CN109569728A - A kind of biology base metal organic frame assembles esterase catalyzed dose, preparation method and its application in biodiesel synthesis - Google Patents
A kind of biology base metal organic frame assembles esterase catalyzed dose, preparation method and its application in biodiesel synthesis Download PDFInfo
- Publication number
- CN109569728A CN109569728A CN201811550027.2A CN201811550027A CN109569728A CN 109569728 A CN109569728 A CN 109569728A CN 201811550027 A CN201811550027 A CN 201811550027A CN 109569728 A CN109569728 A CN 109569728A
- Authority
- CN
- China
- Prior art keywords
- qlm
- esterase
- mof
- biology base
- base metal
- 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
- 108090000371 Esterases Proteins 0.000 title claims abstract description 42
- 239000003225 biodiesel Substances 0.000 title claims abstract description 32
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 26
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 26
- 239000010953 base metal Substances 0.000 title claims abstract description 15
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 72
- 235000019486 Sunflower oil Nutrition 0.000 claims abstract description 22
- 239000002600 sunflower oil Substances 0.000 claims abstract description 22
- 239000003054 catalyst Substances 0.000 claims abstract description 19
- 239000012621 metal-organic framework Substances 0.000 claims description 51
- 238000006243 chemical reaction Methods 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 20
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 14
- 229930024421 Adenine Natural products 0.000 claims description 14
- 229960000643 adenine Drugs 0.000 claims description 14
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 239000012918 MOF catalyst Substances 0.000 claims description 7
- 230000001376 precipitating effect Effects 0.000 claims description 7
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000004246 zinc acetate Substances 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 5
- 238000002390 rotary evaporation Methods 0.000 claims description 5
- 239000006185 dispersion Substances 0.000 claims description 4
- 239000013110 organic ligand Substances 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 239000000758 substrate Substances 0.000 claims description 2
- 239000012153 distilled water Substances 0.000 claims 1
- 239000002244 precipitate Substances 0.000 claims 1
- 238000010025 steaming Methods 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 abstract description 23
- 102000004190 Enzymes Human genes 0.000 abstract description 23
- 238000005809 transesterification reaction Methods 0.000 abstract description 13
- 230000003197 catalytic effect Effects 0.000 abstract description 11
- 239000000463 material Substances 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 9
- 238000004064 recycling Methods 0.000 abstract description 6
- 230000007062 hydrolysis Effects 0.000 abstract description 5
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 5
- HGLRAOXHGPCZQH-UHFFFAOYSA-N C(CCCCCCC)(=O)O.[N+](=O)([O-])C1=CC=C(C(=O)C2=CC=CC=C2)C=C1 Chemical compound C(CCCCCCC)(=O)O.[N+](=O)([O-])C1=CC=C(C(=O)C2=CC=CC=C2)C=C1 HGLRAOXHGPCZQH-UHFFFAOYSA-N 0.000 abstract description 4
- 239000003921 oil Substances 0.000 abstract description 3
- 235000019198 oils Nutrition 0.000 abstract description 3
- 150000002148 esters Chemical class 0.000 abstract description 2
- 102000018120 Recombinases Human genes 0.000 abstract 1
- 108010091086 Recombinases Proteins 0.000 abstract 1
- 230000008569 process Effects 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 230000002255 enzymatic effect Effects 0.000 description 8
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 239000002283 diesel fuel Substances 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 230000003592 biomimetic effect Effects 0.000 description 5
- 230000033558 biomineral tissue development Effects 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- 239000002585 base Substances 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000007613 environmental effect Effects 0.000 description 4
- -1 carbomethoxy Chemical group 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 235000019387 fatty acid methyl ester Nutrition 0.000 description 3
- 150000004665 fatty acids Chemical class 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000005311 nuclear magnetism Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 238000001338 self-assembly Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 230000004580 weight loss Effects 0.000 description 2
- ZYMCBJWUWHHVRX-UHFFFAOYSA-N (4-nitrophenyl)-phenylmethanone Chemical compound C1=CC([N+](=O)[O-])=CC=C1C(=O)C1=CC=CC=C1 ZYMCBJWUWHHVRX-UHFFFAOYSA-N 0.000 description 1
- 241000588810 Alcaligenes sp. Species 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000010775 animal oil Substances 0.000 description 1
- 238000005815 base catalysis Methods 0.000 description 1
