CN113801873B - Metal organic framework multienzyme composite catalytic material, preparation method, application and use method thereof - Google Patents
Metal organic framework multienzyme composite catalytic material, preparation method, application and use method thereof Download PDFInfo
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- CN113801873B CN113801873B CN202111103745.7A CN202111103745A CN113801873B CN 113801873 B CN113801873 B CN 113801873B CN 202111103745 A CN202111103745 A CN 202111103745A CN 113801873 B CN113801873 B CN 113801873B
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- 239000000463 material Substances 0.000 title claims abstract description 78
- 230000003197 catalytic effect Effects 0.000 title claims abstract description 53
- 239000012621 metal-organic framework Substances 0.000 title claims abstract description 40
- 239000002131 composite material Substances 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 108090000698 Formate Dehydrogenases Proteins 0.000 claims description 68
- 108010054269 Uridine Diphosphate Glucose Dehydrogenase Proteins 0.000 claims description 55
- 102000057144 Uridine Diphosphate Glucose Dehydrogenase Human genes 0.000 claims description 54
- 238000006243 chemical reaction Methods 0.000 claims description 46
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 45
- 239000000243 solution Substances 0.000 claims description 43
- HSCJRCZFDFQWRP-UHFFFAOYSA-N Uridindiphosphoglukose Natural products OC1C(O)C(O)C(CO)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-UHFFFAOYSA-N 0.000 claims description 37
- HDYANYHVCAPMJV-USQUEEHTSA-N udp-glucuronic acid Chemical compound O([P@](O)(=O)O[P@](O)(=O)OC[C@H]1[C@@H]([C@H]([C@@H](O1)N1C(NC(=O)C=C1)=O)O)O)[C@H]1O[C@@H](C(O)=O)[C@H](O)[C@@H](O)[C@@H]1O HDYANYHVCAPMJV-USQUEEHTSA-N 0.000 claims description 27
- HDYANYHVCAPMJV-UHFFFAOYSA-N Uridine diphospho-D-glucuronic acid Natural products O1C(N2C(NC(=O)C=C2)=O)C(O)C(O)C1COP(O)(=O)OP(O)(=O)OC1OC(C(O)=O)C(O)C(O)C1O HDYANYHVCAPMJV-UHFFFAOYSA-N 0.000 claims description 26
- ONDPHDOFVYQSGI-UHFFFAOYSA-N zinc nitrate Chemical compound [Zn+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ONDPHDOFVYQSGI-UHFFFAOYSA-N 0.000 claims description 22
- 239000007853 buffer solution Substances 0.000 claims description 21
- 102100039702 Alcohol dehydrogenase class-3 Human genes 0.000 claims description 20
- 108010051015 glutathione-independent formaldehyde dehydrogenase Proteins 0.000 claims description 20
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 16
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims description 16
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 claims description 16
- 239000000203 mixture Substances 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- HSCJRCZFDFQWRP-JZMIEXBBSA-N UDP-alpha-D-glucose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-JZMIEXBBSA-N 0.000 claims description 11
- 238000005530 etching Methods 0.000 claims description 11
- 238000004064 recycling Methods 0.000 claims description 11
- 239000000843 powder Substances 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- 238000005119 centrifugation Methods 0.000 claims description 9
- 239000013110 organic ligand Substances 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 8
- HPABFFGQPLJKBP-UHFFFAOYSA-N 5-fluoro-1H-pyrimidin-2-one Chemical compound OC1=NC=C(F)C=N1 HPABFFGQPLJKBP-UHFFFAOYSA-N 0.000 claims description 6
- PZKFSRWSQOQYNR-UHFFFAOYSA-N 5-methyl-1h-1,2,4-triazole Chemical compound CC1=NC=NN1 PZKFSRWSQOQYNR-UHFFFAOYSA-N 0.000 claims description 6
- 229920002683 Glycosaminoglycan Polymers 0.000 claims description 6
- 108010048916 alcohol dehydrogenase (acceptor) Proteins 0.000 claims description 6
- 239000002775 capsule Substances 0.000 claims description 6
- 239000003054 catalyst Substances 0.000 claims description 6
- 238000004817 gas chromatography Methods 0.000 claims description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 6
- 229920006395 saturated elastomer Polymers 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- LXBGSDVWAMZHDD-UHFFFAOYSA-N 2-methyl-1h-imidazole Chemical compound CC1=NC=CN1 LXBGSDVWAMZHDD-UHFFFAOYSA-N 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 239000000758 substrate Substances 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 230000002378 acidificating effect Effects 0.000 claims description 4
- 230000010355 oscillation Effects 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 3
- YAGCJGCCZIARMJ-UHFFFAOYSA-N N1C(=NC=C1)C=O.[Zn] Chemical compound N1C(=NC=C1)C=O.[Zn] YAGCJGCCZIARMJ-UHFFFAOYSA-N 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 3
- 229910052799 carbon Inorganic materials 0.000 claims description 3
- 239000013154 zeolitic imidazolate framework-8 Substances 0.000 claims description 3
- MFLKDEMTKSVIBK-UHFFFAOYSA-N zinc;2-methylimidazol-3-ide Chemical compound [Zn+2].CC1=NC=C[N-]1.CC1=NC=C[N-]1 MFLKDEMTKSVIBK-UHFFFAOYSA-N 0.000 claims description 3
- 101000944170 Homo sapiens Histone acetyltransferase KAT8 Proteins 0.000 claims description 2
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 claims description 2
- 238000005273 aeration Methods 0.000 claims description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N formaldehyde Substances O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 2
- 238000006386 neutralization reaction Methods 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- 239000013094 zinc-based metal-organic framework Substances 0.000 claims description 2
- 101000939529 Homo sapiens UDP-glucose 6-dehydrogenase Proteins 0.000 claims 9
- 102100029640 UDP-glucose 6-dehydrogenase Human genes 0.000 claims 9
- 230000000694 effects Effects 0.000 abstract description 12
- 125000004122 cyclic group Chemical group 0.000 abstract description 4
- 230000002194 synthesizing effect Effects 0.000 abstract description 4
- 230000008929 regeneration Effects 0.000 abstract description 3
