CN114014998A - A kind of method that utilizes soybean seed coat peroxidase to catalyze and synthesize copolymer - Google Patents
A kind of method that utilizes soybean seed coat peroxidase to catalyze and synthesize copolymer Download PDFInfo
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- CN114014998A CN114014998A CN202111462651.9A CN202111462651A CN114014998A CN 114014998 A CN114014998 A CN 114014998A CN 202111462651 A CN202111462651 A CN 202111462651A CN 114014998 A CN114014998 A CN 114014998A
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- coat peroxidase
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- 108010053276 anionic peroxidase Proteins 0.000 title claims abstract description 34
- 229920001577 copolymer Polymers 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 24
- 239000000178 monomer Substances 0.000 claims abstract description 82
- 238000006243 chemical reaction Methods 0.000 claims abstract description 69
- 239000012986 chain transfer agent Substances 0.000 claims abstract description 18
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000003999 initiator Substances 0.000 claims abstract description 14
- 239000007800 oxidant agent Substances 0.000 claims abstract description 13
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 claims abstract description 12
- 230000001590 oxidative effect Effects 0.000 claims abstract description 12
- 230000002194 synthesizing effect Effects 0.000 claims abstract description 8
- 239000012298 atmosphere Substances 0.000 claims abstract description 6
- 239000011261 inert gas Substances 0.000 claims abstract description 6
- 229920002554 vinyl polymer Polymers 0.000 claims abstract description 6
- 239000004721 Polyphenylene oxide Substances 0.000 claims abstract description 5
- 229920000570 polyether Polymers 0.000 claims abstract description 5
- 239000002904 solvent Substances 0.000 claims abstract description 5
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 claims abstract description 5
- 230000035484 reaction time Effects 0.000 claims abstract description 4
- 150000003926 acrylamides Chemical class 0.000 claims abstract description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 32
- YRKCREAYFQTBPV-UHFFFAOYSA-N acetylacetone Chemical compound CC(=O)CC(C)=O YRKCREAYFQTBPV-UHFFFAOYSA-N 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 16
- 229910052760 oxygen Inorganic materials 0.000 claims description 12
- 230000015572 biosynthetic process Effects 0.000 claims description 10
- 238000003786 synthesis reaction Methods 0.000 claims description 10
- OMIGHNLMNHATMP-UHFFFAOYSA-N 2-hydroxyethyl prop-2-enoate Chemical compound OCCOC(=O)C=C OMIGHNLMNHATMP-UHFFFAOYSA-N 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 8
- 238000010791 quenching Methods 0.000 claims description 8
- UUORTJUPDJJXST-UHFFFAOYSA-N n-(2-hydroxyethyl)prop-2-enamide Chemical compound OCCNC(=O)C=C UUORTJUPDJJXST-UHFFFAOYSA-N 0.000 claims description 7
- 125000004432 carbon atom Chemical group C* 0.000 claims description 4
- QNILTEGFHQSKFF-UHFFFAOYSA-N n-propan-2-ylprop-2-enamide Chemical compound CC(C)NC(=O)C=C QNILTEGFHQSKFF-UHFFFAOYSA-N 0.000 claims description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical group OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 3
- OMPJBNCRMGITSC-UHFFFAOYSA-N Benzoylperoxide Chemical compound C=1C=CC=CC=1C(=O)OOC(=O)C1=CC=CC=C1 OMPJBNCRMGITSC-UHFFFAOYSA-N 0.000 claims description 2
- 125000000217 alkyl group Chemical group 0.000 claims description 2
- 235000019400 benzoyl peroxide Nutrition 0.000 claims description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 2
- 229940088644 n,n-dimethylacrylamide Drugs 0.000 claims description 2
- YLGYACDQVQQZSW-UHFFFAOYSA-N n,n-dimethylprop-2-enamide Chemical compound CN(C)C(=O)C=C YLGYACDQVQQZSW-UHFFFAOYSA-N 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims description 2
- 125000005702 oxyalkylene group Chemical group 0.000 claims description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical group P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims 2
- 239000002953 phosphate buffered saline Substances 0.000 claims 2
- SBVKVAIECGDBTC-UHFFFAOYSA-N 4-hydroxy-2-methylidenebutanamide Chemical compound NC(=O)C(=C)CCO SBVKVAIECGDBTC-UHFFFAOYSA-N 0.000 claims 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 claims 1