- 230000002210 biocatalytic effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 239000002385 cottonseed oil Substances 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000010931 ester hydrolysis Methods 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000021323 fish oil Nutrition 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000010408 sweeping Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/16—Catalysts comprising hydrides, coordination complexes or organic compounds containing coordination complexes
- B01J31/1691—Coordination polymers, e.g. metal-organic frameworks [MOF]
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/16—Catalysts comprising hydrides, coordination complexes or organic compounds containing coordination complexes
- B01J31/22—Organic complexes
- B01J31/2204—Organic complexes the ligands containing oxygen or sulfur as complexing atoms
- B01J31/2208—Oxygen, e.g. acetylacetonates
- B01J31/2213—At least two complexing oxygen atoms present in an at least bidentate or bridging ligand
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J35/00—Catalysts, in general, characterised by their form or physical properties
- B01J35/60—Catalysts, in general, characterised by their form or physical properties characterised by their surface properties or porosity
- B01J35/61—Surface area
-
- C—CHEMISTRY; METALLURGY
- C10—PETROLEUM, GAS OR COKE INDUSTRIES; TECHNICAL GASES CONTAINING CARBON MONOXIDE; FUELS; LUBRICANTS; PEAT
- C10L—FUELS NOT OTHERWISE PROVIDED FOR; NATURAL GAS; SYNTHETIC NATURAL GAS OBTAINED BY PROCESSES NOT COVERED BY SUBCLASSES C10G, C10K; LIQUEFIED PETROLEUM GAS; ADDING MATERIALS TO FUELS OR FIRES TO REDUCE SMOKE OR UNDESIRABLE DEPOSITS OR TO FACILITATE SOOT REMOVAL; FIRELIGHTERS
- C10L1/00—Liquid carbonaceous fuels
- C10L1/02—Liquid carbonaceous fuels essentially based on components consisting of carbon, hydrogen, and oxygen only
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C3/00—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom
- C11C3/04—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom by esterification of fats or fatty oils
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C3/00—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom
- C11C3/04—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom by esterification of fats or fatty oils
- C11C3/10—Ester interchange
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2531/00—Additional information regarding catalytic systems classified in B01J31/00
- B01J2531/50—Complexes comprising metals of Group V (VA or VB) as the central metal
- B01J2531/54—Bismuth
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Materials Engineering (AREA)
- General Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Enzymes And Modification Thereof (AREA)
- Fats And Perfumes (AREA)
Abstract
A kind of biology base metal organic frame assembles esterase catalyzed dose, preparation method and its application in biodiesel synthesis, belongs to field of biotechnology.The present invention is immobilized among biology base MOF material by thermophilic esterase QLM, finds that the immobilized esterase among MOF still has good catalytic activity by Activity determination, can successfully be catalyzed the hydrolysis of esters p-NPC (4- nitrobenzophenone caprylate)；Using above-mentioned esterase-MOF assembly as catalyst, it is catalyzed the transesterification reaction of sunflower oil and methanol, successfully synthesizes biodiesel of good performance.After thermophilic esterase QLM enzyme is supported into biology base MOF, there is better thermal stability and pH tolerance；Using the immobilised enzymes as catalyst, the synthesis of biodiesel can be successfully realized, had under high oil/methanol ratio compared with the higher catalytic efficiency of resolvase, while there is good operational stability and recycling ability.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of biological Base Metal based on the building of biomimetic mineralization technology has
Machine frame assemble esterase catalyzed dose, preparation method and its biodiesel synthesis in application.
Background technique
Biodiesel refer to vegetable oil (such as sunflower oil, rapeseed oil, soybean oil, peanut oil, corn oil, cottonseed oil),
Animal oil (such as fish oil, lard, butter, sheep oil), waste grease or microbial oil and alcohols are by transesterification and forming
Fatty acid methyl ester or ethyl ester.Biodiesel is typical " green energy resource ", have good environmental protection, startup performance of engine it is good,
Fuel performance is good, raw material sources are extensive, the characteristics such as renewable.Biodiesel is greatly developed to sustainable economic development, propulsion energy
Source substitution mitigates environmental pressure etc. with important strategic importance.Traditional biodiesel mainly by the strategy of base catalysis come
It is prepared, compared with traditional method for synthesizing biologic diesel oil, the method for esterase catalyzed biodiesel synthesis has synthesis process
Simply, the advantages that bubble that energy consumption is few, generates is few, has broad application prospects.
