JPWO2020069227A5

JPWO2020069227A5 -

Info

Publication number: JPWO2020069227A5
Application number: JP2021517462A
Authority: JP
Publication date: 2022-09-27

Claims

A magnetic macroporous powder comprising a thermoplastic polymer and magnetic microparticles, operable for additive manufacturing (AM) of shaped magnetic macroporous scaffolds for immobilizing self-assembled mesoporous aggregates of magnetic nanoparticles. is a magnetic macroporous powder.

2. The magnetic macroporous powder of claim 1, wherein the magnetic particles have a size of about 50-100 μm , about 10-50 μm, about 5-10 μm, about 10 μm, about 5 μm, less than 5 μm, or greater than 100 μm .

2. The magnetic macroporous powder of claim 1, having an average size of about 150[mu] m, about 75[mu]m or about 15[mu ]m.

Magnetic macroporous powder according to claim 1, wherein the magnetic particles have a concentration of 0-10 wt% , 10-50 wt% or 50-90 wt% .

Said thermoplastic polymer is polyvinyl alcohol (PVA), acrylic (PMMA), acrylonitrile butadiene styrene (ABS), polyamide including nylon 6 and nylon 12, polylactic acid (PLA), polybenzimidazole (PBI), polycarbonate (PC) , polyethersulfone (PES), polyoxymethylene (POM), polyetheretherketone (PEEK), polyetherimide (PEI), polyethylene (PE), polyphenylene oxide (PEO), polyphenylene sulfide (PPS), polypropylene (PP ), polystyrene (PS), polyvinyl chloride (PVC), polytetrafluoroethylene (PTFE), copolyesters, and chemically functionalized derivatives thereof. Magnetic macroporous powder.

The magnetic microparticles include magnetite (Fe ₃ O ₄ ), hematite (α-Fe ₂ O ₃ ), maghemite (γ-Fe ₂ O ₃ ), spinel ferrite, natural magnets, cobalt, nickel, rare earths, and magnetic 2. The magnetic macroporous powder of claim 1, comprising a magnetic material selected from the group consisting of composites.

7. The magnetic macroporous powder of claim 6, wherein said rare earth is neodymium, gadolinium, cisprosium, samarium-cobalt, or neodymium-iron-boron.

7. The magnetic macroporous powder of claim 6, wherein said magnetic composite comprises ceramic, ferrite, or alnico magnets.

A magnetic macro according to any preceding claim , wherein said thermoplastic polymer and said magnetic microparticles are chemically blended, thermally blended or physically blended. porous powder.

Magnetic macroporous powder according to any one of claims 1 to 9 , comprising macropores with a size of 0.5 to 200 µm.

Magnetic macroporous powder according to any one of claims 1 to 10 , further comprising cellulose fibres, cellulose nanofibres, glass fibres, or carbon fibres.

A magnetic macro according to any one of claims 1 to 11 , further comprising self-assembled mesoporous aggregates of magnetic nanoparticles and an enzyme magnetically immobilized within said mesopores or on their surface. porous powder.

A shaped magnetic macroporous scaffold comprising a magnetic macroporous powder according to any one of claims 1 to 12 , said powder being formed into said shape by three-dimensional (3D) printing.

14. The shaped magnetic macroporous scaffold of claim 13 , wherein said shape is cylinder, sphere, bead, strip, capsule, cube, square rod, pyramid, diamond, lattice, or irregular shape.

15. The shaped magnetic macroporous scaffold of claim 13 or 14 , further comprising self-assembled mesoporous aggregates of magnetic nanoparticles.