- 102000003992 Peroxidases Human genes 0.000 claims 1
- 229920000359 diblock copolymer Polymers 0.000 claims 1
- 229920001519 homopolymer Polymers 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 229920006030 multiblock copolymer Polymers 0.000 claims 1
- 108040007629 peroxidase activity proteins Proteins 0.000 claims 1
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 claims 1
- 229910052939 potassium sulfate Inorganic materials 0.000 claims 1
- 235000011151 potassium sulphates Nutrition 0.000 claims 1
- 229920005604 random copolymer Polymers 0.000 claims 1
- 238000006116 polymerization reaction Methods 0.000 abstract description 20
- 102000004190 Enzymes Human genes 0.000 abstract description 12
- 108090000790 Enzymes Proteins 0.000 abstract description 12
- 239000003054 catalyst Substances 0.000 abstract description 12
- 238000006555 catalytic reaction Methods 0.000 abstract description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 abstract 2
- 238000005265 energy consumption Methods 0.000 abstract 1
- 230000002255 enzymatic effect Effects 0.000 abstract 1
- 239000008055 phosphate buffer solution Substances 0.000 description 20
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 239000005457 ice water Substances 0.000 description 12
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 10
- 239000001301 oxygen Substances 0.000 description 10
- 238000009826 distribution Methods 0.000 description 8
- 238000001914 filtration Methods 0.000 description 7
- 238000001542 size-exclusion chromatography Methods 0.000 description 7
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 6
- 238000001035 drying Methods 0.000 description 6
- 239000001257 hydrogen Substances 0.000 description 6
- 229910052739 hydrogen Inorganic materials 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 239000012299 nitrogen atmosphere Substances 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 230000001376 precipitating effect Effects 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- 238000001291 vacuum drying Methods 0.000 description 6
- 230000003197 catalytic effect Effects 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical group CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 4
- 238000007664 blowing Methods 0.000 description 3
- 238000007334 copolymerization reaction Methods 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 101150027801 CTA1 gene Proteins 0.000 description 2
- 101100273295 Candida albicans (strain SC5314 / ATCC MYA-2876) CAT1 gene Proteins 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 101100057143 Schizosaccharomyces pombe (strain 972 / ATCC 24843) cta3 gene Proteins 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- ZIWNJZLXPXFNGN-GXTQQWMXSA-N (z)-7-[(3r,4s)-3-[(e,3s)-3-hydroxyoct-1-enyl]-4-bicyclo[3.1.1]heptanyl]hept-5-enoic acid Chemical compound OC(=O)CCC\C=C/C[C@@H]1[C@@H](/C=C/[C@@H](O)CCCCC)CC2CC1C2 ZIWNJZLXPXFNGN-GXTQQWMXSA-N 0.000 description 1
- 101800000263 Acidic protein Proteins 0.000 description 1
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 1
- 102000008015 Hemeproteins Human genes 0.000 description 1
- 108010089792 Hemeproteins Proteins 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- ACIAHEMYLLBZOI-ZZXKWVIFSA-N Unsaturated alcohol Chemical compound CC\C(CO)=C/C ACIAHEMYLLBZOI-ZZXKWVIFSA-N 0.000 description 1
- QYKIQEUNHZKYBP-UHFFFAOYSA-N Vinyl ether Chemical compound C=COC=C QYKIQEUNHZKYBP-UHFFFAOYSA-N 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000011942 biocatalyst Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000007046 ethoxylation reaction Methods 0.000 description 1
- 229920001002 functional polymer Polymers 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 150000003278 haem Chemical class 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 238000005580 one pot reaction Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004032 porphyrins Chemical class 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 229950003776 protoporphyrin Drugs 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 238000010526 radical polymerization reaction Methods 0.000 description 1
