Nanocomposites of fullerenes / photosensitizers, methods of making and use thereof

Abstract

The application discloses a fullerene / photoredox nanocomposite, a preparation method and application thereof; wherein the fullerene / photoredox nanocomposite comprises a fullerene / photoredox donor-acceptor structure composite material and a cocatalyst on the fullerene / photoredox donor-acceptor structure composite material; the preparation method comprises the following steps: assembling fullerene and a photoredox to prepare the fullerene / photoredox donor-acceptor structure composite material; and depositing a cocatalyst Pt by an in-situ photodeposition method to prepare the fullerene / photoredox nanocomposite; and the application of the fullerene / photoredox nanocomposite in oxidizing NADH by photo-generated holes. The fullerene / photoredox nanocomposite increases the separation efficiency of electrons and holes, widens the absorption range of a spectrum, increases the utilization efficiency of light, realizes rapid photocatalytic hole oxidation of NADH by using the high-efficiency charge separation of the nanocomposite, and obtains NAD + with enzymatic activity and clean energy H2 as the only by-product.

Description

Technical Field

[0001] This invention relates to the field of nanocomposite materials technology, specifically to a fullerene / photosensitizer nanocomposite material, its preparation method, and its application. Background Technology

[0002] Enzyme-catalyzed reactions have advantages such as mild reaction conditions, fast reaction rates, and high product specificity. More than 90% of enzyme-catalyzed reactions involve the nicotinamide cofactor (NAD(H)), but it is very expensive. Various methods are used to obtain NAD. + This is a direction that researchers are focusing on.

[0003] Currently, methods for catalytic NADH oxidation, including enzyme catalysis, electrocatalysis, and organometallic catalysis, suffer from problems such as expensive and cumbersome enzyme expression, enzyme inactivation due to metal ions, and complex downstream reactions. Among these, photocatalysis is the most widely used method, with some organic photocatalysts, quantum dots (QDs), TiO2, and carbon-based nanomaterials successfully applied to NADH oxidation using photogenerated holes. However, organic dyes are easily deactivated by photobleaching and lack cycling stability. QDs are typically based on toxic heavy metals such as cadmium, which is unfriendly to biological systems. TiO2 has a band gap of 3.2 eV and a narrow spectral range, with absorption wavelengths mainly concentrated in the ultraviolet region (λ < 387 nm). Since only about 3% of sunlight reaches the Earth's surface, the visible light portion of solar energy cannot be utilized, resulting in very low solar energy utilization efficiency. Furthermore, the high recombination rate of photogenerated carriers leads to low quantum efficiency. These drawbacks of photocatalysts hinder their applicability. Summary of the Invention

[0004] To address the aforementioned problems, this invention provides a fullerene / photosensitizer nanocomposite material, its preparation method, and its applications. The donor-acceptor (DA) structure of the fullerene / photosensitizer nanocomposite material broadens the spectral absorption range and increases light utilization efficiency. Furthermore, the large dipole moment of the donor-acceptor (DA) structure of the fullerene / photosensitizer nanocomposite material increases the electron-hole separation efficiency, contributing to a significant improvement in photocatalytic performance and thus enhancing the efficiency of NADH oxidation. This enables the rapid and efficient production of enzymatically active NAD+. + It promotes the catalytic reactions of various enzymes.

[0005] As a first aspect, the present invention provides a fullerene / photosensitizer nanocomposite material, comprising a fullerene / photosensitizer donor-acceptor structural composite material and a cocatalyst deposited on the fullerene / photosensitizer donor-acceptor structural composite material, wherein the fullerene / photosensitizer donor-acceptor structural composite material is a fullerene and a photosensitizer assembled by non-covalent bonding or covalent bonding; the cocatalyst is Pt, wherein the mass fraction of the cocatalyst Pt is 1% to 13% of the fullerene / photosensitizer donor-acceptor structural composite material, preferably 5% to 10%.

[0006] In a second aspect, the present invention provides a method for preparing the above-mentioned nanocomposite material, comprising: preparing the above-mentioned fullerene / photosensitizer donor-acceptor composite material by non-covalent assembly or covalent assembly of fullerene and photosensitizer; depositing a co-catalyst Pt on the fullerene / photosensitizer donor-acceptor composite material by in-situ photodeposition to prepare a fullerene / photosensitizer nanocomposite material; wherein the above-mentioned non-covalent assembly adopts a method of directly assembling fullerene and photosensitizer; the above-mentioned covalent assembly adopts a method of first subjecting fullerene and photosensitizer to an addition reaction and then assembling them; preferably, the above-mentioned assembly method includes any one of the following: liquid-liquid interface method, solvent evaporation method, coprecipitation method, chemical vapor deposition method, surface adsorption method, or molecular template method.

[0007] As a third aspect, the present invention provides an application of the above-mentioned nanocomposite material in the photogenerated hole oxidation of NADH.

[0008] As a fourth aspect, the present invention provides the above-mentioned nanocomposite material for detecting NAD. + Applications in the presence or absence of enzyme activity, including: using NAD+ + Enzymes and substrates acting as cofactors are added to NAD+-containing compounds. + The test was conducted in a solution containing NAD; wherein, the above-mentioned NAD... + The cofactor enzyme is selected from one or more of glucose-6-phosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, α-ketoglutarate dehydrogenase, enolketoate dehydrogenase, or formate dehydrogenase; preferably, the above-mentioned enzyme is NAD+. + The cofactor enzyme is selected from alcohol dehydrogenase and / or formate dehydrogenase; wherein, for alcohol dehydrogenase, the general formula of the substrate is R-OH, where R is selected from -CH3, -C2H5, -C4H9, -C5H 11 One or more of -CH(CH3)2, -CH2CH(CH3)2, and -CH2C6H5; for the corresponding formate dehydrogenase, the substrate can be one or more of CH3OH, C2H5OH, and HCOOH.

[0009] As a fifth aspect, the present invention provides an application of the above-mentioned nanocomposite material in the simultaneous generation of hydrogen gas by photogenerated holes oxidizing NADH.