Esterase (Esterase, EC 3.1.1.1) is a kind of serine hydrolase class for being distributed widely in tissue and organ,
Can hydrolysis it is many containing ester bond, thioester bond, amido bond endogenous and exogenous material.Its major function is to participate in lipid generation
Thank, signal transduction and maintain biofilm structure integrality, the crowds such as transesterification, esterification and transesterification can also be completed in organic phase
More reactions.In recent years, people's isolated thermophilic esterase from thermophilic microorganism, since its stability is high, preparation cost is low
The advantages that and be concerned, in modern technical field of enzyme engineering have important application value.It is thermophilic employed in this patent
Hot esterase QLM has good thermal stability and catalytic activity in alcaligenes Alcaligenes sp., most suitable
Temperature is at 65~70 DEG C.
Although esterase have the advantages that it is above-mentioned so many, it in industrial biocatalytic application still face it is very big
Challenge, such as low stability under industrial process conditions and recycling rate of waterused etc..Enzyme immobilization technology is to solve above-mentioned bottleneck
An effective way, the good catalysis activity of enzyme molecule, stability can be assigned, to the highly resistant of environment and repeat benefit
Use ability.In the material of enzyme immobilization, by metal ion and organic ligand by being coordinated the crystalline material-- metal formed
Organic frame (Metal-organic framework, MOF), since it has composition and diversity of pattern, pore size
The advantages that controllability, excellent specific surface area etc. and be widely used.In recent years, it is fixed about by matrix progress enzyme of MOF
The report of change is concentrated mainly on two aspects: enzyme being adsorbed among the aperture MOF by way of physical absorption；By covalently connecting
It connects mode enzyme is coupled on MOF.But both modes all have the defect of oneself, such as physical adsorption way respectively, enzyme holds
Easily fall off from MOF skeleton, and covalent attachment then will affect the catalytic activity of enzyme.Compared with above two strategy, bionical mine
Change technology is a kind of novel enzyme immobilization strategy based on MOF matrix newly developed in recent years, is imitated such as molluscan
The forming processes such as shell, the gum of mammal, bone, the self assembly of first synthetic biological molecule (protein, lipid, nucleic acid etc.)
Body forms organic/inorganic composite then under the action of self-assembly template.It is bionical compared with traditional immobilization strategy
Mineralising immobilization strategy can be immobilized into MOF molecule by biggish biomolecule, and to the conformation of enzyme molecule and active shadow
Sound is smaller, with good stability.
Summary of the invention
The present invention is immobilized among biology base MOF material by thermophilic esterase QLM by the method for biomimetic mineralization immobilization,
It finds that the immobilized esterase among MOF still has good catalytic activity by Activity determination, can successfully be catalyzed esters p-
The hydrolysis of NPC (4- nitrobenzophenone caprylate)；Using above-mentioned esterase-MOF assembly as catalyst, it is catalyzed sunflower oil and methanol
Transesterification reaction successfully synthesizes biodiesel of good performance.
A kind of biology base metal organic frame of the present invention assembles esterase catalyzed dose of preparation method, and feature exists
In: organic ligand molecule adenine aqueous solution (80~100 DEG C) is poured into water-soluble dissolved with thermophilic esterase QLM and zinc acetate
In liquid, react 24~48 hours at room temperature；After completion of the reaction, reaction system is centrifuged 3~8min through 5000~10000r/min, receives
Collection precipitating, precipitating obtain biology base metal organic frame assembling esterase of the present invention through distilling water washing, after freeze-drying and urge
Agent (QLM@Bio-MOF).
In adenine aqueous solution, the concentration of adenine is 50~150mg/mL；The quality of thermophilic esterase QLM and adenine is used
Amount ratio is 0.02~0.2:1, and the quality usage ratio of zinc acetate and adenine is 0.2~0.6:1.
Further, using biology base metal organic frame assembling esterase as catalyst, using sunflower oil as substrate, in methanol
Transesterification reaction is carried out in system, 40~70 DEG C are reacted 2~8 hours, obtain biodiesel by extraction and rotary evaporation.Specifically
It is:
1. QLM@Bio-MOF 80~120mg of catalyst is added, ultrasonic disperse is in 20~40mL sunflower oil.
2. methanol is added into above-mentioned dispersion liquid, the volume ratio of sunflower oil and methanol is 2~8:1, and uses 1M NaOH
PH=8~10 for adjusting reaction solution, are then vigorously stirred under (600~800rpm) at 40~70 DEG C and react 2~8 hours；
3. being filtered to remove QLM@Bio-MOF catalyst, using separatory funnel separation supernatant liquor and rotary evaporation is removed
Unreacted methanol in layer clear liquid, obtains product biodiesel.