- 238000012712 reversible addition−fragmentation chain-transfer polymerization Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F283/00—Macromolecular compounds obtained by polymerising monomers on to polymers provided for in subclass C08G
- C08F283/06—Macromolecular compounds obtained by polymerising monomers on to polymers provided for in subclass C08G on to polyethers, polyoxymethylenes or polyacetals
- C08F283/065—Macromolecular compounds obtained by polymerising monomers on to polymers provided for in subclass C08G on to polyethers, polyoxymethylenes or polyacetals on to unsaturated polyethers, polyoxymethylenes or polyacetals
-
- 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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes
-
- 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
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
-
- 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
- C12P7/62—Carboxylic acid esters
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- General Chemical & Material Sciences (AREA)
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- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明公开了一种利用大豆种皮过氧化物酶催化合成共聚物的方法，包括以下步骤：将单体1、单体2、链转移剂、大豆种皮过氧化物酶、引发剂、溶剂在惰性气体气氛条件下混合，预热，然后在惰性气体气氛条件下加入氧化剂，反应，即得；所述单体1选自丙烯酸或丙烯酰胺类化合物，所述单体2选自乙烯基聚醚类大分子单体。相较于传统聚合，本发明利用大豆种皮过氧化物酶催化剂进行酶催化过程，结合生物酶催化绿色、高效的优点，极大地提高了不活泼单体(乙烯基聚醚类大分子单体)的反应效率，产物中不同组分的链含量均接近理论值，减少了反应时间，使反应过程具有安全、高效、能耗低，反应速率快，反应条件温和等优点。The invention discloses a method for synthesizing a copolymer catalyzed by soybean seed coat peroxidase, comprising the following steps: synthesizing monomer 1, monomer 2, chain transfer agent, soybean seed coat peroxidase, initiator and solvent Mix under inert gas atmosphere, preheat, then add oxidant under inert gas atmosphere, react, and get; the monomer 1 is selected from acrylic acid or acrylamide compounds, and the monomer 2 is selected from vinyl polymer ether macromonomers. Compared with traditional polymerization, the present invention utilizes soybean seed coat peroxidase catalyst to carry out the enzymatic catalysis process, combined with the advantages of green and efficient catalysis of biological enzymes, and greatly improves the efficiency of inactive monomers (vinyl polyether macromonomers). ), the chain content of different components in the product is close to the theoretical value, which reduces the reaction time and makes the reaction process safe, efficient, low in energy consumption, fast in reaction rate, and mild in reaction conditions.
Description
Technical Field
The invention relates to polymer synthesis, in particular to a method for synthesizing a copolymer by utilizing soybean seed coat peroxidase catalysis.
Background
Copolymers are also known as interpolymers. Polymers obtained by the polymerization of two or more different monomers. It can combine the excellent properties of various polymers to obtain functional polymer material with excellent performance. The enzyme is a biocatalyst with high efficiency and specificity, and the enzyme catalysis has the characteristics of greenness, no toxicity, reproducibility, mild reaction conditions, environmental friendliness and the like. At present, the method for synthesizing the copolymer is various, and the enzyme catalyzes RAFT polymerization by a one-pot method without an intermediate purification step, so that one-step polymerization of the monomer is realized. However, there are two limitations to its preparation: 1) the reaction time is long, and the reaction condition is strict; 2) the inactive monomer is difficult to realize high-efficiency polymerization; 3) the cost of the biological enzyme is high, and the wide application of the enzyme is limited.
Vinyl ether macromonomer is the mainstream macromonomer product at present, and the macromonomer is mainly synthesized into polyethylene glycol ether with terminal group double bonds by ethoxylation reaction of micromolecule unsaturated alcohol initiators with different structures. However, the activity is low, and the polymerization efficiency in radical polymerization is not high.
Disclosure of Invention
The purpose of the invention is as follows: the invention provides a method for synthesizing a copolymer by using soybean seed coat peroxidase catalysis, which aims to overcome the defects of low reaction efficiency, low conversion rate and the like in the prior art.