[0010] The fullerene / photosensitizer nanocomposite material provided by this invention increases the separation efficiency of electrons and holes, broadens the spectral absorption range, increases the utilization efficiency of light, and significantly improves the photocatalytic performance, thereby enhancing the efficiency of NADH oxidation and enabling the rapid and efficient production of NAD+ with enzymatic activity. + Fullerenes possess stable structures, high electronegativity and biocompatibility, and strong electron-accepting capabilities, making them excellent electron acceptors. Porphyrins and phthalocyanines, among other photosensitizers, have large π-conjugated aromatic faces and high molar extinction coefficients in the visible light range, making them typical electron donors. The donor-acceptor (DA) structure of the fullerene / photosensitizer donor-acceptor composite material exhibits a large dipole moment, increasing the electron-hole separation efficiency and significantly enhancing photocatalytic performance. Furthermore, during the photocatalytic oxidation of NADH, the fullerene / photosensitizer donor-acceptor composite material generates electron-hole pairs upon photoexcitation. NADH directly acts as an electron sacrificial agent, utilizing the holes to oxidize NADH into enzymatically active NAD. + Meanwhile, Pt in the co-catalyst utilizes electrons to produce hydrogen, generating hydrogen gas as a byproduct. Attached Figure Description

[0011] Figure 1 It is C 60 SEM image of the zinc donor-acceptor composite material of tetraphenylporphyrin;

[0012] Figure 2 It is C 60 / UV-Vis diffuse reflectance spectra of tetraphenylporphyrin zinc donor-acceptor composite material and tetraphenylporphyrin zinc;

[0013] Figure 3 It is C 60 / Tetraphenylporphyrin zinc donor-acceptor composite material, tetraphenylporphyrin zinc and C 60 XRD patterns;

[0014] Figure 4 It is C 60 XPS spectra of Zn in a tetraphenylporphyrin zinc donor-acceptor composite material;

[0015] Figure 5 It is C 60 XPS spectra of C in the zinc donor-acceptor composite material of tetraphenylporphyrin;

[0016] Figure 6 It is C 60 XPS spectra of N in the tetraphenylporphyrin zinc donor-acceptor composite material;

[0017] Figure 7 It is C60 Surface photovoltage spectrum of the tetraphenylporphyrin zinc donor-acceptor composite material;

[0018] Figure 8 It is NADH and NAD + The structural formula;

[0019] Figure 9 It is C 60 UV-Vis spectrum of NADH oxidation by tetraphenylporphyrin zinc nanocomposite material. Detailed Implementation

[0020] I. Definition

[0021] In this invention, unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, to better understand this invention, definitions and explanations of relevant terms are provided below.

[0022] For the purpose of clarity and concise description, features are described herein as part of some identical or separate embodiments; however, it will be understood that the scope of the invention may include some embodiments having a combination of all or some of the features described.

[0023] Based on the above description of the present invention, and in accordance with common technical knowledge and conventional methods in the field, various other modifications, substitutions, or alterations can be made without departing from the basic technical concept of the present invention.

[0024] Unless otherwise expressly stated, throughout the specification and claims, the term "comprising" or its variations such as "including" or "comprising" shall be understood to include the stated elements or components without excluding other elements or other components.

[0025] As used herein, the term "fullerene" is a series of spherical cluster molecules composed of an even number of carbon atoms, consisting of 12 five-membered rings and the rest six-membered rings; it is a cage-like structure composed of carbon atoms. Fullerenes include hollow fullerenes and metallofullerenes, wherein the hollow fullerene is a cage-like structure composed of a single carbon atom.

[0026] The term "metal fullerene" refers to a class of compounds with unique structures and properties formed by incorporating various atoms, ions, or atomic clusters within the carbon cage structure of fullerenes. These compounds are commonly referred to as endohedral fullerenes, typically denoted by M@C. 2n The form is represented as A₂C₂@C, where M represents a metallic element. For example, metallofullerenes (A₂C₂@C) 2b Or metallofullerene B3N@C 2bAt least one of the following, wherein: 39≤b≤44, and optionally 41 or 42; A is at least one of Sc, La, and Y; B is at least one of Sc, La, Y, Ho, Lu, and Er.

[0027] As used herein, the “fullerene / photosensitizer donor-acceptor composite material” refers to a composite material prepared from raw materials including fullerene and photosensitizer, wherein the fullerene and photosensitizer are stacked into fullerene derivatives through π-π conjugation to form an electron donor-acceptor complex, and the assembly structure of the composite material can be changed by adjusting the assembly conditions.

[0028] As used herein, the term "fullerene / photosensitizer nanocomposite material" refers to a nanocomposite material "with Pt co-catalyst deposited on a fullerene / photosensitizer donor-acceptor structure"; wherein, the Pt co-catalyst is attached to the fullerene / photosensitizer donor-acceptor structure composite material by in-situ photodeposition.

[0029] As used in this article, the term "photosensitizer" is also known as a sensitizer, photocrosslinker, or photosensitive agent. In photochemical reactions, it refers to substances that transfer light energy to reactants that are not sensitive to visible light to enhance or expand their photosensitivity.

[0030] The term "good solvent" refers to a solvent that has a strong ability to dissolve a solute; the term "poor solvent" refers to a solvent that has a weak ability to dissolve a solute.

[0031] As used herein, the term "liquid-liquid interface method" refers to the liquid-liquid interface deposition method, in which the poor solvent occupies the space of solute molecules in the good solvent at the interface between a good solvent and a poor solvent, and the solute precipitates out of the solution through assembly.

[0032] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following embodiments. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention.

[0033] II. Detailed Implementation

[0034] Constructing a system for the efficient oxidation of NADH using photogenerated holes requires maximizing solar energy utilization, suppressing electron-hole recombination, and promoting charge transport. An ideal photocatalyst should possess stronger visible light absorption, biocompatibility, stability, and a suitable band gap to oxidize NADH to obtain enzymatically active NAD+. +Constructing donor-acceptor (DA) organic supramolecular semiconductor materials is an effective strategy to improve photocatalytic activity. Their HOMO and LUMO energy levels are located on the electron donor and acceptor units, respectively, thus allowing control of the band structure by altering the donor or acceptor units. The DA structure not only enhances light absorption but also improves the efficient separation of charge carriers, thereby increasing photocatalytic activity.

[0035] Fullerenes are commonly used as electron acceptors due to their high electronegativity and biocompatibility. Furthermore, their stability greatly overcomes the instability of organic materials in the presence of water, oxygen, and light, allowing for the preparation of highly ordered, energy-level-matched fullerene composites that enhance photogenerated carrier separation efficiency. This makes them an ideal choice for studying the oxidation of NADH by photogenerated holes.