Biology base metal organic frame prepared by the present invention assembles esterase catalyzed dose (QLM@Bio-MOF) with following excellent
Point: environmentally protective, environmental pollution caused by catalytic process is less；Good biocompatibility, if applied in vivo, Bu Huizao
It is reacted at biocompatible；Combination between ligand is stronger, and the more general MOF material of stability is stronger.
The present invention assembles esterase (QLM Bio- using sunflower oil as raw material, with above-mentioned biology base metal organic frame
MOF it is) catalyst, transesterification reaction is catalyzed in methanol system, carries out the synthesis of biodiesel.It is prepared with traditional biodiesel
Process is compared, which has the advantage that synthesis process green non-pollution；Synthesis process is smaller on enzymatic activity influence, is easy
Realize the cycling and reutilization of catalyst；The biodiesel of synthesis has excellent environmental protection characteristic, safe and reliable.
To sum up, biocompatibility adenine of the present invention is the building that organic ligand carries out MOF system, passes through 1 He of embodiment
Biomimetic mineralization immobilization technology in embodiment 2 carries out the immobilization of thermophilic esterase, and is to urge with above-mentioned immobilization thermophilic esterase
Agent carries out transesterification reaction of the sunflower oil in methanol system, is successfully prepared biodiesel.In the catalyst system, urge
Agent can repeated recycling utilize, cost is greatly reduced, and simultaneously synthesizing process is easy, and safety is pollution-free.The present invention is expected to become
A kind of new technique means of biodiesel synthesis to sustainable economic development, promote the tools such as energy substitution, mitigation environmental pressure
There is important meaning.There is better catalytic activity and steady thermophilic esterase catalyzed dose of biology base MOF- constructed by present patent application
It is qualitative, it is expected to become a kind of good biological catalyst in the preparation of future biological diesel oil.
Detailed description of the invention
Fig. 1: the stereoscan photograph for the QLM@Bio-MOF that embodiment 1 synthesizes.
Fig. 2: the scanning electron microscope result for the QLM@Bio-MOF that embodiment 2 synthesizes.
The thermogravimetric curve of Fig. 3: QLM@Bio-MOF and me-Bio-MOF.Abscissa is temperature in figure, and ordinate is material
Weight.
Fig. 4: influence curve of the temperature to QLM@Bio-MOF and QLM enzymatic activity.Abscissa is temperature in figure, and ordinate is
Relative activity
Fig. 5: pH to the influence curve of QLM@Bio-MOF and QLM enzymatic activity.Abscissa is pH value, ordinate in figure
For relative activity.
Fig. 6: being the nuclear-magnetism figure of catalyst biodiesel synthesis using QLM@Bio-MOF constructed by the present invention.It is horizontal in figure
Coordinate is chemical shift.
Fig. 7: being the conversion ratio column diagram of catalyst biodiesel synthesis using QLM@Bio-MOF constructed by the present invention.
Abscissa is the volume ratio of sunflower oil and methanol, the conversion ratio of ordinate sunflower oil in figure.
Fig. 8: QLM@Bio-MOF catalysis biological diesel oil synthesizes recycling ability column diagram.Abscissa is material in figure
Number is recycled, ordinate is the conversion ratio of sunflower oil.
Specific embodiment
Example given below is that the invention will be further described, more fully in order to those skilled in the art
Understand the present invention.But given embodiment should not be understood as limiting the scope of the invention, thus the technology of the profession
Personnel also should belong to the scope of the present invention according to the nonessential modifications and adaptations that foregoing invention content is made.
Embodiment 1
The adenine of 500mg is dissolved in the 5mL boiling water that temperature is 100 DEG C, then pours into the adenine solution rapidly
Dissolved in the 20mL of 10mg thermophilic esterase QLM and 200mg zinc acetate, 25 DEG C of aqueous solutions, 48h is reacted at room temperature.After completion of the reaction,
System is centrifuged 5min through 8000r/min, collects precipitating, three times through distillation water washing, the powder of freeze-drying is final product to precipitating
QLM@Bio-MOF, product quality 250mg.
By similar methods, preparation does not support the biology base MOF material (me-Bio-MOF) of thermophilic esterase, and difference is
Thermophilic esterase QLM is not added.
It is found from the scanning electron microscope of Fig. 1 synthetic product, using the shape of QLM@Bio-MOF catalyst synthesized by the above method
Looks are regular, and partial size is between 800nm~1.5 μm.