The technical scheme is as follows: in order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a method for synthesizing a copolymer by using soybean seed coat peroxidase catalysis comprises the following steps:
mixing the monomer 1, the monomer 2, the chain transfer agent, the soybean seed coat peroxidase, the initiator and the solvent under the inert gas atmosphere condition, preheating, adding an oxidant under the inert gas atmosphere condition, and reacting to obtain the soybean seed coat peroxidase;
the monomer 1 is selected from acrylic acid or acrylamide compounds, and the monomer 2 is selected from vinyl polyether macromonomers.
Preferably, the monomer 1 is selected from N, N-Dimethylacrylamide (DMA), N-Hydroxyethylacrylamide (HEAA), Acrylic Acid (AA), hydroxyethyl acrylate (HEA) or N-isopropylacrylamide (NIPAM), and the structure is shown as follows.
Preferably, the monomer 2 is selected from vinyl polyether macromonomers, and the structural formula is as follows:
in the formula R1Is H or methyl, R2Is H or 1 &Alkyl of 4 carbon atoms, X ═ COO, O (CH)2)mO、CH2O or CH2CH2O and m are integers of 2-4; AO is selected from one or more of oxyalkylene groups with 2-4 carbon atoms, and n is the average addition mole number of AO and is an integer of 20-100; (AO)nCan be a homopolymerized structure, a random copolymerization structure, a diblock structure or a multiblock copolymerization structure. The method comprises the following specific steps:
preferably, the chain transfer agent CTA is selected from one or more of the following formulae:
preferably, the initiator is acetylacetone (ACAC), dibenzoyl peroxide (BOP) or potassium persulfate, and the structure is as follows:
preferably, the oxidizing agent is hydrogen peroxide (H)2O2)。
Preferably, the solvent is a mixed solution of Phosphate Buffer Solution (PBS)/dimethyl sulfoxide (DMSO), and the pH value is 6-8.
Preferably, the molar ratio of the monomer 1 to the chain transfer agent is 100-160: 1, the molar ratio of the chain transfer agent to the soybean seed coat peroxidase to the initiator to the oxidant is 1: 0.0005-0.001: 6:0.3, and the molar ratio of the monomer 1 to the monomer 2 is 2-5: 1.
Preferably, the reaction time is 30-120 min, and the reaction temperature is 25-35 ℃.
Preferably, after the reaction is finished, adding an organic solvent into the reaction solution, quenching, separating and purifying to obtain the copolymer of the macromonomer 1-the macromonomer 2.
The catalyst is soybean seed coat peroxidase, and the active center of the catalyst is shown as follows:
soybean seed coat peroxidase is a heme protein consisting of a single peptide chain and porphyrin iron, has a molecular weight of about 37KD and consists of more than 300 amino acid residues. The soybean seed coat peroxidase consists of two different structural domains, and a heme prosthetic group with a catalytic active center is embedded in the middle of the soybean seed coat peroxidase. It has an isoelectric point of 3.9 and belongs to acidic proteins. It contains Fe (III) -protoporphyrin IX prosthetic group, one tryptophan, 4 disulfide bonds, 2 Ca2+And 8 polysaccharides. Compared with the catalytic enzyme in the prior art, the catalytic enzyme has higher stability and catalytic activity.
The invention utilizes the advantages of the novel biological enzyme catalyst, leads the reaction to be mild and efficient, constructs a reaction unit aiming at the specific catalyst, the active monomer and the inactive monomer, and realizes the promotion of the polymerization reaction rate and the optimization of the molecular weight distribution; by means of the unique advantages of enzyme catalysis, the polymerization rate of the inactive monomer is improved, and the efficient copolymerization of different monomers is realized; provides a new technical reference for the polymerization of vinyl polyether macromonomer and other inactive monomers.
Has the advantages that: compared with the prior art, the method utilizes the process of the novel bio-enzyme catalysis, combines the advantages of the enzyme catalysis, reserves the catalytic efficiency of the catalyst on different monomers, improves the reaction rate, optimizes the process flow, and has the advantages of safety, high efficiency, greenness and controllable molecular weight; meanwhile, the soybean seed coat peroxidase is cheap and easy to obtain, has low cost and provides a basis for industrial production.