- 238000011069 regeneration method Methods 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 66
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 66
- 102000004190 Enzymes Human genes 0.000 description 40
- 108090000790 Enzymes Proteins 0.000 description 40
- 230000015572 biosynthetic process Effects 0.000 description 22
- 238000003786 synthesis reaction Methods 0.000 description 22
- 229910021642 ultra pure water Inorganic materials 0.000 description 14
- 239000012498 ultrapure water Substances 0.000 description 14
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 12
- 239000011701 zinc Substances 0.000 description 9
- 238000006555 catalytic reaction Methods 0.000 description 8
- 238000010586 diagram Methods 0.000 description 8
- 239000002244 precipitate Substances 0.000 description 8
- 238000010531 catalytic reduction reaction Methods 0.000 description 7
- 229910052751 metal Inorganic materials 0.000 description 7
- 239000002184 metal Substances 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 239000001569 carbon dioxide Substances 0.000 description 6
- 229910002092 carbon dioxide Inorganic materials 0.000 description 6
- 239000005515 coenzyme Substances 0.000 description 5
- LVRFTAZAXQPQHI-UHFFFAOYSA-N 2-hydroxy-4-methylvaleric acid Chemical compound CC(C)CC(O)C(O)=O LVRFTAZAXQPQHI-UHFFFAOYSA-N 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 239000000348 glycosyl donor Substances 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 229910052725 zinc Inorganic materials 0.000 description 3
- NOQGZXFMHARMLW-UHFFFAOYSA-N Daminozide Chemical compound CN(C)NC(=O)CCC(O)=O NOQGZXFMHARMLW-UHFFFAOYSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- XCCTYIAWTASOJW-XVFCMESISA-N Uridine-5'-Diphosphate Chemical compound O[C@@H]1[C@H](O)[C@@H](COP(O)(=O)OP(O)(O)=O)O[C@H]1N1C(=O)NC(=O)C=C1 XCCTYIAWTASOJW-XVFCMESISA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 230000000415 inactivating effect Effects 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000006722 reduction reaction Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- XYHKNCXZYYTLRG-UHFFFAOYSA-N 1h-imidazole-2-carbaldehyde Chemical compound O=CC1=NC=CN1 XYHKNCXZYYTLRG-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 108700023372 Glycosyltransferases Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000589774 Pseudomonas sp. Species 0.000 description 1
- 101150050063 UGDH gene Proteins 0.000 description 1
- 229910021536 Zeolite Inorganic materials 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 230000002785 anti-thrombosis Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000004146 energy storage Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000002803 fossil fuel Substances 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 102000045442 glycosyltransferase activity proteins Human genes 0.000 description 1
- 108700014210 glycosyltransferase activity proteins Proteins 0.000 description 1
- 239000005431 greenhouse gas Substances 0.000 description 1
- 150000002402 hexoses Chemical class 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000012924 metal-organic framework composite Substances 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 150000002972 pentoses Chemical class 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 230000009919 sequestration Effects 0.000 description 1
- 239000010457 zeolite Substances 0.000 description 1
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/008—Supramolecular polymers
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- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/098—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer formed in the presence of the enzymes or microbial cells
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
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- C12P7/00—Preparation of oxygen-containing organic compounds
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- C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
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- C12Y102/01046—Formaldehyde dehydrogenase (1.2.1.46)
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Abstract
The invention discloses a metal organic framework multienzyme composite catalytic material, a preparation method, application and a using method thereof. The composite catalytic material of the invention can effectively recycle CO 2 The method for solving the greenhouse effect is provided, and the method can realize repeated utilization for multiple times through immobilization and cyclic regeneration of cofactors while maintaining higher catalytic activity, and the method for synthesizing the high-value industrialized product with low cost and environmental protection is provided.
Description
Technical Field
The invention relates to a catalytic material, a preparation method, application and a using method, in particular to a metal organic framework multienzyme composite catalytic material, a preparation method, application and a using method thereof.
Background
Glycosaminoglycans are a class of linear polysaccharides that are widely found on animal cell surfaces and intercellular matrices, and are complex macromolecules of carbohydrates. Glycosaminoglycans are also associated with a number of important diseases such as inflammation, tumors, cancers, and the like. In addition to being an important functional substance in organisms, it is also an important class of biopharmaceuticals, such as antithrombotic, treatment of arthritis, etc. The most common glycosyl donor in organisms is UDP-sugar, where UDP-glucose is the basic raw material forming other UDP-monosaccharides, and UDP-glucuronic acid is the key substrate for the conversion of UDP-sugar from hexose to pentose. Uridine Diphosphate (UDP) -glucuronic acid is a fundamental building block in glycosaminoglycan synthesis, and is significant in glycosaminoglycan synthesis.