[0036] Based on the above-described inventive concept, the present invention provides a fullerene / photosensitizer nanocomposite material, comprising a fullerene / photosensitizer donor-acceptor structural composite material and a cocatalyst deposited on the fullerene / photosensitizer donor-acceptor structural composite material, wherein the fullerene / photosensitizer donor-acceptor structural composite material is a fullerene and a photosensitizer assembled by non-covalent bonding or covalent bonding; the cocatalyst is Pt, wherein the mass fraction of the cocatalyst Pt is 1% to 13% of the fullerene / photosensitizer donor-acceptor structural composite material, for example: 2%, 5%, 6.6%, 7.3%, 8.6%, 9.8%, 11.4%, 12.6%, preferably 5% to 10%, for example: 5.4%, 6.3%, 7.8%, 8.2%, 9.4%.

[0037] The fullerene / photosensitizer nanocomposite material provided in this invention increases the separation efficiency of electrons and holes, broadens the spectral absorption range, increases the utilization efficiency of light, and significantly improves the photocatalytic performance, thereby enhancing the efficiency of NADH oxidation and enabling the rapid and efficient production of NAD+ with enzymatic activity. + Fullerenes possess stable structures, high electronegativity and biocompatibility, and strong electron-accepting capabilities, making them excellent electron acceptors. Porphyrins and phthalocyanines, among other photosensitizers, have large π-conjugated aromatic faces and high molar extinction coefficients in the visible light range, making them typical electron donors. The donor-acceptor (DA) structure of the fullerene / photosensitizer donor-acceptor composite material exhibits a large dipole moment, increasing the electron-hole separation efficiency and significantly enhancing photocatalytic performance. Furthermore, during the photocatalytic oxidation of NADH, the fullerene / photosensitizer donor-acceptor composite material generates electron-hole pairs upon photoexcitation. NADH directly acts as an electron sacrificial agent, utilizing the holes to oxidize NADH into enzymatically active NAD. + Meanwhile, Pt in the co-catalyst utilizes electrons to produce hydrogen, generating hydrogen gas as a byproduct.

[0038] In some embodiments of the present invention, the above-mentioned fullerene is selected from hollow fullerene C. 2a Metallofullerene A2C2@C 2b Or metallofullerene B3N@C 2b At least one of the following: 30≤a≤50, 39≤b≤44, A is at least one of Sc, La, Y, B is at least one of Sc, La, Y, Ho, Lu, Er, C represents carbon, and N represents nitrogen.

[0039] In some embodiments of the present invention, the above-mentioned fullerene is selected from hollow fullerene C. 2a Where 30≤a≤60, preferably, a is selected from 30, 35, 38, 39, 41 or 42; more preferably, a is 30 or 35.

[0040] In some embodiments of the present invention, the fullerene is selected from metallofullerene A2C2@C. 2b Or metallofullerene B3N@C 2b Any one of the following, wherein 39≤b≤44, A is any one of Sc, La, Y; B is any one of Sc, La, Y, Ho, Lu, Er, preferably A and B are Sc.

[0041] In some embodiments of the present invention, the fullerene is selected from metallofullerene A2C2@C. 2b Or metal Fuller B3N@C 2b Any one of the following, where b is selected from 40, 41 or 42.

[0042] In some embodiments of the present invention, the metal fullerene is a metal Sc fullerene derivative, preferably, the metal fullerene is Sc3N@C. 80 .

[0043] In some embodiments of the present invention, the photosensitizer is selected from at least one of metalloporphyrin or its derivatives, titanium cyanide or its derivatives, boron fluoride complexed dipyrrometheneboron difluoride (BODIPY), 4,5-Dibromofluorescein (DBF), perylene dyes, thiophene dyes, ruthenium bipyridine, coumarin, chlorophyll (Chl), eosin Y, porphyrin, rose red, rhodamine B (RDB), and methylene blue (MB).

[0044] In some embodiments of the present invention, the metalloporphyrin or its derivative is selected from one or more of zinc porphyrin or its derivative, copper porphyrin or its derivative; preferably, the metalloporphyrin derivative is selected from copper phenyl porphyrin and / or zinc phenyl porphyrin; more preferably, the metalloporphyrin derivative is selected from tetraphenylporphyrin zinc and / or tetra-carboxyphenylporphyrin zinc; the titanium cyanide or its derivative is selected from one or more of titanium cyanide zinc or its derivative, titanium cyanide copper or its derivative; preferably, the titanium cyanide derivative is selected from one or more of octaoctoxyphthalocyanine copper, octaoctoxyphthalocyanine zinc, tetra-tert-butylphthalocyanine copper, and tetra-tert-butylphthalocyanine zinc; more preferably, the titanium cyanide derivative is selected from octaoctoxytitanium cyanide zinc; and the perylene dye is selected from perylene tetracarboxylic acid. Perylene diimide (PDI), N,N′-dioctyl-3,4,9,10-perylenedicarboximide (PTCDI), 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA), and perylenemonoimide (PMI) are selected from one or more of these dyes. More preferably, the perylene dyes are selected from perylene tetracarboxylcarboxylic dianhydride (PMI). The thiophene dye is selected from one or more of the following: 4,9-dihydro-s-indaceno[1,2-b:5,6-b']dithiophene (IDTT), dithieno[3′,2′:3,4;2″,3″:5,6]benzo[1,2-c][1,2,5]thiadiazole (DTBT), and dibenzothiophene (DBT). More preferably, the thiophene dye is selected from dithieno[3′,2′:3,4;2″,3″:5,6]benzo[1,2-c][1,2,5]thiadiazole (DTBT). The chlorophyll is selected from chlorophyll a (Chlorophyla, Chl a), chlorophyll b (Chlorophyl b, Chl b), and chlorophyll c (Chlorophyla, Chl a). Chlorophyllin c, Chl c) or one or more of their derivatives, more preferably, the chlorophyll is selected from chlorophyll a (Chlorophyl a, Chl a).

[0045] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is a hollow fullerene phenylporphyrin derivative selected from C 60 / metallic zinc phenylporphyrin, C 60 / metallic copper phenylporphyrin, C 70 / metallic zinc phenylporphyrin, C 70 One or more of the following: copper phenylporphyrin.