Detailed Description
The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.
In the following examples of the present invention, the molecular weight and molecular weight distribution of the product were measured by the following methods.
Using a Wyatt size exclusion chromatography system, a GPC column equipped with an SSI 1500 pump, a Wyatt OptilabEX detector, Waters Styragel HR;
analysis conditions were as follows: the mobile phase is N, N-dimethylformamide, the flow rate is 0.7mL/min, the column temperature is 25 ℃, and the sample injection volume is 0.4 mL.
Sample measurement: taking a pure sample of 2mg, adding 1mLN, N-dimethylformamide solution for dilution, filtering by using a disposable filter head, and taking 4mL of solution for sample measurement.
In the following examples of the present invention, the conversion C represents the molar ratio of the reacted monomers to the total amount of the starting monomers and can be calculated as follows:
C＝(na/n0)*100％
wherein C represents the conversion of the monomer, naDenotes the molar amount of monomer reacted, n0Representing the total molar amount of the initial monomers.
Example 1
Assuming that the molar ratio of the monomer 1 to the chain transfer agent to the catalyst to the initiator to the oxidant is 120:1:0.00081:6:0.3, the molar ratio of the monomer 1 to the monomer 2 is 3:1, the monomer 1 is DMA, and the monomer 2 is APEG2300. 12.26mg of CTA1, 6.56g of DMA, 39.25g of TPEG2300And a mixed solution of 250ml PBS and DMSO (PBS/DMSO 26:74v/v, [ PBS)]20mM, pH 6) is added into a reaction bottle and stirred evenly, 19.13mg of soybean seed coat peroxidase and 0.33g of ACAC are added into the reaction bottle, the reaction bottle is sealed and placed into an ice water bath for nitrogen blowing and oxygen discharging for 30min, then the reaction bottle is placed into an oil bath at the temperature of 30 ℃, after the temperature is stabilized, 100 mu L of deoxygenated H is put into a micro syringe2O2The solution was injected into a reaction flask to initiate polymerization. After 2h of polymerization under nitrogen atmosphere, the reaction flask was placed in an ice-water bath and oxygen was introduced to quench the reaction. Adding a large amount of methanol into the reaction solution, and stirringThen precipitating for 4h at low temperature, filtering, collecting precipitate, drying in a vacuum drying oven for 48h, and analyzing by size exclusion chromatography and nuclear magnetic hydrogen spectrum to obtain a result that the conversion rate of the high-activity monomer DMA is 98 percent and the conversion rate of the low-activity monomer APEG is low2300The conversion was 91%, and the number average molecular weight of the resulting copolymer was 89.26kg/mol, and the molecular weight distribution index was 1.02. Wherein the content of P (DMA) chain link is 83 percent, P (APEG)2300) The chain link content was 17%.
Example 2
Assuming that the molar ratio of the monomer 1 to the chain transfer agent to the catalyst to the initiator to the oxidant is 130:1:0.00081:6:0.3, the molar ratio of the monomer 1 to the monomer 2 is 4:1, the monomer 1 is HEAA, and the monomer 2 is APEG2300. 12.26mg of CTA1, 7.47g of HEAA, 37.37g of APEG2300And a mixed solution of 200ml PBS and DMSO (PBS/DMSO 26:74v/v, [ PBS)]20mM, pH 6) was added to the flask and stirred uniformly, 19.13mg of soybean seed coat peroxidase and 0.56g of BOP were added thereto, the flask was sealed, the flask was placed in an ice water bath and purged with nitrogen for 30min, then the flask was placed in an oil bath at 30 ℃ and, after the temperature stabilized, 100. mu.L of deoxygenated H was injected using a micro syringe2O2The solution was injected into a reaction flask to initiate polymerization. After 2h of polymerization under nitrogen atmosphere, the reaction flask was placed in an ice-water bath and oxygen was introduced to quench the reaction. Adding a large amount of methanol into the reaction solution, stirring, precipitating at low temperature for 4h, filtering, collecting precipitate, drying in a vacuum drying oven for 48h, and analyzing by size exclusion chromatography and nuclear magnetic hydrogen spectrum to obtain high-activity monomer HEAA with conversion rate of 98% and low-activity monomer APEG2300The conversion was 90%, the number-average molecular weight of the resulting copolymer was 92.18kg/mol, and the molecular weight distribution index was 1.09. Wherein the content of P (HEAA) chain unit is 88%, and P (APEG)2300) The chain link content was 12%.