With the massive consumption of fossil fuels and the gradual decrease of green vegetation, the concentration of carbon dioxide in the atmosphere increases dramatically, and carbon dioxide becomes a major greenhouse gas. Currently, carbon dioxide capture and sequestration techniques are efficient methods for large-scale centralized processing of carbon dioxide, but require subsequent desorption. The problems of energy consumption and high cost in the processes of compression, storage and the like are main obstacles of industrial practice. On the other hand, carbon dioxide is used as C which is rich in energy storage, safe and renewable on earth 1 The resource can be prepared into useful materials and chemical products through chemical conversion reaction, waste is changed into valuable, high-value utilization of the waste is realized, and chemical energy products such as formic acid, formaldehyde, methanol and the like are obtained.
At present, the most common methods for reducing carbon dioxide and synthesizing UDP-glucuronic acid at home and abroad are chemical synthesis and enzymatic synthesis. The chemical method has the characteristics that the synthesis steps are too complex, the reagents are not environment-friendly, the enzymatic synthesis is environment-friendly and environment-friendly compared with the chemical method, the reaction conditions of the enzymatic synthesis are milder, the harm to the environment is small, and the like, but the free enzyme is used for catalysis, the existing enzyme is dissolved in water and the activity is easily influenced by the environment, the collection and repeated utilization of the enzyme are difficult, the activity of the enzyme is difficult to maintain for repeated utilization, the catalysis cost is high, and the enzyme needs to be protected to maintain the biological activity.
Disclosure of Invention
The invention aims to: the invention aims to provide a metal-organic framework multienzyme composite catalytic material, which solves the problem that enzymes are difficult to collect for repeated use, and simultaneously reduces the cost of continuous enzyme catalysis, reduces the influence of environment on enzyme activity and is difficult to collect for repeated use. The invention also aims at providing a preparation method of the composite catalytic material. It is also an object of the present invention to provide a method of using and using the composite catalytic material.
The technical scheme is as follows: the metal organic framework multienzyme composite catalytic material comprises formate dehydrogenase, formaldehyde dehydrogenase, methanol dehydrogenase, UDP-glucose dehydrogenase and MOF material, wherein the composite catalytic material is provided with a hollow cavitation capsule, the formate dehydrogenase, the formaldehyde dehydrogenase and the methanol dehydrogenase are wrapped in the capsule, and the UDP-glucose dehydrogenase is fixed in the MOF material.
The metal organic framework multienzyme composite catalytic material comprises a MOF material which is a Zn-based MOFs material: ZIF-8, ZIF-90 or MAF-7; preferably, the molar ratio of zinc to organic ligand in ZIF-8 synthesis is 1:12-1:16, the molar ratio of zinc to organic ligand in ZIF-90 synthesis is 1:2-1:6, and the molar ratio of zinc to organic ligand in MAF-7 synthesis is 1:2-1:4.
The metal organic frame FDH/F ald The preparation method of the DH/ADH/UGDH@MOF composite catalytic material comprises the following steps: firstly, immobilizing formate dehydrogenase, formaldehyde dehydrogenase and methanol dehydrogenase by etching template ZPF to obtain FDH/F ald DH/ADH@ZPF solids followed by FDH/F pairs using MOFs material ald Co-immobilization of DH/ADH@ZPF and UDP-glucose dehydrogenase to give (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF, and then taking the ZPF as a sacrificial template by utilizing the difference of the ZPF and the MOF to obtain the medium cavitation metal-organic framework FDH/F ald DH/ADH/UGDH@MOF composite catalytic material.
The metal organic frame FDH/F ald The etched template ZPF is a zeolite pyrimidine framework material.
The metal organic frame FDH/F ald The preparation method of the DH/ADH/UGDH@MOF composite catalytic material specifically comprises the following steps:
(1) Preparation of FDH/F ald DH/ADH@ZPF: FDH, F ald DH. ADH is put into Tris-HCl buffer solution, then 2-hydroxy-5 fluoropyrimidine is added, finally zinc nitrate is added, and the mixture is stirred for a period of time, centrifuged, washed and dried to obtain solid;
(2) Preparation (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF: the obtained FDH/F ald DH/ADH@ZPF and UGDH are added into Tris-HCl buffer solution together, then organic ligand is added, zinc nitrate is added after stirring, after stirring reaction for a period of time, the solid is obtained through centrifugation, washing and drying;
(3) Preparation of FDH/F by etching ZPF ald DH/ADH/UGDH@MOF: the prepared (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF powder is added into an acidic buffer solution to be stirred, and after a period of time, the mixture is centrifuged, washed and dried to obtain a solid.
The metal organic frame FDH/F ald Preparation method of DH/ADH/UGDH@MOF composite catalytic material, wherein FDH and F are prepared in the step (1) ald DH. ADH concentration of 0.3-0.5 mg/mL, tris-HCl buffer pH of 7-8, concentration of 0.1-0.3M, 2-hydroxy-5 fluoropyrimidine concentration of 0.2-0.4M, zinc nitrate concentration of 0.05-0.1M, reaction at 35-45 ℃ and 200-400 rpm for 8-12 h, and drying temperature of 35-40 ℃; the concentration of UGDH in the step (2) is 0.2-0.5mg/mL, the pH of Tris-HCl buffer solution is 7-8, the concentration is 0.1-0.3M, the organic ligand is one of 2-formaldehyde imidazole, 2-methyl imidazole and 3-methyl-1,2,4-triazole, zinc nitrate is 30-50 mM, the reaction is carried out at 35-45 ℃ for 12-24 h at 200-400 rpm, and the drying temperature is 35-40 ℃; the acidic buffer solution in the step (3) is PB buffer solution, the pH is 3.5-5.5, the concentration is 50-70 mM, the etching time is 0.5-1 h, and the drying temperature is 35-40 ℃.