Embodiment 2
The adenine of 1000mg is dissolved in 100 DEG C of temperature of 10mL boiling water, then pours into the adenine solution rapidly
Dissolved with the 200mL of 100mg QLM and 400mg zinc acetate, among 25 DEG C of aqueous solutions.48h is reacted at room temperature.After completion of the reaction, body
System is centrifuged 5min through 8000r/min, collects precipitating, three times through distillation water washing, the powder of freeze-drying is final product QLM to precipitating
Bio-MOF, product quality 450mg.
By similar methods, preparation does not support the biology base MOF material (me-Bio-MOF) of thermophilic esterase, and difference is
Thermophilic esterase QLM is not added.
As a result, it has been found that, expand the QLM@of raw material input amount synthesis by equal proportion from the surface sweeping Electronic Speculum of Fig. 2 synthetic product
Bio-MOF regular appearance is uniform, and partial size is between 800nm~1.5 μm.The QLM@Bio- obtained compared to small lot synthetic method
MOF is not substantially change on pattern and partial size.
Embodiment 3
The QLM Bio-MOF and me-Bio-MOF that embodiment 1 is synthesized carry out thermal weight loss experiment after sufficiently drying, and use TA
Q500 detecting instrument is heated up in nitrogen environment with the rate of 10 DEG C/min, and thermogravimetric curve obtained is as shown in Figure 2.
From Fig. 3 it can be found that QLM@Bio-MOF has apparent weightlessness at 200~500 DEG C, the weightlessness than me-Bio-MOF
Measure it is high by 20% or so, height come out this part weight loss represent weight ratio shared by enzyme.Therefore, it according to thermogravimetric curve, calculates
The loading of QLM is 20% or so in QLM@Bio-MOF.
Embodiment 4
It is reacted using the hydrolysis of p-NPC (4- nitrobenzophenone caprylate) as mode, evaluation temperature and pH are to QLM@
The influence of the catalytic and stability of Bio-MOF.
Influence of the temperature to enzyme activity is detected, the acetonitrile solution (10mM) of 40 μ L p-NPC is added to the phosphate of 920 μ L
In buffer (50mM, pH 8.0), be then incubated for 1min under different temperatures (30,40,50,60,70,80,90 DEG C), finally plus
Enter the QLM@Bio-MOF starting reaction of the QLM enzyme solutions (1mg/mL) or identical equivalent of 40 μ L, reaction time 1min.It is right
Enzyme is not added according to product, remaining composition is consistent with the above, and control group is used to deduct the background of ester hydrolysis reaction.Then use 1cm mouthfuls
For the cuvette of diameter in the generation of 405nm detection product p- nitrobenzophenone, detecting instrument is UV-7500 Ultraviolet Detector.Definition
The enzyme amount that 1min discharges 1 μM of p-nitrophenyl is an enzyme activity unit.
Influence of the pH value to enzyme activity is detected, we equally select the hydrolysis of p-NPC (4- nitrobenzophenone caprylate)
It is reacted as mode.The acetonitrile solution (10mM) of 20 μ L p-NPC be added to 960 μ L different pH value (6.0,6.5,7.0,7.5,
8.0,8.5,9.0) in buffer, then in 60 DEG C of incubation 1min, be eventually adding 20 μ L QLM enzyme solutions (1mg/mL) or
The QLM@Bio-MOF of identical equivalent originates reaction, reaction time 1min.Remaining operating process and above-mentioned temperature are to enzyme activity shadow
Loud detection method is consistent.
From Fig. 4 it can be found that the optimum temperature of QLM@Bio-MOF is shifted to the right to 65 DEG C from 60 DEG C, after illustrating biomimetic mineralization
Esterase has better enzymatic activity at a higher temperature, compared to free enzyme.Even more noteworthy, in high temperature (80-90
DEG C) under the conditions of, the enzymatic activity of QLM@Bio-MOF is substantially better than free QLM, and the esterase after illustrating mineralising has better resistance to height
The performance of temperature.
It is found from Fig. 5, under conditions of slant acidity (pH≤6.5) and meta-alkalescence (pH >=7.5), QLM@Bio-MOF's
Catalytic activity is better than free QLM, and the esterase after illustrating mineralising has the stronger ability for resisting acid or alkali environment.