Example 3
Assuming that the molar ratio of the monomer 1 to the chain transfer agent to the catalyst to the initiator to the oxidant is 130:1:0.0006:6:0.3, the molar ratio of the monomer 1 to the monomer 2 is 3:1, the monomer 1 is AA, and the monomer 2 is APEG4000. 12.26mg of CTA2, 4.6g of AA, 40.26g of APEG4000And a mixed solution of 250ml PBS and DMSO (PBS/DMSO 26:74v/v, [ PBS)]20mM, pH 7) was added to the reaction flask and stirred well, followed by addition of 19.1Sealing 3mg soybean seed coat peroxidase and 0.33g ACAC, placing the reaction bottle in ice water bath, blowing nitrogen to remove oxygen for 30min, placing in oil bath at 30 deg.C, after temperature is stable, using microinjector to remove oxygen 100 μ L H2O2The solution was injected into a reaction flask to initiate polymerization. After 2h of polymerization under nitrogen atmosphere, the reaction flask was placed in an ice-water bath and oxygen was introduced to quench the reaction. Adding a large amount of methanol into the reaction solution, stirring, precipitating at low temperature for 4h, filtering, collecting precipitate, drying in a vacuum drying oven for 48h, and analyzing by size exclusion chromatography and nuclear magnetic hydrogen spectrum to obtain a high-activity monomer AA conversion rate of 95% and a low-activity monomer APEG4000The conversion was 92%, the number average molecular weight of the resulting copolymer was 79.80kg/mol, and the molecular weight distribution index was 1.09. Wherein the content of P (AA) chain unit is 76 percent, and P (APEG)4000) The chain link content was 24%.
Example 4
Assuming that the molar ratio of the monomer 1 to the chain transfer agent to the catalyst to the initiator to the oxidant is 140:1:0.00081:6:0.3, the molar ratio of the monomer 1 to the monomer 2 is 5:1, the monomer 1 is HEA, and the monomer 2 is HPEG2300. 12.26mg of CTA, 7.58g of HEA, 34.86g of HPEG2300And a mixed solution of 200mL of PBS and DMSO (PBS/DMSO 26:74v/v, [ PBS)]20mM, pH 7) is added into a reaction bottle and stirred evenly, 19.13mg of soybean seed coat peroxidase and 0.33g of ACAC are added into the reaction bottle, the reaction bottle is sealed and placed into an ice water bath for nitrogen blowing and oxygen discharging for 30min, then the reaction bottle is placed into an oil bath at the temperature of 30 ℃, after the temperature is stabilized, 100 mu L of deoxygenated H is put into a micro syringe2O2The solution was injected into a reaction flask to initiate polymerization. After 2h of polymerization under nitrogen atmosphere, the reaction flask was placed in an ice-water bath and oxygen was introduced to quench the reaction. Adding a large amount of methanol into the reaction solution, stirring, precipitating at low temperature for 4h, filtering, collecting precipitate, drying in a vacuum drying oven for 48h, and analyzing by size exclusion chromatography and nuclear magnetic hydrogen spectrum to obtain monomer HPEG with high activity and high conversion rate of HEA 95% and low activity2300The conversion was 95%, the number average molecular weight of the resulting copolymer was 75.24kg/mol, and the molecular weight distribution index was 1.10. Wherein the content of P (HEA) chain unit is 86%, and P (HPEG)2300) The chain link content was 14%.