The metal organic frame FDH/F ald Use of DH/ADH/ugdh@mof composite catalytic material in carbon neutralization.
The metal organic frame FDH/F ald Application of DH/ADH/UGDH@MOF composite catalytic material in glycosaminoglycan production.
The metal organic frame FDH/F ald The application method of the DH/ADH/UGDH@MOF composite catalytic material comprises the following steps:
(1) CO is processed by 2 Continuously introducing pure water and preparing NADH solution;
(2) To be saturated with CO 2 After the solution preparation is completed, the freshly prepared NADH solution is added with CO 2 Adding UDP-glucose and FDH/F into the solution ald DH/ADH/UGDH@MOF composite catalytic material and sealingStirring uniformly, and carrying out oscillation reaction;
(3) Centrifuging the reaction solution in the step (2), collecting the composite catalyst after centrifugation, washing for recycling, taking the supernatant of the centrifugate, and detecting the generated methanol and UDP-glucuronic acid by a gas chromatography method.
The method of use, step (1) the CO 2 The aeration time is 30-50 min, and the NADH concentration is 30-50 mM; step (2) the F ald The adding amount of the DH/ADH/UGDH@MOF composite catalytic material is 5% -10% of the mass of the substrate; the oscillating reaction is a water bath oscillating reaction at 20-50 ℃, the rotating speed is 100-300 rpm, and the reaction time is 10-24 hours; the centrifugation condition in the step (3) is 8,000-12,000 rpm for 4-6 min.
Wherein, the catalytic reaction process of the enzyme composite catalytic material in the step (2) is shown in figure 1. In the case of CO 2 Catalyzing and reducing to methanol and catalyzing synthesis of UDP-glucuronic acid and catalyzing CO 2 The reduction of the cofactor NADH will oxidize NAD + And the synthesis of UDP-glucuronic acid leads to NAD + The NADH is reduced to form the cyclic regeneration of the coenzyme factors, and the production cost is further reduced while the efficient production is realized.
Specifically, the preparation of the present invention preferably comprises three parts:
a first part: FDH/F ald Preparation of DH/ADH@ZPF
FDH, F ald DH、ADH(FDH、F ald DH. ADH concentration is 0.3-0.5 mg/mL, tris-HCl buffer solution (pH=7-8,0.1-0.3M) is added, 2-hydroxy-5 fluoropyrimidine (0.2-0.4M) is added, zinc nitrate (0.05-0.1M) is added, and the mixture is stirred uniformly and then reacted at 35-45 ℃ for 8-12 h. After the reaction, the mixture is centrifuged at 8,000 to 12,000rpm for 4 to 6 minutes, and the precipitate is recovered. Then washed with ultrapure water, sonicated and centrifuged three times to remove free enzyme not encapsulated by the ZPF material. The method comprises the steps of carrying out a first treatment on the surface of the
A second part: (FDH/F) ald Preparation of DH/ADH@ZPF)/UGDH@MOF
1、(FDH/F ald DH/ADH@ZPF)/UGDH@ZIF-8
Taking 0.3mg FDH/F prepared ald DH/ADH@ZAdding 1mL Tris-HCl buffer solution into PFs powder, stirring, adding into 2-methyl-imidazole (HmIm, 640-750 mM) solution, adding Zn (NO) 3 ) 2 ·6H 2 O (40-50 mM) solution and adding ultrapure water to a constant volume of 3-5 mL. Reacting for 12-24 h at 37-45 ℃ and 300-500 rpm. After the reaction, the mixture is centrifuged at 8,000 to 12,000rpm for 4 to 6 minutes, and the precipitate is recovered. Then washed with ultrapure water, sonicated and centrifuged three times.
2、(FDH/F ald DH/ADH@ZPF)/UGDH@ZIF-90
Taking 0.3mg FDH/F prepared ald Adding DH/ADH@ZPFs powder into 1mL Tris-HCl buffer solution, stirring, adding into 2-imidozole-carboxaldehyde (HICA, 160-260 mM) solution, and adding Zn (NO) 3 ) 2 ·6H 2 O (40-50 mM) solution and adding ultrapure water to a constant volume of 3-5 mL. Reacting for 12-24 h at 37-45 ℃ and 200-300 rpm. After the reaction, the mixture is centrifuged at 8,000 to 12,000rpm for 4 to 6 minutes, and the precipitate is recovered. Then washed with ultrapure water, sonicated and centrifuged three times.
3、(FDH/F ald DH/ADH@ZPF)/UGDH@MAF-7
Taking 0.3mg FDH/F prepared ald DH/ADH@ZPFs powder is added into 1mL Tris-HCl buffer solution, stirred and mixed uniformly, and then added into 3-methyl-1,2,4-triazole (Hmtz, 120-200 mM) solution, and 10% NH is preferably added when MNPs-lip@MAF-7 is prepared 3 ·H 2 O (60-80 mu L) and Zn (NO) 3 ) 2 ·6H 2 O (40-60 mM) solution and ultrapure water are added to a constant volume of 3-5 mL. Reacting for 12-24 h at 37-5 ℃ and 300-500 rpm. After the reaction, the mixture is centrifuged at 8,000 to 12,000rpm for 4 to 6 minutes, and the precipitate is recovered. Then washed with ultrapure water, sonicated and centrifuged three times.