[0046] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is selected from metallofullerene A2C2@C 2b / metallic zinc phenylporphyrin or metallic fullerene B3N@C 2b / metallic zinc phenyl porphyrin, wherein A and B are preferably Sc, and b is selected from 40, for example: metal fullerene B3N@C 2b / metallic zinc phenyl porphyrin, wherein B is selected as Sc and b is selected from 40.

[0047] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is formed by assembling fullerene and titanium cyanide or its derivatives.

[0048] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is a hollow fullerene C0. 2a Metal phthalocyanine derivatives, selected from C 60 / Octyloxyphthalocyanine copper, C 60 / Zinc octyloxyphthalocyanine, C 70 / Octyloxyphthalocyanine copper, C 70 / Zinc octyloxyphthalocyanine, C 60 / Tetra-tert-butylphthalocyanine copper, C 70 / Tetrateryl phthalocyanine copper, C 60 / Tetratert-butylphthalocyanine zinc, C 70 One or more of the following: tetra-tert-butyl phthalocyanine zinc.

[0049] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is a metallofullerene B3N@C 2b / Metal phthalocyanine derivatives, selected from Sc3N@C 80 / Octaoctyloxyphthalocyanine copper, Sc3N@C 80 / Octaoctyloxyphthalocyanine zinc, Sc3N@C 80 / Fourth-degree butylphthalocyanine copper, Sc3N@C 80 One or more of the following: tetra-tert-butyl phthalocyanine zinc.

[0050] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is a hollow fullerene C0. 2a / Perylene dye derivatives, selected from C 60 / Perylene diimide (PDI), C 60 / N,N′-Dioctyl-3,4,9,10-Perylenedicarboximide (PTCDI), C 60 / 3,4,9,10-Tetracarboxylic acid dianhydride perylene (PTCDA), C 60 / Perylene imide (PMI), C 70 / Perylene diimide (PDI), C 70 / N,N′-Dioctyl-3,4,9,10-Perylenedicarboximide (PTCDI), C 70 / 3,4,9,10-Tetracarboxylic acid dianhydride perylene (PTCDA), C 70 One or more of perylene monoimide (PMI).

[0051] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is a hollow fullerene C0. 2a / Thiophene dye derivatives, selected from C 60 / Indexed dithiophene (IDTT), C 60 / Dithiophenebenzothiadiazole (DTBT), C 60 / Dibenzothiophene (DBT), C 70 / Indexed dithiophene (IDTT), C 70 / Dithiophenebenzothiadiazole (DTBT), C 70 One or more of dibenzothiophene (DBT).

[0052] In some embodiments of the present invention, the donor-acceptor structure in the above-mentioned fullerene / photosensitizer nanocomposite material is a hollow fullerene C0. 2a / Chlorophyll derivatives, selected from C 60 Chlorophyll a (Chl a), C 60 Chlorophyll b (Chl b), C 60 Chlorophyll C (Chl c), C 70 Chlorophyll a (Chl a), C 70 Chlorophyll b (Chl b), C 70 One or more of chlorophyll c (Chl c).

[0053] According to some embodiments of the present invention, a method for preparing the above-mentioned nanocomposite material is provided, comprising: preparing the above-mentioned fullerene / photosensitizer donor-acceptor composite material by non-covalent assembly or covalent assembly of fullerene and photosensitizer; depositing a co-catalyst Pt on the fullerene / photosensitizer donor-acceptor composite material by in-situ photodeposition to prepare a fullerene / photosensitizer nanocomposite material; wherein, the above-mentioned non-covalent assembly adopts a method of directly assembling fullerene and photosensitizer; the above-mentioned covalent assembly adopts a method of first performing an addition reaction of fullerene and photosensitizer and then assembling them; preferably, the above-mentioned assembly method includes any one of liquid-liquid interface method, solvent evaporation method, coprecipitation method, chemical vapor deposition method, surface adsorption method or molecular template method, more preferably, liquid-liquid interface method, solvent evaporation method or covalent bonding method. On the one hand, fullerenes and photosensitizers can form compounds through covalent bonds, and these compounds can form assemblies through any one of the following methods: liquid-liquid interface method, solvent evaporation method, coprecipitation method, chemical vapor deposition method, surface adsorption method, or molecular template method. On the other hand, fullerenes and photosensitizers can also directly form assemblies through any one of the following methods: liquid-liquid interface method, solvent evaporation method, coprecipitation method, chemical vapor deposition method, surface adsorption method, or molecular template method.

[0054] In some embodiments of the present invention, "in-situ photodeposition" refers to the following: in a photocatalytic reaction, the Pt precursor chloroplatinic acid undergoes physical adsorption near the photocatalyst (e.g., the fullerene / photosensitizer nanocomposite material of the present invention), and subsequent photoreduction leads to in-situ photodeposition of Pt clusters on the surface layer of the photocatalyst. The specific steps of the "in-situ photodeposition" include: adding a certain volume of chloroplatinic acid solution to a uniformly dispersed PBS buffer containing the photocatalyst and NADH; purging the system with N2 to remove air; and irradiating the system with a light source containing visible light with λ > 400 nm to complete the Pt deposition.

[0055] In some embodiments of the present invention, a method for preparing a fullerene / photosensitizer nanocomposite material is provided, comprising the following steps: (1) preparing a non-covalently linked fullerene / photosensitizer donor-acceptor composite material using different assembly methods, or first performing an addition reaction between the fullerene and the photosensitizer, followed by molecular assembly, to prepare a fullerene / photosensitizer donor-acceptor composite material; (2) filtering the obtained fullerene / photosensitizer donor-acceptor composite material and then vacuum drying it at 60°C for 12 h; (3) depositing the co-catalyst Pt onto the fullerene / photosensitizer donor-acceptor composite material in solution using in-situ photodeposition. Wherein, the bonding mode between the fullerene and the photosensitizer in the fullerene / photosensitizer donor-acceptor composite material obtained by different assembly methods is covalently linked or non-covalently linked.

[0056] In some embodiments of the present invention, fullerenes and photosensitizers are stacked and assembled through π-π conjugation to form an electron donor-acceptor complex, thereby forming a composite material with a fullerene / photosensitizer donor-acceptor structure.

[0057] In some embodiments of the present invention, fullerene and photosensitizer are covalently bonded and then formed into an electron donor-acceptor complex by solvent evaporation, thereby forming a composite material with a fullerene / photosensitizer donor-acceptor structure.

