydrophobic environment, hydrophobic channel can be formed in cell wall by hydrophobic grouping, facilitate the diffusion of nonpolar content, nonpolarity
Content further includes apolar substance not soluble in water other than microalgae grease.
However the process of microwave frequency measurment be frequency electromagnetic waves penetrate spe medium reach material inside microtubule fasolculus and glandular cell
Process, due to absorbing microwave energy, the temperature of cell interior will be risen rapidly, and heat is belonged to for the lignin in cell wall
It degrades, a large amount of free radical can be generated in lignin thermal degradation, these free radicals may polymerize generation coking again at high temperature
Phenomenon, it is therefore desirable to further be degraded to cell wall, and secondary dissolution microalgae grease, in addition cell obtained by microwave treatment
There can be a large amount of cell fragment, need to carry out enzymolysis processing to it；
Laccase plays oxidation, demethylation, polymerization and depolymerisation in lignin degradation, and lignin degradation generates syringic acid, perfume (or spice)
Oxalic acid, ferulic acid, protocatechuic acid etc. can also be used as laccase substrates and utilized, but laccase directly acts on the effect of lignin
Unobvious, this is because lignin is not soluble in water, therefore laccase and lignin, again in two reaction phases, the two contact area is very
Small, ionic liquid is the good solvent of lignin, and laccase acts on lignin in ionic liquid, can increase laccase with
The surface area of lignin contact, improves the function and effect of laccase, after laccase digests lignin, the enzymatic hydrolysis of final laccase it is unique
Product is water, can dissolve the product of complex enzyme zymohydrolysis cell, and protease hydrolyzed product and pectinase enzymatic hydrolysis product are that small molecule can
Molten object is separated by oscillation, the water phase of dissolution small molecule enzymolysis product and the nonaqueous phase comprising microalgae grease is obtained, wherein non-aqueous
The solvent of phase is hydrophobic ionic liquid, then by cooling, hydrophobic ionic liquid is condensed into solid, and microalgae grease at this time
Microalgae grease crude product can be obtained by filtering in liquid not yet；
In the present solution, the fusing point of hydrophobic ionic liquid needs within the scope of 30-70 DEG C, i.e. the optimum temperature of enzymatic reaction, temperature
Du Taigao then influences enzymatic reaction, and temperature is too low, solid is solidified as in enzymatic reaction, and the freezing point of grease is mostly also low
In 20 DEG C, microalgae DHA grease is even up to -44 DEG C, and when controlling temperature at 20-30 DEG C, hydrophobic ionic liquid is condensed into solid
Body, and microalgae grease is also liquid condition at this time, such hydrophobic ionic liquid include 1,3- methylimidazole hexafluorophosphate,
1,3- methylimidazole trifluoro sulfonamide, 1- ethyl -3- methyl-imidazoles hexafluorophosphate, 1- ethyl -3- dimethyl miaow
Azoles bis-trifluoromethylsulfoandimide salt, 1,3- methylimidazole hexafluorophosphate, the symmetry of these substance cationic structurals is low, from
It interacts between son weak, cation charge distribution is uniform, and anion volume is big, and fusing point is within the scope of 30-70 DEG C of enzyme activity.
It since enzymatic hydrolysis generates water, can have a negative impact to the extraction yield of grease, therefore demulsifier is added, electrolyte demulsification
Agent is reduced phase boundary potential and is changed the oleophylic water balance of emulsifier and worked by the interfacial electric double layer of compression emulsion drop.
This programme has the beneficial effect that
(1) using low melting point hydrophobic ionic liquid extract microalgae grease, using microalgae grease freezing point (fusing point) and dredge
The freezing point (fusing point) of aqueous ionic liquid is different, and microalgae grease is separated, without steaming again after adding organic solvent extraction
It evaporates and removes, consume energy low, organic solvent-free residual, Environmental security coefficient height；
(2) ionic liquid can increase the surface area that laccase is contacted with lignin, have preferable hydrolysis result to improve cell wall
The function and effect of laccase, and the enzymolysis product of laccase is water, dissolves the small molecule enzymolysis product of other complex enzymes, and and hydrophobicity
Ionic liquid layering achievees the purpose that extract microalgae grease；
(3) energetic ion molecule sizes needed for ionic liquid very fast and effeciently can convert reaction for microwave radiation
Conventional solvent can extract more greases, and hydrophobic grouping can form hydrophobic environment in the solution in ionic liquid, by dredging
Water base group can form hydrophobic channel in cell wall, facilitate the diffusion of nonpolar content.