Third section: etching and FDH/F of template ZPF ald Preparation of DH/ADH/UGDH@MOF
The prepared (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF powder is added into PB (pH=3.5-5.5, 50-70 mM) buffer solution to be stirred, and after 0.5-1 h, the mixture is centrifuged (8,000-10,000 rpm, 5-8 min), washed and dried at 35-40 ℃ to obtain a solid.
Fourth partThe method comprises the following steps: FDH/F ald DH/ADH/UGDH@MOF composite catalytic material for catalytic reduction of CO 2 Synthesizing UDP-glucuronic acid.
CO is processed by 2 Introducing the mixture into pure water for 30-50 min, and simultaneously preparing an NADH (30-50 mM) solution. To be saturated with CO 2 After the solution preparation is completed, the freshly prepared NADH solution is added with CO 2 Adding FDH/F with the mass of 5-10% of that of 1-3 mM FDP-glucose and UDP-glucose into the solution in sequence ald DH/ADH/UGDH@MOF composite catalytic material is uniformly stirred after sealing, water bath is carried out at 20-50 ℃, and oscillating reaction is carried out at 100-300 rpm. And centrifuging the reaction solution for 10-24 hours under the conditions of 8,000-12,000 rpm for 4-6 minutes, collecting the composite catalyst after centrifugation, washing for recycling, taking the supernatant of the centrifugate, and detecting the generated methanol and UDP-glucuronic acid by a gas chromatography.
The FDH/F is realized by adopting the combined process ald DH/ADH is encapsulated in the ZPF post-etched pocket and UGDH is immobilized in the MOF. The influence of the catalytic environment on the enzyme is reduced, the mechanical property of the enzyme is enhanced, and the operation stability is improved. And the pore diameter of the MOF becomes larger by acid etching, so that the contact of the substrate and the enzyme is easier, and the mass transfer resistance is reduced.
The composite catalytic material of the invention can efficiently catalyze CO 2 The reduction reaction of (2) and the synthesis of UDP-glucuronic acid not only have good thermal stability and chemical stability, but also can improve the enzymatic reaction rate, and can realize the cyclic regeneration of coenzyme so as to further reduce the cost.
The innovation point of the invention is that (1) FDH/F is immobilized by ZPF ald DH/ADH, and form the larger capsule cavity through etching ZPF, reduce the possibility of inactivating denaturation of multienzyme in the material. (2) 3 MOFs which are suitable for effectively wrapping enzymes and are stable are found from a plurality of MOF materials, and the MOFs have mild synthesis conditions, do not need high temperature and high pressure, have low raw material cost and are simple in synthesis process. (3) UGDH is immobilized through a relatively stable MOF material, so that cascade catalysis of tetraase is realized. (4) The synthesis of the important glycosyl donor UDP-glucuronic acid is realized. (5) Realize the greenhouse effect gas CO 2 Is recycled to solve the greenhouse effectA solution is provided. (6) Realizes the cyclic utilization of the coenzyme factors and greatly reduces the production cost.
In addition, the MOF material is used for wrapping the tetraenzyme, protecting the enzyme from the influence of external environment, ensuring that the enzyme can resist the change of temperature, PH and organic solvent, maintaining the activity of the enzyme, improving the recycling rate of the enzyme, and being particularly capable of being recycled in UDP-glucuronic acid synthesis and CO 2 Reducing the cost of carbon recycle.
The invention uses Metal-organic frame materials (Metal-Organic Frameworks, MOFs), which are two-dimensional or three-dimensional crystal structures formed by self-assembly between Metal ions and organic ligands by taking the Metal ions as connection points and organic ligands as supports. In the enzyme catalytic reaction, enzymes are wrapped and fixed by metal organic framework materials, and the formation of the enzyme-metal organic framework material compound has the following advantages compared with free enzymes: in the reaction process, the method can resist a certain degree of denaturation conditions such as temperature, pH, organic solvents and the like; the porous nature of the metal organic framework material may promote contact of the enzyme with the substrate, increasing the reaction rate. The immobilized multienzyme is more beneficial to simulating the cell environment and realizing the synthesis of complex products on the basis of the metal organic framework material compound.
The beneficial effects are that: compared with the prior art, the invention has the following remarkable advantages: (1) The method forms a larger capsule cavity by etching the ZPF, thereby reducing the possibility of inactivating and denaturing the multienzyme in the material. The composite catalytic material is a stable material with high porosity, high specific surface area and adjustable structure, after the composite catalytic material is formed by wrapping multiple enzymes, the original high-efficiency, mild and specific enzyme catalytic activity of the enzymes can be maintained to a great extent, meanwhile, the defect of free enzymes is overcome, the storage stability of the enzymes is improved, the subsequent recovery is easy, the recovery step of the composite catalytic material is simplified, the recycling rate is improved, and the reaction cost can be reduced by realizing the coenzyme factor circulation. (2) The invention utilizes the designed novel immobilization material to immobilize UGDH, thereby realizing the synthesis of important glycosyl donor UDP-glucuronic acid. (3) The invention is used for immobilization of UGDHImmobilization of FDH/F ald DH/ADH, can enable CO 2 Catalytic reduction of generated NAD + The coenzyme factor is recycled, and the CO can be realized while the cost is reduced 2 Is a catalytic reduction of (a).