[0058] In some embodiments of the present invention, the preparation method of assembling fullerene and photosensitizer into a fullerene / photosensitizer donor-acceptor composite material includes any one of the following: liquid-liquid interface method, solvent evaporation method, co-precipitation method, chemical vapor deposition method, surface adsorption method, or molecular template method; wherein, the liquid-liquid interface method refers to the stacking of fullerene and photosensitizer through π-π conjugation to form a fullerene / photosensitizer donor-acceptor composite material.

[0059] In some embodiments of the present invention, a liquid-liquid interface method is used to stack the fullerene / photosensitizer nanocomposite material through π-π conjugation to form a donor-acceptor structure composite material. The specific preparation method includes the following steps: (1) Fullerene and photosensitizer are fully dissolved in a good solvent at a certain molar ratio; (2) A poor solvent is added to the solution prepared in step (1) at a certain volume ratio by dropwise addition or rapid mixing, allowed to stand for sedimentation for several hours, filtered to remove the solvent, and the resulting solid is collected to obtain the donor-acceptor structure material of the fullerene / photosensitizer nanocomposite material. The molar ratio of fullerene to photosensitizer is 1:0.1 to 1:10, preferably 1:0.5 to 1:2, and more preferably 1:1. The concentration of fullerene in the good solvent is 0.1 mg / mL to 5 mg / mL, preferably 0.5 mg / mL to 2 mg / mL, and more preferably 1 mg / mL. The standing deposition time is 1 h to 12 h, preferably 3 h to 8 h. Preferably, the good solvent is selected from one or more of toluene, o-xylene, m-xylene, p-xylene, 1,3,5-trimethylbenzene, chlorobenzene, o-dichlorobenzene, and carbon disulfide. Preferably, the good solvent is selected from 1,3,5-trimethylbenzene. Preferably, the poor solvent is selected from one or more of methanol, ethanol, isopropanol, n-propanol, n-butanol, and ethylene glycol. Preferably, the poor solvent is selected from isopropanol. The volume ratio is the volume ratio of good solvent to poor solvent. Specifically, the volume ratio of good solvent to poor solvent is 1:1 to 1:20. Preferably, the volume ratio of good solvent to poor solvent is 1:1 to 1:6. More preferably, the volume ratio of good solvent to poor solvent is 1:2 or 1:3.

[0060] In some embodiments of the present invention, the covalent bonding assembly method of first reacting a fullerene with a photosensitizer via an addition reaction and then assembling the photosensitizer includes the following steps: in the presence of an amino acid, the fullerene and photosensitizer undergo an addition reaction, wherein the addition reaction can be any one of 1,3-dipolar cycloaddition, Binger reaction, [2+2] cycloaddition, [2+4] cycloaddition, and carbene addition, preferably the 1,3-dipolar cycloaddition; wherein the molar ratio of fullerene to photosensitizer can be 1:1 to 1:1.5; the assembly method is to perform molecular assembly of fullerene / photosensitizer using a solvent evaporation method to obtain a fullerene / photosensitizer donor-acceptor composite material, wherein the solvent evaporation method is a good solvent and a poor solvent evaporation method, wherein the volume ratio of good solvent to poor solvent is 1:0.1 to 1:20, preferably 1:3.

[0061] In some embodiments of the present invention, the addition reaction in the covalent bonding method for preparing the fullerene / photosensitizer-receptor composite material is a 1,3-dipolar cycloaddition reaction.

[0062] In some embodiments of the present invention, the molar ratio of fullerene to photosensitizer in the covalent bonding method of the fullerene / photosensitizer-receptor composite material can be selected as 1.5.

[0063] In some embodiments of the present invention, the volume ratio of good solvent to poor solvent in the covalent bonding method of the fullerene / photosensitizer acceptor composite material is 1:3.

[0064] In some embodiments of the present invention, the good solvent in the covalent bonding method of the fullerene / photosensitizer-receptor composite material is 1,3,5-trimethylbenzene; the bad solvent is isopropanol.

[0065] In some embodiments of the present invention, the fullerene / photosensitizer donor-acceptor composite material is prepared by fullerene and metalloporphyrin or their derivatives using a liquid-liquid interface method, and then assembled by a solvent evaporation method. The terms "liquid-liquid interface method" and "solvent evaporation method" are explained as follows: (1) Conceptually: The "liquid-liquid interface method" is a method of depositing material on an interface formed between two immiscible liquids by utilizing the interaction at the interface. When a good solvent comes into contact with a poor solvent, due to their incompatibility, material particles precipitate at the interface, forming and gradually depositing on the interface. As the reaction proceeds, the resulting material particles aggregate and form nanoparticles. The "solvent evaporation method" is a method of forming material by dissolving it in a solution and allowing the solvent in the solution to gradually evaporate. First, the desired material is dissolved in a good solvent to form a solution. Then, the solution is dropped onto the surface of a poor solvent. Over time, the solvent begins to evaporate, causing the material to gradually precipitate from the solution. (2) The differences between the "liquid-liquid interface method" and the "solvent evaporation method" are as follows: First, in terms of "basic principles": the "liquid-liquid interface method" relies on the difference in solubility between incompatible liquids to precipitate the assembly, while the "solvent evaporation method" mainly controls the formation of the material through the evaporation of the solvent. Second, in terms of "reaction environment": the "liquid-liquid interface method" requires an interface between two immiscible liquids, while the "solvent evaporation method" does not require a clear liquid-liquid interface, only an interface between the solution and the gas phase. Furthermore, in this invention, in the "liquid-liquid interface method," the good solvent is 1,3,5-trimethylbenzene, which is rapidly injected into the poor solvent; in the "solvent evaporation method," the good solvent is 1,3,5-trimethylbenzene, which is slowly added to the surface of the poor solvent. Specifically, when using non-covalent bonding assembly, the assembly is prepared using the "liquid-liquid interface method"; for covalent bonding assembly, the assembly is prepared using the "solvent evaporation method" after the addition reaction.

[0066] In some embodiments of the present invention, a method for preparing a fullerene / photosensitizer donor-acceptor composite material by assembling fullerene with porphyrin or its derivatives is provided. The fullerene / photosensitizer donor-acceptor composite material is obtained by covalent bonding of fullerene with porphyrin or its derivatives. The preparation method includes: (1) performing a 1,3-dipolar cycloaddition reaction of fullerene with porphyrin or its derivatives in the presence of amino acids; (2) performing molecular assembly of fullerene / porphyrin or its derivatives by solvent evaporation to obtain the fullerene / photosensitizer donor-acceptor composite material. The molar ratio of fullerene to photosensitizer can be selected as 1:1.5. The solvent evaporation method is a good solvent and a bad solvent evaporation method. The good solvent is 1,3,5-trimethylbenzene, and the bad solvent is isopropanol. The volume ratio of the good solvent to the bad solvent is 1:3.