Embodiment 5
It is that catalyst (constructs two in parallel using QLM@Bio-MOF or free QLM using sunflower oil as raw material
A reaction system, one is catalyst using QLM@Bio-MOF, and one is catalyst using QLM), it is catalyzed transesterification reaction, synthesis
The specific operation process of biodiesel is as follows:
1. the QLM of QLM@Bio-MOF catalyst 100mg or identical equivalent is added, ultrasonic disperse is in 30mL sunflower oil
In.
2. methanol is added into above-mentioned dispersion liquid, the volume ratio of sunflower oil and methanol is 2:1, and uses 1M NaOH tune
The pH=8 for saving reaction solution, is then vigorously stirred under (800rpm) at 50 DEG C and reacts 5h.
3. being filtered to remove QLM@Bio-MOF catalyst, using separatory funnel separation supernatant liquor and rotary evaporation is removed
Unreacted methanol in layer clear liquid, obtains product biodiesel.
4. product fatty acid methyl esters passes through1H NMR is detected, if (characteristic peak of carbomethoxy) goes out at 3.63ppm
Existing characteristic peak then proves that fatty acid and methanol realize transesterification successfully to biodiesel synthesis.By
Integrating peak areas at 3.63ppm and at methylene characteristic peak 2.30ppm, available conversion ratio.
Fig. 6 nuclear-magnetism testing result can be found that the carbomethoxy characteristic peak at 3.63, it was demonstrated that urging by QLM@Bio-MOF
Change, fatty acid and methanol realize transesterification to synthesize biodiesel.
Embodiment 6
It is that catalyst (constructs two in parallel using QLM@Bio-MOF or free QLM using sunflower oil as raw material
A reaction system, one is catalyst using QLM@Bio-MOF, and one is catalyst using QLM), it is catalyzed transesterification reaction, synthesis
The specific operation process of biodiesel is as follows:
1. QLM@Bio-MOF catalyst 100mg is added, and the QLM of identical equivalent, ultrasonic disperse are added according to loading
In 30mL sunflower oil.
2. methanol is added in above-mentioned dispersion liquid, the volume ratio of sunflower oil and methanol is 2:1,4:1,6:1 and 8:1, is used
1M NaOH adjusts the pH to 8.5 of reaction solution, then reacts 5h under the conditions of 50 DEG C, synthesis process needs be vigorously stirred (600~
800rpm)。
3. being filtered to remove QLM@Bio-MOF catalyst, using separatory funnel separation supernatant liquor and rotary evaporation is removed
Unreacted methanol in layer clear liquid, obtains product biodiesel.
4. product fatty acid methyl esters passes through1H NMR is detected, if (characteristic peak of carbomethoxy) goes out at 3.63ppm
Existing characteristic peak then proves that fatty acid and methanol realize transesterification successfully to biodiesel synthesis.By
Integrating peak areas at 3.63ppm and at methylene characteristic peak 2.30ppm, available conversion ratio.
Conversion ratio under different sunflower oils, methanol ratio is as shown in fig. 7, discovery increases with sunflower oil, methanol ratio
Add, conversion ratio gradually decreases.And QLM Bio-MOF is used to obtain as the diesel conversion of catalyst and using free enzymatic
Conversion ratio it is suitable, and under the conditions of 8:1, conversion ratio is slightly above the conversion ratio of free enzymatic.Illustrate using biomimetic mineralization
The QLM@Bio-MOF of technology synthesis can be very good to keep the enzymatic activity of esterase QLM, the energy with the synthesis of good catalytic diesel oil
Power.
Embodiment 7
After the synthesis of first diesel oil, QLM@Bio-MOF is collected by centrifugation, the conjunction of next group biodiesel is used for after freeze-drying
At, and so on, until recycling the 6th time.The conversion of each catalytic diesel oil synthesis is obtained according to the calculation method of embodiment 5
Rate.
It being found from Fig. 8, QLM Bio-MOF can still be kept above 50% conversion ratio after continuously recycling 6 times,
Illustrate that the material has and preferably recycle stability, meets the condition required for industrial application compared with high circulation utilization rate,
It is suitable for industrial production and application.
Claims (5)
1. a kind of biology base metal organic frame assembles esterase catalyzed dose of preparation method, it is characterised in that: by 80~100 DEG C
Organic ligand molecule adenine aqueous solution is poured into the aqueous solution dissolved with thermophilic esterase QLM and zinc acetate, is reacted at room temperature
24~48 hours；After completion of the reaction, reaction system is centrifuged 3~8min through 5000~10000r/min, collects precipitating, precipitates through steaming
Distilled water washing, i.e. obtaining biology base metal organic frame assembles esterase catalyzed dose of QLM Bio-MOF after freeze-drying.