Specific embodiment
In order to make content of the present invention easily facilitate understanding, With reference to embodiment to of the present invention
Technical solution is described further, but the present invention is not limited only to this.
Embodiment 1
100mg algae powder be added 5ml hydrophobic ionic liquid 1,3- methylimidazole hexafluorophosphate ,-rise be put into counteracting tank,
It (need to symmetrically be set) in merging rotor, open rear microwave reactor, temperature setting is 120 DEG C, heating power 800W, one timing of reaction
Between after, it is cooling to system, melt counteracting tank, 10U be added and purifies Tversicolor sdu-4 laccase, 60 DEG C of oscillating reactions 2h,
PH value is adjusted to 3.5, is added 10U acid pectase, 10U acid starch enzyme, 60 DEG C of oscillating reactions 3h, persistent oscillation extraction takes
Non-aqueous solution, and 30 DEG C are cooled to, 3000r/min is centrifuged 15min, takes supernatant liquor, and solid sodium chloride, 3000r/ is added
Min is centrifuged 15min, takes supernatant liquor, repeats this step three times, merges supernatant liquor, is filtered with silicagel column up to microalgae grease,
With its quality of electronic balance weighing, be computed microalgae grease recovery rate is 87%.
Embodiment 2
100mg algae powder be added 5ml hydrophobic ionic liquid 1,3- methylimidazole trifluoro sulfonamide ,-rise be put into resolution
In tank, being placed in rotor (need to symmetrically be set), open rear microwave reactor, and temperature setting is 100 DEG C, heating power 800W, reaction
After a certain period of time, cooling to system, counteracting tank is melted, 10U is added and purifies Tversicolor sdu-4 laccase, it is acid that 10U is added
Pectase, 10U acid starch enzyme, 60 DEG C of oscillating reactions 4h, persistent oscillation extraction, negated aqueous phase solution, and 30 DEG C are cooled to,
3000r/min is centrifuged 15min, takes supernatant liquor, and solid sodium chloride is added, and 3000r/min is centrifuged 15min, takes supernatant liquor, weight
This multiple step three times, merges supernatant liquor, is filtered with silicagel column up to microalgae grease, with its quality of electronic balance weighing, through counting
Calculate microalgae grease recovery rate be 82%.
Embodiment 3
100mg algae powder be added 5ml hydrophobic ionic liquid 1- ethyl -3- methyl-imidazoles hexafluorophosphate,-rise be put into counteracting tank
In, being placed in rotor (need to symmetrically set), open rear microwave reactor, and temperature setting is 120 DEG C, heating power 800W, reaction one
It is cooling to system after fixing time, counteracting tank is melted, 10U is added and purifies Tversicolor sdu-4 laccase, 70 DEG C of oscillating reactions
2h adjusts pH value to 4.5,10U acid pectase, 10U acid starch enzyme, 70 DEG C of oscillating reactions 4h, persistent oscillation extraction is added
It taking, negated aqueous phase solution, and is cooled to 20 DEG C, 3500r/min is centrifuged 15min, takes supernatant liquor, aluminum nitrate solid is added,
3000r/min is centrifuged 15min, takes supernatant liquor, repeats this step three times, merges supernatant liquor, is filtered with silicagel column up to micro-
Algae oil rouge, with its quality of electronic balance weighing, be computed microalgae grease recovery rate is 79%.
Embodiment 4
5ml hydrophobic ionic liquid 1- ethyl -3- methylimidazole bis-trifluoromethylsulfoandimide salt is added in 100mg algae powder,-rise
It is put into counteracting tank, being placed in rotor (need to symmetrically set), open rear microwave reactor, and temperature setting is 120 DEG C, and heating power is
800W, reaction is after a certain period of time, cooling to system, melts counteracting tank, and 10U is added and purifies Tversicolor sdu-4 laccase, and 30
DEG C oscillating reactions 2h adjusts pH value to 4.5,10U acid pectase, 10U acid starch enzyme is added, 30 DEG C of oscillating reactions 4h are held
Persistent oscillation extraction, negated aqueous phase solution, and 20 DEG C are cooled to, 3000r/min is centrifuged 15min, takes supernatant liquor, and calcium chloride is added
Solid, 4000r/min are centrifuged 15min, take supernatant liquor, repeat this step three times, merge supernatant liquor, are with silicagel column filtering
Microalgae grease, with its quality of electronic balance weighing, be computed microalgae grease recovery rate is 81%.