[0067] According to some embodiments of the present invention, the present invention provides an application of the above-described nanocomposite material in the photogenerated hole oxidation of NADH.

[0068] In some embodiments of the present invention, the application of any of the above-described fullerene / porphyrin derivative nanocomposites in the photogenerated hole oxidation of NADH is provided.

[0069] In some embodiments of the present invention, the application of any of the above-described hollow fullerene / porphyrin derivative nanocomposites in the photogenerated hole oxidation of NADH is provided; wherein, the hollow fullerene / porphyrin derivative composite material is selected from C 60 / Phenyloporphyrin, C 60 / metallic zinc phenylporphyrin, C 60 / metallic copper phenylporphyrin, C 70 / metallic zinc phenylporphyrin, C 70 One or more of the following: copper phenylporphyrin.

[0070] In some embodiments of the present invention, the application of any of the above-described fullerene / titanium cyanine derivative nanocomposites in the photogenerated hole oxidation of NADH is provided; wherein, the fullerene / titanium cyanine composite material is selected from C 60 / Octyloxyphthalocyanine copper, C 60 / Zinc octyloxyphthalocyanine, C 70 / Octyloxyphthalocyanine copper, C 70 Any one of the following: / octyloxyphosphocyanine zinc.

[0071] In some embodiments of the present invention, the application of any of the above-described fullerene / perylene dye derivative nanocomposites in the photogenerated hole oxidation of NADH is provided; wherein, the fullerene / perylene dye composite material is selected from C 60 / Perylene diimide (PDI), C 60 / N,N′-Dioctyl-3,4,9,10-Perylenedicarboximide (PTCDI), C 70 / Perylene diimide (PDI), C 70 Any of / N,N′-dioctyl-3,4,9,10-perylenedicarboximide (PTCDI).

[0072] In some embodiments of the present invention, the application of any of the above-described fullerene / thiophene dye derivative nanocomposites in the photogenerated hole oxidation of NADH is provided; wherein, the fullerene / thiophene dye composite material is selected from C 60 / Dithiophenebenzothiadiazole (DTBT), C 70 / Indexed dithiophene (IDTT), C 70 / Dithiophenebenzothiadiazole (DTBT), C 70Any of the dibenzothiophene (DBT) compounds.

[0073] In some embodiments of the present invention, the application of any of the above-described fullerene / chlorophyll derivative nanocomposites in the oxidation of NADH by photogenerated holes is provided; wherein, the fullerene / chlorophyll composite material is selected from C 60 Chlorophyll a (Chla), C 60 Chlorophyll b (Chl b), C 70 Chlorophyll b (Chl b), C 70 / Any of chlorophyll c (Chl c).

[0074] In some embodiments of the present invention, a method for using any of the above-mentioned nanocomposite materials for photogenerated hole oxidation of NADH is provided, comprising the following steps: adding a fullerene / photosensitizer acceptor composite material to a PBS buffer solution containing nicotinamide cofactor NADH, adding chloroplatinic acid solution, ultrasonically dispersing the mixture evenly, purging the air from the system with N2, irradiating the system with a light source containing visible light with λ > 400 nm, preferably λ being 400 nm to 800 nm, and detecting the NADH content by spectrophotometry at intervals; wherein, the fullerene / photosensitizer acceptor composite material is C 60 / Phenyloporphyrin, C 60 / metallic zinc phenylporphyrin, C 60 / metallic copper phenylporphyrin, C 70 / metallic zinc phenylporphyrin, C 70 / metallic copper phenylporphyrin, C 60 / Octyloxyphthalocyanine copper, C 60 / Zinc octyloxyphthalocyanine, C 70 / Octyloxyphthalocyanine copper, C 70 / Zinc octyloxyphthalocyanine, C 60 / Boron fluoride complexed dipyrrolemethyl guanylate fluorescent dye (BODIPY), C 60 / Perylene diimide (PDI), C 60 Chlorophyll a (Chl a), C 60 / 3,4,9,10-Tetracarboxylic acid dianhydride perylene (PTCDA), C 60 / Perylene imide (PMI), C 70 / Perylene diimide (PDI), C 70 / N,N′-Dioctyl-3,4,9,10-Perylenedicarboximide (PTCDI), C 70 / 3,4,9,10-Tetracarboxylic acid dianhydride perylene (PTCDA), C 70 / Perylene imide (PMI), C 60 / Indexed dithiophene (IDTT), C 60 / Dithiophenebenzothiadiazole (DTBT), C 60 / Dibenzothiophene (DBT), C 70 / Indexed dithiophene (IDTT), C 70 / Dithiophenebenzothiadiazole (DTBT), C 70 / Dibenzothiophene (DBT), C 60 Chlorophyll b (Chl b), C 60 Chlorophyll C (Chl c), C 70 Chlorophyll a (Chl a), C 70 Chlorophyll b (Chl b), C 70 One or more of chlorophyll c (Chl c); wherein the mass of platinum in the chloroplatinic acid solution is 1% to 13% of the mass of the fullerene / photosensitizer donor-receptor composite material, preferably, the concentration of the chloroplatinic acid solution is 0.1 mg / mL to 5 mg / mL, more preferably 0.5 mg / mL to 1.5 mg / mL, more preferably 0.8 mg / mL to 1.2 mg / mL, for example 0.9 mg / mL, 1.0 mg / mL or 1.1 mg / mL; preferably, the time for purging the air from the system with N2 is 20 min to 40 min, more preferably 25 min to 35 min, for example 28 min, 29 min, 30 min, 31 min or 32 min.

[0075] In some embodiments of the present invention, a method is provided for using any of the above-mentioned fullerene / photosensitizer donor-acceptor composite materials for photogenerated hole oxidation of NADH, comprising the following steps: adding 5 mg to 15 mg of photocatalyst, optionally 10 mg, to 55 mL of PBS buffer solution containing 1 mM NADH, then adding 1 mL of chloroplatinic acid solution, ultrasonically dispersing evenly, purging the system with N2 for 30 min to remove air, reacting with visible light with λ > 400 nm for 5 h, and detecting the NADH content by spectrophotometry after 1 h.