Example 5
Assuming that the molar ratio of the monomer 1 to the chain transfer agent to the catalyst to the initiator to the oxidant is 100:1:0.00081:6:0.3, the molar ratio of the monomer 1 to the monomer 2 is 4:1, the monomer 1 is DMA, and the monomer 2 is HPEG4000. 12.26mg of CTA3, 5.42g of DMA, 31.16g of HPEG4000And a mixed solution of 200ml PBS and DMSO (PBS/DMSO 26:74v/v, [ PBS)]20mM, pH 8) was added to the reaction flask and stirred uniformly, 19.13mg of soybean seed coat peroxidase and 0.33g of potassium persulfate were added thereto, the flask was sealed, the flask was placed in an ice water bath and purged with nitrogen for 30min, then the flask was placed in an oil bath at 30 ℃ and, after the temperature stabilized, 100. mu.L of deoxygenated H was put in an oil bath using a micro syringe2O2The solution was injected into a reaction flask to initiate polymerization. After 2h of polymerization under nitrogen atmosphere, the reaction flask was placed in an ice-water bath and oxygen was introduced to quench the reaction. Adding a large amount of methanol into the reaction solution, stirring, precipitating at low temperature for 4h, filtering, collecting precipitate, drying in a vacuum drying oven for 48h, and analyzing by size exclusion chromatography and nuclear magnetic hydrogen spectrum to obtain a low-activity monomer HPEG with high DMA conversion rate of 93% and high DMA conversion rate of high-activity monomer4000The conversion was 90%, the number average molecular weight of the resulting copolymer was 72.58kg/mol, and the molecular weight distribution index was 1.15. Wherein the content of P (DMA) chain links is 90 percent, and P (HPEG)4000) The chain link content was 10%.
Example 6
Assuming that the molar ratio of the monomer 1 to the chain transfer agent to the catalyst to the initiator to the oxidant is 100:1:0.001:6:0.3, the molar ratio of the monomer 1 to the monomer 2 is 4:1, the monomer 1 is AA, and the monomer 2 is TPEG2300. 12.26mg of CTA3, 4.5g of AA, 28.23g of TPEG2300And a mixed solution of 200mL of PBS and DMSO (PBS/DMSO 26:74v/v, [ PBS)]20mM, pH 8) was added to the reaction flask and stirred uniformly, 19.13mg of soybean seed coat peroxidase and 0.33g of ACAC were added thereto, the flask was sealed, the flask was placed in an ice water bath and purged with nitrogen for 30min, then the flask was placed in an oil bath at 30 ℃ and, after the temperature was stabilized, 100. mu.L of deoxygenated H was put in an oil bath using a micro syringe2O2The solution was injected into a reaction flask to initiate polymerization. After 2h of polymerization under nitrogen atmosphere, the reaction flask was placed in an ice-water bath and oxygen was introduced to quench the reaction. A large amount of methanol was added to the reaction solution,stirring, low-temperature precipitating for 4h, filtering, collecting precipitate, drying in vacuum drying oven for 48h, and analyzing by size exclusion chromatography and nuclear magnetic hydrogen spectrum to obtain high-activity monomer AA with conversion rate of 94% and low-activity monomer TPEG2300The conversion was 91%, the number-average molecular weight of the resulting copolymer was 58.54kg/mol, and the molecular weight distribution index was 1.12. Wherein the content of P (AA) chain unit is 83%, and P (TPEG)2300) The chain link content was 17%.
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| DE102009009209A1 (en) * | 2008-08-18 | 2010-02-25 | Basf Se | Preparing comb polymers comprises providing polymers having 1,3-dicarbonyl group and polymerization of ethylenically unsaturated monomer in presence of polymers in aqueous polymerization medium, initiated by peroxidase and peroxide source |
| CN103396515A (en) * | 2013-08-09 | 2013-11-20 | 西南石油大学 | High temperature resistant modified starch filtration reducing agent and preparation method thereof |
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| DE102009009209A1 (en) * | 2008-08-18 | 2010-02-25 | Basf Se | Preparing comb polymers comprises providing polymers having 1,3-dicarbonyl group and polymerization of ethylenically unsaturated monomer in presence of polymers in aqueous polymerization medium, initiated by peroxidase and peroxide source |
| CN103396515A (en) * | 2013-08-09 | 2013-11-20 | 西南石油大学 | High temperature resistant modified starch filtration reducing agent and preparation method thereof |
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