Embodiment 5
100mg algae powder be added 5ml hydrophobic ionic liquid 1,3- methylimidazole hexafluorophosphate ,-rise be put into counteracting tank,
It (need to symmetrically be set) in merging rotor, open rear microwave reactor, temperature setting is 120 DEG C, heating power 800W, one timing of reaction
Between after, it is cooling to system, melt counteracting tank, 10U be added and purifies Tversicolor sdu-4 laccase, 40 DEG C of oscillating reactions 2h are adjusted
PH value is saved to 4.5,10U acid pectase is added, 10U acid starch enzyme, 40 DEG C of oscillating reactions 4h, persistent oscillation extracts, negated
Aqueous phase solution, and 20 DEG C are cooled to, 3000r/min is centrifuged 15min, takes supernatant liquor, and magnesium chloride solids, 3000r/min is added
It is centrifuged 15min, takes supernatant liquor, repeats this step three times, merges supernatant liquor, is filtered with silicagel column up to microalgae grease, is used
Its quality of electronic balance weighing, be computed microalgae grease recovery rate be 84%.
Embodiment 6
100mg algae powder is dissolved in 100ml phosphate buffer, and pH is adjusted to 4.5,10U is added and purifies Tversicolor sdu-4
Laccase, 10U acid pectase, 40 DEG C of oscillating reactions 6h add 10U acid protease, 40 DEG C of oscillating reactions 6h, be added just oneself
Alkane extracting and demixing merges supernatant liquor, is filtered with silicagel column and is computed up to microalgae grease with its quality of electronic balance weighing
Obtaining microalgae grease recovery rate is 83%.
It is found that the recovery rate using this programme microalgae grease is close with enzymatic hydrolysis extraction recovery rate used in comparative example 5,
Illustrate that this programme is feasible.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (10)
1. a kind of method of enzyme assisting ion liquid extraction microalgae grease, which comprises the following steps:
(1) hydrophobic ionic liquid is added in algae powder, after cooling, laccase, hydrolysis complex enzyme is added in 100-120 DEG C of micro-wave digestion,
30-70 DEG C of enzymatic hydrolysis algae powder dissolves out microalgae grease in hydrophobic ionic liquid；
(2) oscillation layering, negated water phase, and it is cooled to 20-30 DEG C, centrifuging and taking supernatant；
(3) supernatant is filtered through silicagel column to get microalgae grease finished product.
2. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that described
The fusing point of hydrophobic ionic liquid is 30-70 DEG C.
3. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that described
Hydrophobic ionic liquid is 1,3- methylimidazole hexafluorophosphate, 1,3- methylimidazole trifluoro sulfonamide, 1- second
Base -3- methyl-imidazoles hexafluorophosphate, 1- ethyl -3- methylimidazole bis-trifluoromethylsulfoandimide salt, 1,3- methylimidazole
One or more of hexafluorophosphate.
4. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that described
Hydrolyzing complex enzyme includes one or more of acid pectase, acid protease, acid starch enzyme.
5. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that described
Step (1) further includes adjusting pH value as 3.5-4.5.
6. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that described
Step (1) is that hydrophobic ionic liquid is added in algae powder, and 100-120 DEG C of micro-wave digestion sequentially adds laccase, non-egg after cooling
After white hydrolase is stirred to react, proteolytic enzyme is added, 30-70 DEG C of enzymatic hydrolysis algae powder dissolves out microalgae grease in Hydrophobic Ionic liquid
Body.
7. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that described
Step (2) further includes that oil-in-water type demulsifier is added.
8. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 7, which is characterized in that described
Oil-in-water type demulsifier includes one or more of sodium chloride, magnesium chloride, calcium chloride, aluminum nitrate.
9. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that described
Centrifugal speed is 3000-4000r/min.
10. a kind of method of enzyme assisting ion liquid extraction microalgae grease according to claim 1, which is characterized in that institute
Stating enzymolysis time is 6-8h.
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| CN111690462A (en) * | 2020-06-09 | 2020-09-22 | 厦门汇盛生物有限公司 | Method for demulsifying and extracting oil from oil-containing algae or fungus cell wall-broken liquid |
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