Drawings
FIG. 1 is a schematic diagram of a catalytic reaction process of an enzyme composite catalytic material;
FIG. 2 is an FDH/F ald DH/ADH/UGDH@ZIF-8 catalyzed CO 2 A conversion rate and a recycling rate of the generated methanol are shown in a schematic diagram;
FIG. 3 is an FDH/F ald DH/ADH/UGDH@ZIF-90 catalyzed CO 2 A conversion rate and a recycling rate of the generated methanol are shown in a schematic diagram;
FIG. 4 is an FDH/F ald DH/ADH/UGDH@MAF-7 catalyzed CO 2 A conversion rate and a recycling rate of the generated methanol are shown in a schematic diagram;
FIG. 5 is an FDH/F ald The conversion rate and the recycling rate of the UDP-glucuronic acid generated by catalyzing UDP-glucose by DH/ADH/UGDH@ZIF-8 are shown in the schematic diagram;
FIG. 6 is an FDH/F ald The conversion rate and the recycling rate of the UDP-glucuronic acid generated by catalyzing UDP-glucose by DH/ADH/UGDH@ZIF-90 are shown in the schematic diagram;
FIG. 7 is an FDH/F ald Conversion rate and recycling rate of UDP-glucuronic acid generated by catalyzing UDP-glucose by DH/ADH/UGDH@MAF-7 are shown in the schematic diagram;
FIG. 8 is a schematic diagram of enzyme activity in different environments of the composite catalytic material and free enzyme.
Detailed Description
Formate Dehydrogenase (FDH), formaldehyde dehydrogenase (F) ald DH), alcohol Dehydrogenase (ADH), UDP-glucose dehydrogenase (UGDH) is purified by self-expression of formate dehydrogenase (FDH, fromCandida boidinii), formaldehyde dehydrogenase (F) ald DH, from Pseudomonas sp.), alcohol dehydrogenase (ADH, fromSaccharomyces cerevisiae), UDP-glucose dehydrogenase (UGDH, from E.coli BL21 (DE 3)); UDP-glucose (purchased from Arraga Ding Shiji Co., ltd.); zinc nitrate (purchased from alar Ding Shiji limited), 2-methyl-imidozole (2-methylimidazole, purchased from alar Ding Shiji limited), imidozole-2-carboxaldihyde (imidazole-2-carbaldehyde, purchased from alaa Ding Shiji limited), 3-methyl-1,2,4-triazole (3-methyl-1, 2,4-triazole, purchased from alaa Ding Shiji limited).
Example 1
FDH/F ald Preparation of DH/ADH@ZPF:
FDH, F ald DH、ADH(FDH、F ald DH. ADH concentrations were 0.3mg/mL, tris-HCl buffer (pH= 7,0.1M) was added, 2-hydroxy-5 fluoropyrimidine (0.2M) was added, and finally zinc nitrate (0.05M) was added and stirred well before reacting at 35℃for 12h. After the reaction, the mixture was centrifuged at 8000rpm for 4 minutes to collect a precipitate. Then washed with ultrapure water, sonicated and centrifuged three times to remove free enzyme not encapsulated by the ZPF material.
Example 2
(FDH/F ald Preparation of DH/ADH@ZPF)/UGDH@MOF:
1、(FDH/F ald DH/ADH@ZPF)/UGDH@ZIF-8
taking 0.3mg FDH/F prepared ald DH/ADH@ZPFs powder is added into 1mL Tris-HCl buffer solution, stirred and mixed uniformly, added into 2-methyl-imidazole (HmIm, 640 mM) solution, and then added with Zn (NO) 3 ) 2 ·6H 2 O (40 mM) solution and ultrapure water were added to a constant volume of 3mL. The reaction was carried out at 37℃and 300rpm for 24 hours. After the completion of the reaction, the mixture was centrifuged at 8,000rpm for 4 minutes, and the precipitate was recovered. Then washed with ultrapure water, sonicated and centrifuged three times.
2、(FDH/F ald DH/ADH@ZPF)/UGDH@ZIF-90
Taking 0.3mg FDH/F prepared ald DH/ADH@ZPFs powder is added into 1mL Tris-HCl buffer solution, stirred and mixed uniformly, added into 2-imidozole-carboxaldyde (HICA, 160 mM) solution, and then added with Zn (NO) 3 ) 2 ·6H 2 O (40 mM) solution and ultrapure water were added to a constant volume of 3mL. The reaction was carried out at 37℃and 200rpm for 24 hours. After the reaction, the mixture was centrifuged at 8000rpm for 4 minutes to collect a precipitate. Then washed with ultrapure water, sonicated and centrifuged three times.
3、(FDH/F ald DH/ADH@ZPF)/UGDH@MAF-7
Taking 0.3mg FDH/F prepared ald DH/ADH@ZPFs powder is added into 1mL Tris-HCl buffer solution, stirred and mixed uniformly, then added into 3-methyl-1,2,4-triazole (Hmtz, 120 mM) solution, and 10% NH is preferably added when MNPs-lip@MAF-7 is prepared 3 ·H 2 O (60. Mu.L) and Zn (NO) 3 ) 2 ·6H 2 O (40 mM) solution and ultrapure water were added to a constant volume of 3mL. The reaction was carried out at 37℃and 500rpm for 24 hours. After the reaction, the mixture was centrifuged at 8000rpm for 4 minutes to collect a precipitate. Then washed with ultrapure water, sonicated and centrifuged three times.