16. The method of claim 15 , wherein the self-assembled mesoporous aggregates of magnetic nanoparticles further comprise one or more enzymes magnetically immobilized within or on the surface of the mesopores of the magnetic nanoparticles. Shaped magnetic macroporous scaffolds.

the one or more enzymes are hydrolase, hydroxylase, hydrogen peroxide generating enzyme (HPP), nitralase, hydratase, dehydrogenase, transaminase, ketoreductase (KREDS) enereductase (EREDS), imine reductase (IREDS), catalase , dismutase, oxidase, dioxygenase, lipoxidase, oxidoreductase, peroxidase, laccase, synthetase, transferase, oxynitrilase, isomerase, glucosidase, kinase, lyase, sucrase, invertase, epimerase, and lipase. 17. Shaped magnetic macroporous scaffold according to item 16 .

said self-assembled mesoporous aggregates of magnetic nanoparticles comprising microsomes, wherein a first enzyme requiring a diffusible cofactor having a first enzymatic activity is contained within said microsomes and comprises a cofactor regeneration activity a second enzyme is magnetically entrapped within said mesopores and said cofactor is utilized in said first enzymatic activity; said first and second enzymes diffusively generate a diffusible substrate; 16. The shaped magnetic macroporous scaffold of claim 15 , wherein said magnetic nanoparticles are magnetically associated with said magnetic macroporous scaffold.

said self-assembled mesoporous aggregates of magnetic nanoparticles comprising a first enzyme requiring a diffusible cofactor having a first enzymatic activity; a second enzyme comprising a cofactor regeneration activity, said cofactor comprising: utilized in said first enzymatic activity; said first and second enzymes are magnetically entrapped within said mesopores formed by said aggregates of magnetic nanoparticles; 16. The shaped magnetic macroporous scaffold of claim 15 , wherein the enzyme functions by converting a diffusible substrate to a diffusible product.

20. The shaped magnetic macroporous scaffold of claim 19 , wherein said first enzyme is an oxidase.

21. The shaped magnetic macro of claim 20 , wherein said oxidase is a flavin-containing oxygenase; and wherein said composition further comprises a third enzyme having co-factor reductase activity co-localized with said first enzyme. Porous scaffolding.

21. The shaped magnetic macro of claim 20 , wherein said oxidizing enzyme is a P450 monooxygenase; and wherein said composition further comprises a third enzyme having co-factor reductase activity co-localized with said first enzyme. Porous scaffolding.

21. The shaped magnetic macroporous scaffold of claim 20 , wherein said P450 monooxygenase and said third enzyme are contained within a single protein.

24. The shaped magnetic macroporous scaffold of claim 23 , wherein said single protein comprises a bifunctional cytochrome P450/NADPH--P450 reductase.

24. The shaped magnetic macroporous scaffold of claim 23 , wherein said single protein has BM3 activity and has at least 90% sequence identity with SEQ ID NO:1.

The shaped magnetic macroporous scaffold according to any one of claims 13-25 , wherein said scaffold is formed into a shape suitable for a particular biocatalytic process.

A method of forming a shaped magnetic macroporous scaffold comprising the magnetic macroporous powder of any one of claims 13-26 , wherein the shaped magnetic macroporous scaffold is formed using a three-dimensional (3D) printer. A method comprising additive manufacturing (AM) and obtaining said shape from a 3D model.

28. The method of claim 27 , wherein said 3D model is an electronic file.

29. The method of claim 28 , wherein the electronic file is a Computer Aided Design (CAD) or Stereolithography (STL) file.

The method of any one of claims 27-29 , wherein the AM is Fused Filament Fabrication (FFF) or Selective Laser Sintering (SLS).

31. The method according to any one of claims 27 to 30 , wherein said macropores are formed by using a soluble drug selected from the group consisting of salts, sugars or small soluble polymers and removing said soluble drug with a solvent. described method.

A method of forming a device for catalyzing an enzymatic reaction comprising combining a shaped magnetic macroporous scaffold with self-assembled mesoporous aggregates of magnetic nanoparticles and an enzyme, wherein the enzyme binds magnetic method comprising the step of physically immobilizing.

13. A method of catalyzing a reaction between a plurality of substrates, comprising exposing the magnetic macroporous powder of claim 12 to said substrate under conditions for said enzyme to catalyze said reaction between said substrates. .

18. A method of catalyzing a reaction between a plurality of substrates, wherein the shaped magnetic macroporous scaffold of claim 16 or 17 is exposed to said substrate under conditions in which said enzyme catalyzes said reaction between said substrates. A method that includes steps.

35. A method according to claim 33 or 34 , wherein said reaction is used in the production of pharmaceutical products , food products, flavors, fragrances, sweeteners, pesticides, antimicrobials, toxins, detergents, fuel products or biochemicals .