[0076] According to some embodiments of the present invention, an application of the above-described nanocomposite material is provided in the simultaneous generation of hydrogen gas by photogenerated holes oxidizing NADH.

[0077] In some embodiments of the present invention, a method is provided for using the above-mentioned nanocomposite material for photogenerated hole oxidation of NADH and photogenerated electron production of H2, comprising the following steps: adding the fullerene / photosensitizer acceptor structure composite material to an aqueous solution containing 5 mM NADH, adding a certain amount of chloroplatinic acid solution, ultrasonically dispersing the mixture evenly, evacuating the air from the system, irradiating the system with a light source containing visible light with λ > 400 nm, and detecting the volume of H2 by GC at intervals.

[0078] In some embodiments of the present invention, a method is provided for using the above-mentioned nanocomposite material for photogenerated hole oxidation of NADH and photogenerated electron production of H2, comprising the following steps: 5 mg to 15 mg (optionally 10 mg) of fullerene / photosensitizer to acceptor structure composite material is added to 100 mL of aqueous solution containing 5 mM NADH, and then 1 mL of chloroplatinic acid solution with a concentration of 1 mg / mL is added. After ultrasonic dispersion, the air in the system is evacuated, and the reaction is carried out with visible light with λ > 400 nm for 12 h. The volume of H2 is detected by GC every 1 h.

[0079] In some embodiments of the present invention, in any of the above-described methods for oxidizing NADH with photogenerated holes and reducing H2 with photogenerated electrons, the volume of the chloroplatinic acid solution is 0.5 mL to 1 mL.

[0080] In some embodiments of the present invention, in any of the above-described methods for oxidizing NADH with photogenerated holes and reducing H2 with photogenerated electrons, the visible light wavelength (λ) is 400 nm to 800 nm.

[0081] In some embodiments of the present invention, in any of the above-described methods for oxidizing NADH with photogenerated holes and reducing H2 with photogenerated electrons, the illumination time is 5h to 12h, optionally 8h.

[0082] In some embodiments of the present invention, in any of the above-described methods for generating H2 by oxidizing NADH with photogenerated holes and reducing H2 with photogenerated electrons, the intensity of the light source is 300 mW / cm². 2 ~500mw / cm 2 Preferably, 300mw / cm 2 .

[0083] According to some embodiments of the present invention, the above-described nanocomposite material is provided for the detection of NAD. + Applications in the presence or absence of enzyme activity, including: using NAD+ + Enzymes and substrates acting as cofactors are added to NAD+-containing compounds. + The test was conducted in a solution containing NAD; + The cofactor enzyme is selected from one or more of glucose-6-phosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, α-ketoglutarate dehydrogenase, enolketoate dehydrogenase, or formate dehydrogenase; preferably, the above-mentioned enzyme is NAD+. + The cofactor enzyme is selected from alcohol dehydrogenase and / or formate dehydrogenase; wherein, for alcohol dehydrogenase, the general formula of the substrate is R-OH, where R is selected from -CH3, -C2H5, -C4H9, -C5H 11One or more of -CH(CH3)2, -CH2CH(CH3)2, and -CH2C6H5; for the corresponding formate dehydrogenase, the substrate can be one or more of CH3OH, C2H5OH, and HCOOH.

[0084] In some embodiments of the present invention, a method for detecting NAD is provided. + A method for the presence or absence of enzyme activity, comprising the step of applying any of the fullerene / photosensitizers described above to a composite material of a receptor structure.

[0085] In some embodiments of the present invention, a method is provided for detecting NAD obtained from any of the fullerene / photosensitizer nanocomposites as described above. + The method for determining whether enzyme activity exists includes the following steps: After the NADH measured by spectrophotometry in the example of photogenerated hole oxidation of NADH has been consumed, the light source is turned off, and NAD+ is added. + The enzyme and substrate acting as cofactors were added to the solution, and a certain amount of solution was taken at different time points to detect the content of enzyme-catalyzed products.

[0086] In some embodiments of the present invention, any of the above-described methods for detecting NAD... + The method for determining whether an enzyme has activity can be used, and the enzyme may be one or more of glucose-6-phosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, α-ketoglutarate dehydrogenase, enolketoate dehydrogenase, or formate dehydrogenase, with NAD as the most important factor. + The cofactor enzyme is selected from alcohol dehydrogenase and / or formate dehydrogenase; the activity of alcohol dehydrogenase is 100 U / mL to 500 U / mL, preferably 300 U / mL; the volume of alcohol dehydrogenase is 1 mL to 10 mL, preferably 5 mL; the volume fraction of ethanol is 10% to 80%, preferably 50%; the volume of ethanol can be selected from 15 mL to 60 mL, preferably 45 mL.

[0087] In some embodiments of the present invention, any of the above-described methods for detecting NAD... + The method for determining whether an enzyme has activity corresponds to alcohol dehydrogenase, with the general formula for the substrate being R-OH, where R can be -CH3, -C2H5, -C4H9, or -C5H. 11 One or more of -CH(CH3)2, -CH2CH(CH3)2, -CH2C6H5 or -CH2CH2OH, further optionally -CH3, -C2H5 or -CH2C6H5, preferably -C2H5.

[0088] In some embodiments of the present invention, NAD is used. +The cofactor enzyme is selected from one or more of glucose-6-phosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, α-ketoglutarate dehydrogenase, enolketoate dehydrogenase, or formate dehydrogenase; preferably, the above-mentioned enzyme is NAD+. + The cofactor enzyme is selected from alcohol dehydrogenase and / or formate dehydrogenase; wherein, for alcohol dehydrogenase, the general formula of the substrate is R-OH, where R is selected from -CH3, -C2H5, -C4H9, -C5H 11 One or more of -CH(CH3)2, -CH2CH(CH3)2, and -CH2C6H5; for the corresponding formate dehydrogenase, the substrate can be one or more of CH3OH, C2H5OH, and HCOOH.

[0089] In one specific embodiment of the present invention, any of the above-described methods for detecting NAD... + The method for determining whether enzyme activity exists includes the following steps: After the NADH measured by spectrophotometry in the example of photogenerated hole oxidation of NADH has been consumed, the light source is turned off, and NAD+ is added. + 5 mL of alcohol dehydrogenase (acting as a cofactor) and 45 mL of ethanol were added to the solution. A certain amount of solution was taken at different time points and subjected to HPLC to detect the content of enzyme catalytic products.