2. a kind of biology base metal organic frame as described in claim 1 assembles esterase catalyzed dose of preparation method, feature
Be: the concentration of adenine is 50~150mg/mL, and the quality usage ratio of thermophilic esterase QLM and adenine is 0.02~0.2:
1, the quality usage ratio of zinc acetate and adenine is 0.2~0.6:1.
3. a kind of biology base metal organic frame assembles esterase catalyzed dose, it is characterised in that: be by of any of claims 1 or 2
Method is prepared.
4. biology base metal organic frame as claimed in claim 3 assembles the esterase catalyzed dose of application in biodiesel synthesis.
5. biology base metal organic frame as claimed in claim 4 assembles esterase catalyzed dose of answering in biodiesel synthesis
With, it is characterised in that: using sunflower oil as substrate, comprise the concrete steps that,
A) by right want 3 described in QLM@Bio-MOF 80~120mg of catalyst, ultrasonic disperse is in 20~40mL sunflower oil；
B) it is added methanol in Xiang Shangshu dispersion liquid, the volume ratio of sunflower oil and methanol is 2~8:1, and uses the NaOH tune of 1M
PH=8~10 for saving reaction solution, are then vigorously stirred 2~8h of lower reaction at 40~70 DEG C；
C) it is filtered to remove QLM@Bio-MOF catalyst, simultaneously rotary evaporation removing upper layer is clear using separatory funnel separation supernatant liquor
Unreacted methanol in liquid obtains biodiesel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811550027.2A CN109569728A (en) | 2018-12-18 | 2018-12-18 | A kind of biology base metal organic frame assembles esterase catalyzed dose, preparation method and its application in biodiesel synthesis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811550027.2A CN109569728A (en) | 2018-12-18 | 2018-12-18 | A kind of biology base metal organic frame assembles esterase catalyzed dose, preparation method and its application in biodiesel synthesis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109569728A true CN109569728A (en) | 2019-04-05 |
Family
ID=65930785
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811550027.2A Pending CN109569728A (en) | 2018-12-18 | 2018-12-18 | A kind of biology base metal organic frame assembles esterase catalyzed dose, preparation method and its application in biodiesel synthesis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109569728A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110368903A (en) * | 2019-06-10 | 2019-10-25 | 青岛职业技术学院 | The enrichment method of polyunsaturated fatty acid in a kind of grease |
| CN112725324A (en) * | 2020-12-31 | 2021-04-30 | 东莞市汇林包装有限公司 | Enzyme-based composite catalyst, preparation method and use method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104087572A (en) * | 2014-07-01 | 2014-10-08 | 清华大学 | Protein and metal organic skeleton compound composite material and preparation method thereof |
| CN105646253A (en) * | 2016-02-29 | 2016-06-08 | 华南理工大学 | Biocompatible metal organic framework material glutamic acid-zinc as well as preparation and application thereof |
| CN106715690A (en) * | 2014-07-03 | 2017-05-24 | 联邦科学和工业研究组织 | Host-guest metal organic framework systems |
| CN106755145A (en) * | 2016-12-21 | 2017-05-31 | 南京工业大学 | A kind of method of the fixed lipase catalyzed synthesizing citric acid ester of utilization nano composite material |
| CN108384025A (en) * | 2018-04-08 | 2018-08-10 | 东华大学 | A kind of biological MOF and preparation method thereof based on adenine |
| CN108396023A (en) * | 2018-02-09 | 2018-08-14 | 兰州大学 | Magnetism MOF materials are prepared with polishing and for the fixation of enzyme |
-
2018
- 2018-12-18 CN CN201811550027.2A patent/CN109569728A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104087572A (en) * | 2014-07-01 | 2014-10-08 | 清华大学 | Protein and metal organic skeleton compound composite material and preparation method thereof |
| CN106715690A (en) * | 2014-07-03 | 2017-05-24 | 联邦科学和工业研究组织 | Host-guest metal organic framework systems |
| CN105646253A (en) * | 2016-02-29 | 2016-06-08 | 华南理工大学 | Biocompatible metal organic framework material glutamic acid-zinc as well as preparation and application thereof |
| CN106755145A (en) * | 2016-12-21 | 2017-05-31 | 南京工业大学 | A kind of method of the fixed lipase catalyzed synthesizing citric acid ester of utilization nano composite material |