Example 3
Etching and FDH/F of template ZPF ald Preparation of DH/ADH/UGDH@MOF:
the prepared (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF powder was added to PB (pH=5, 50 mM) buffer solution and stirred, after 0.5h, centrifuged (8000 rpm,5 min), washed and dried at 35℃to obtain a solid.
Example 4
(1)FDH/F ald DH/ADH/UGDH@ZIF-8 composite catalytic material for catalytic reduction of CO 2 Synthesis of UDP-glucuronic acid:
CO is processed by 2 A solution of NADH (30 mM) was prepared by passing it into pure water for 30 min. To be saturated with CO 2 After the solution preparation is completed, the freshly prepared NADH solution is added with CO 2 Adding FDH/F with mass of 1mM FDP-glucose and 5% UDP-glucose into the solution ald DH/ADH/UGDH@ZIF-8 composite catalytic material is uniformly stirred after sealing, water bath is carried out at 37 ℃, and shaking reaction is carried out at 200 rpm. After 12 hours, the reaction solution was centrifuged at 8000rpm for 4 minutes, and after centrifugation, the composite catalyst was collected and washed for reuse, and the supernatant of the centrifugate was collected and detected by gas chromatography to give methanol and UDP-glucuronic acid as shown in FIG. 2 and FIG. 5.
(2)FDH/F ald DH/ADH/UGDH@ZIF-90 composite catalytic material for catalytic reduction of CO 2 Synthesis of UDP-glucuronic acid:
CO is processed by 2 A solution of NADH (30 mM) was prepared by passing it into pure water for 30 min. To be saturated with CO 2 After the solution preparation is completed, the freshly prepared NADH solution is added with CO 2 Adding 1mM FDP-glucose and UDP-glucose into the solutionFDH/F of 10% by mass ald DH/ADH/UGDH@MOF composite catalytic material is uniformly stirred after sealing, water bath is carried out at 37 ℃, and oscillation reaction is carried out at 100 rpm. After 10 hours, the reaction solution was centrifuged at 8000rpm for 4 minutes, and after centrifugation, the composite catalyst was collected and washed for reuse, and the supernatant of the centrifugate was collected and detected by gas chromatography to obtain methanol and UDP-glucuronic acid as shown in FIG. 3 and FIG. 6.
(3)FDH/F ald DH/ADH/UGDH@MAF-7 composite catalytic material for catalytic reduction of CO 2 Synthesis of UDP-glucuronic acid:
CO is processed by 2 A solution of NADH (30 mM) was prepared by passing it into pure water for 30 min. To be saturated with CO 2 After the solution preparation is completed, the freshly prepared NADH solution is added with CO 2 Adding FDH/F with mass of 8% of 1mM FDP-glucose and UDP-glucose into the solution ald DH/ADH/UGDH@MOF composite catalytic material is uniformly stirred after sealing, water bath is carried out at 37 ℃, and oscillation reaction is carried out at 300 rpm. After 14 hours, the reaction solution was centrifuged at 8000rpm for 4 minutes, and after centrifugation, the composite catalyst was collected and washed for reuse, and the supernatant of the centrifugate was collected and detected by gas chromatography to give methanol and UDP-glucuronic acid as shown in FIG. 4 and FIG. 7.
Example 5
The enzyme activities of the different metal organic framework composite catalytic materials in example 4 of the present invention were tested under different environments and compared with the enzyme activities of free glycosyltransferases not encapsulated by MOFs material:
the three composite catalytic materials of example 4 and the free enzyme were treated in solutions of different temperatures, organic solvents and different pH for 0.5 hours, respectively. Three treated composite catalytic materials and free enzyme are used for catalytic reduction of CO 2 And synthesizing UDP-glucuronic acid, and quantitatively analyzing methanol and UDP-glucuronic acid through high-efficiency gas phase, wherein the generated amount of methanol and UDP-glucuronic acid generated by the reaction is calculated, and the enzyme activity ratio (relative activity%) is the ratio of the generated amount of methanol and UDP-glucuronic acid generated by the reaction after treatment to the generated amount of methanol and UDP-glucuronic acid generated by free enzyme catalysis. As can be derived from a number of experimental analyses, FDH/F ald DH/ADH/UGThe DH@MOF coated tetrazymes have good protection effect on enzymes compared with free enzymes, and FDH/F ald DH/ADH/UGDH@MAF-7 gave the best comparison of enzyme protection, as shown in FIG. 8.
Claims (8)
1. A metal organic framework multienzyme composite catalytic material comprises formate dehydrogenase, formaldehyde dehydrogenase, methanol dehydrogenase, UDP-glucose dehydrogenase and MOF material, and is characterized in that the composite catalytic material is provided with a medium cavitation capsule, the formate dehydrogenase, the formaldehyde dehydrogenase and the methanol dehydrogenase are wrapped in the capsule, and the UDP-glucose dehydrogenase is fixed in the MOF material; the MOF material is a Zn-based MOFs material: ZIF-8, ZIF-90 or MAF-7; specifically, the formate dehydrogenase, formaldehyde dehydrogenase and methanol dehydrogenase are immobilized through an etching template ZPF to obtain FDH/F ald DH/ADH@ZPF solids followed by FDH/F pairs using MOFs material ald Co-immobilization of DH/ADH@ZPF and UDP-glucose dehydrogenase to give (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF, and taking ZPF as a sacrificial template by utilizing the difference of the ZPF and the MOF to obtain the hollow metal-organic framework FDH/F ald DH/ADH/UGDH@MOF composite catalytic material.
2. The metal-organic framework multienzyme composite catalytic material of claim 1, characterized in that the etched template ZPF is a zeolitic pyrimidine framework material.
3. A method for preparing the metal-organic framework multienzyme composite catalytic material according to claim 1, which is characterized by comprising the following steps:
(1) Preparation of FDH/F ald DH/ADH@ZPF: FDH, F ald DH. ADH is put into Tris-HCl buffer solution, then 2-hydroxy-5 fluoropyrimidine is added, finally zinc nitrate is added, and the mixture is stirred for a period of time, centrifuged, washed and dried to obtain solid;
(2) Preparation (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF: the obtained FDH/F ald DH/ADH@ZPF and UGDH are added into Tris-HCl buffer solution together, then organic ligand is added, zinc nitrate is added after stirring evenly, and stirring reaction is carried out for a period of timeThen, centrifuging, washing and drying to obtain a solid;
(3) Preparation of FDH/F by etching ZPF ald DH/ADH/UGDH@MOF: the prepared (FDH/F) ald DH/ADH@ZPF)/UGDH@MOF powder is added into an acidic buffer solution to be stirred, and after a period of time, the mixture is centrifuged, washed and dried to obtain a solid.
4. The method for preparing a metal-organic framework multienzyme composite catalytic material according to claim 3, characterized in that the FDH and F in step (1) ald DH. ADH concentration of 0.3-0.5 mg/mL, tris-HCl buffer pH of 7-8, concentration of 0.1-0.3M, 2-hydroxy-5 fluoropyrimidine concentration of 0.2-0.4M, zinc nitrate concentration of 0.05-0.1M, reaction at 35-45 ℃ and 200-400 rpm for 8-12 h, and drying temperature of 35-40 ℃; the concentration of UGDH in the step (2) is 0.2-0.5mg/mL, the pH of Tris-HCl buffer solution is 7-8, the concentration is 0.1-0.3M, the organic ligand is one of 2-formaldehyde imidazole, 2-methyl imidazole and 3-methyl-1,2,4-triazole, zinc nitrate is 30-50 mM, the reaction is carried out at 35-45 ℃ for 12-24 h at 200-400 rpm, and the drying temperature is 35-40 ℃; the acidic buffer solution in the step (3) is PB buffer solution, the pH is 3.5-5.5, the concentration is 50-70 mM, the etching time is 0.5-1 h, and the drying temperature is 35-40 ℃.
5. Use of the metal-organic framework multienzyme composite catalytic material of claim 1 for carbon neutralization.
6. Use of the metal-organic framework multienzyme composite catalytic material of claim 1 in the production of glycosaminoglycans.
7. A method of using the metal-organic framework multienzyme composite catalytic material of claim 1, comprising the steps of:
(1) CO is processed by 2 Continuously introducing pure water and preparing NADH solution;
(2) To be saturated with CO 2 After the solution preparation is completed, the freshly prepared NADH solution is added with CO 2 Adding UDP-glucose and FDH/F into the solution ald DH/ADH/UGDH@MOF composite catalytic material is uniformly stirred after being sealed and subjected to oscillation reaction;
(3) Centrifuging the reaction solution in the step (2), collecting the composite catalyst after centrifugation, washing for recycling, taking the supernatant of the centrifugate, and detecting the generated methanol and UDP-glucuronic acid by a gas chromatography method.
8. The method of claim 7, wherein the CO of step (1) 2 The aeration time is 30-50 min, and the NADH concentration is 30-50 mM; step (2) the F ald The adding amount of the DH/ADH/UGDH@MOF composite catalytic material is 5% -10% of the mass of the substrate; the oscillating reaction is a water bath oscillating reaction at 20-50 ℃, the rotating speed is 100-300 rpm, and the reaction time is 10-24 hours; the centrifugation condition in the step (3) is 8,000-12,000 rpm for 4-6 min.
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| WO2015097020A1 (en) * | 2013-12-24 | 2015-07-02 | Stichting Dienst Landbouwkundig Onderzoek | Process for reducing carbon dioxide to methanol |
| CN111269908A (en) * | 2020-02-28 | 2020-06-12 | 南开大学 | Preparation of large-space bioreactor based on covalent organic framework material capsule |
| CN111320760A (en) * | 2020-02-28 | 2020-06-23 | 南开大学 | Porous framework material, enzyme preparation containing porous framework material, preparation method and application |
| CN111876406A (en) * | 2020-06-18 | 2020-11-03 | 南京师范大学 | Magnetic nanoparticle-lipase-metal organic framework composite catalytic material and preparation method and application thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015097020A1 (en) * | 2013-12-24 | 2015-07-02 | Stichting Dienst Landbouwkundig Onderzoek | Process for reducing carbon dioxide to methanol |
| CN111269908A (en) * | 2020-02-28 | 2020-06-12 | 南开大学 | Preparation of large-space bioreactor based on covalent organic framework material capsule |
| CN111320760A (en) * | 2020-02-28 | 2020-06-23 | 南开大学 | Porous framework material, enzyme preparation containing porous framework material, preparation method and application |
| CN111876406A (en) * | 2020-06-18 | 2020-11-03 | 南京师范大学 | Magnetic nanoparticle-lipase-metal organic framework composite catalytic material and preparation method and application thereof |
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