| CN108396023A (en) * | 2018-02-09 | 2018-08-14 | 兰州大学 | Magnetism MOF materials are prepared with polishing and for the fixation of enzyme |
| CN108384025A (en) * | 2018-04-08 | 2018-08-10 | 东华大学 | A kind of biological MOF and preparation method thereof based on adenine |
Non-Patent Citations (3)
| Title |
|---|
| FEREIDOON SHAHIDI等编著: "《贝雷油脂化学与工艺学》", 31 July 2016, 中国轻工业出版社 * |
| JYOTI SINGH ET AL.: ""Immobilized lipase from Schizophyllum commune ISTL04 for the production of fatty acids methyl esters from cyanobacterial oil"", 《BIORESOURCE TECHNOLOGY》 * |
| SHAMRAJA S. NADAR ET AL.: ""Encapsulation of lipase within metal-organic framework (MOF) with enhanced activity intensified under ultrasound"", 《ENZYME AND MICROBIAL TECHNOLOGY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110368903A (en) * | 2019-06-10 | 2019-10-25 | 青岛职业技术学院 | The enrichment method of polyunsaturated fatty acid in a kind of grease |
| CN112725324A (en) * | 2020-12-31 | 2021-04-30 | 东莞市汇林包装有限公司 | Enzyme-based composite catalyst, preparation method and use method thereof |
| CN112725324B (en) * | 2020-12-31 | 2023-10-20 | 广东省汇林包装科技集团有限公司 | Enzyme-based composite catalyst, preparation method and use method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Quayson et al. | Immobilized lipases for biodiesel production: Current and future greening opportunities | |
| Xie et al. | Enzymatic transesterification of soybean oil by using immobilized lipase on magnetic nano-particles | |
| Nezhad et al. | Tosylated cloisite as a new heterofunctional carrier for covalent immobilization of lipase and its utilization for production of biodiesel from waste frying oil | |
| Hu et al. | Rationally designing hydrophobic UiO-66 support for the enhanced enzymatic performance of immobilized lipase | |
| Bayramoglu et al. | Immobilized lipase on micro-porous biosilica for enzymatic transesterification of algal oil | |
| Dizge et al. | Biodiesel production from canola oil by using lipase immobilized onto hydrophobic microporous styrene–divinylbenzene copolymer | |
| Xie et al. | Immobilized lipase on magnetic chitosan microspheres for transesterification of soybean oil | |
| Li et al. | Preparation of immobilized lipase by modified polyacrylonitrile hollow membrane using nitrile-click chemistry | |
| Valério et al. | Understanding the biocatalytic potential of lipase from Rhizopus chinensis | |
| CN109706141B (en) | Immobilized enzyme pickering emulsion reaction system and application thereof | |
| CN105506013B (en) | A kind of method preparing duck oil diacylglycerol | |
| Jia et al. | Selective production of ethyl levulinate from levulinic acid by lipase-immobilized mesoporous silica nanoflowers composite | |
| Zhang et al. | Chemo-enzymatic epoxidation of Sapindus mukurossi fatty acids catalyzed with Candida sp. 99–125 lipase in a solvent-free system | |
| CN109569728A (en) | A kind of biology base metal organic frame assembles esterase catalyzed dose, preparation method and its application in biodiesel synthesis | |
| Abdulmalek et al. | Recent developments of lipase immobilization technology and application of immobilized lipase mixtures for biodiesel production | |
| Shalini et al. | Synthesis and characterisation of lipase immobilised magnetic nanoparticles and its role as a catalyst in biodiesel production | |
| CN111944798B (en) | Immobilized lipase, preparation method thereof and application thereof in biodiesel | |
| Wang et al. | Engineering balanced anions coupling with tailored functional groups of poly (ionic liquid) s immobilized lipase enables effective biodiesel production | |
| CN106591385B (en) | Method for preparing butyrin by enzyme method | |
| CN107586770A (en) | A kind of porcine pancreatic lipase inorganic hybrid nano flower catalyst, preparation method and its application in biodiesel preparation | |
| KR101548043B1 (en) | Preparation method of biodiesel using microorganism | |
| CN111378641A (en) | Immobilized enzyme carrier and immobilized enzyme | |
| Sundaramahalingam et al. | An encapsulated report on enzyme-assisted transesterification with an allusion to lipase | |
| Yuki et al. | Epoxidation of fatty acids by pluronic-conjugated lipase in organic media | |
| Zhang et al. | Lipase immobilization using scalable and biocompatible lignin-based material as a carrier |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190405 |