[0090] III. Examples

[0091] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0092] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0093] In the following embodiments, the naming rules for composite materials are as follows:

[0094] (1) The name of a composite material containing (I) indicates that it is obtained by an assembly method of “non-covalent bonding”; where “non-covalent bonding” refers to “the method of directly assembling fullerene and photosensitizer”.

[0095] (2) The name of a composite material containing (II) indicates that it is obtained by the assembly method of "covalent bonding assembly". "Covalent bonding assembly" refers to "the method of first reacting fullerene and photosensitizer by addition reaction and then assembling".

[0096] Example 1: C 60 Preparation and property characterization of tetraphenylporphyrin zinc (I) donor-receptor composite materials

[0097] 1.1C 60 Preparation of tetraphenylporphyrin zinc (I) donor-receptor composite material

[0098] C 60The zinc tetraphenylporphyrin was mixed in a 1:1 molar ratio with the good solvent 1,3,5-trimethylbenzene. Then, the poor solvent isopropanol was added to the prepared good solution in a 1:3 volume ratio. After standing for 6 hours, the solvent was removed by filtration, and the resulting solid was collected and dried under vacuum to obtain the composite material.

[0099] 1.2C 60 Characterization of the properties of the tetraphenylporphyrin zinc (I) receptor-structured composite material

[0100] Figure 1 The scanning electron microscope (SEM) image of the sample clearly shows C. 60 The tetraphenylporphyrin zinc donor-acceptor composite material is well-assembled, has a regular morphology, and a particle size of about 500 nm.

[0101] Figure 2 C 60 The UV-Vis diffuse reflectance spectra of the tetraphenylporphyrin zinc (I) donor-acceptor composite material and tetraphenylporphyrin zinc were obtained. The light absorption capacity of the samples was tested using a Shimadzu UV-2550 UV spectrophotometer. Measurements were performed at room temperature, using BaSO4 as a reference, and the measurement wavelength range was 200 nm–800 nm. As can be seen from the figure, C… 60 The material synthesized with the tetraphenylporphyrin zinc (I) donor-acceptor structure has a wider visible light absorption region compared to tetraphenylporphyrin zinc.

[0102] Figure 3 C 60 / Tetraphenylporphyrin zinc (I) donor-acceptor composite material, tetraphenylporphyrin zinc and C 60 The XRD pattern was obtained. During the test, the wavelength of the X-rays was λ = 0.154 nm, the operating voltage was 40 kV, the operating current was 20 mA, the scanning speed was 10° / min, and the 2θ scanning range was 10–40°. The figure shows that compared to the corresponding tetraphenylporphyrin zinc monomer, the (001) crystal plane shifts to 7.4°, and C… 60 The disappearance of the typical crystal plane (101) at 10.74° and its shift to 11.33°, along with the increased intensity of the diffraction peaks at 19–22°, confirms that fullerene and zinc porphyrin have achieved assembly. The original monomer's crystal structure has changed, leading to corresponding changes in the diffraction peaks and intensities.

[0103] Figure 4 It is C 60 XPS spectra of Zn in a tetraphenylporphyrin zinc (I) donor-acceptor composite material; Figure 5 It is C 60 XPS spectra of C in the tetraphenylporphyrin zinc (I) donor-acceptor composite material; Figure 6 It is C 60XPS spectra of N in the tetraphenylporphyrin zinc (I) donor-acceptor composite material; analyzed using a Thermo Scientific ESCALab 250Xi multi-functional photoelectron spectrometer, with monochromatic Al Kα X-rays as the excitation source, approximately 200 W power, and an analysis area of ​​500 μm. Figure 4 , Figure 5 , Figure 6 It can be seen that C 60 The tetraphenylporphyrin zinc (1) donor-acceptor composite material is mainly composed of Zn, C, and N elements, C 1s Of the 288.8 eV values, 3.15% belonged to C=N, 0.33% to CN, and 96.52% to CC / C=C. 1s The 399.49 eV value belongs to C4H4N, Zn 2p The 1021.25 eV value belongs to ZnC4H4N.

[0104] Figure 7 It is C 60 Surface photovoltage spectrum of the donor-acceptor composite material of tetraphenylporphyrin zinc (1), compared with tetraphenylporphyrin zinc and C 60 Compared to single-component materials, the stronger response indicates that more charge can be transferred to the material surface.

[0105] Example 2: C 60 Photogenerated hole oxidation of NADH in tetraphenylporphyrin zinc (I) nanocomposites

[0106] The C obtained in Example 1 60 / Tetraphenylporphyrin zinc (1) 10 mg of the receptor structure composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, and then 1 mL of chloroplatinic acid solution was added. After ultrasonic dispersion, the air in the system was removed by passing N2 for 30 min. The reaction was carried out with visible light with λ>400 nm for 5 h. The NADH content was detected by spectrophotometry after 1 h.

[0107] Figure 8 It is NADH and NAD + The two structures have characteristic absorption peaks at 340 nm and 260 nm, respectively. The content of NADH can be determined by the decrease in absorbance at 340 nm. Figure 9 Representing C 60 The tetraphenylporphyrin zinc (I) composite material can completely oxidize NADH in 5 hours to obtain NAD. + The conversion rate is as high as 98.67%.

[0108] Table 1: C 60Performance testing of NADH oxidation of tetraphenylporphyrin zinc (1) composite material

[0109]

[0110] Table 1 shows that C60, Pt, and zinc tetraphenylporphyrin alone cannot oxidize NADH under the same test conditions, while only C60 can oxidize NADH. 60 The tetraphenylporphyrin zinc (I) donor-acceptor composite material requires 18 hours to oxidize NADH without the co-catalyst Pt, indicating that C 60 When used alone, both zinc tetraphenylporphyrin and zinc tetraphenylporphyrin do not have catalytic properties. However, when they are assembled into a nanoscale composite material and then photodeposited with a co-catalyst Pt, the transport capacity of photogenerated charges is improved, the recombination of photogenerated electrons and holes is greatly reduced, the quantum efficiency is improved, and photogenerated electrons can reach the catalyst surface and react.

[0111] The following experiments focus on C prepared under different reaction conditions. 60 The catalytic NADH oxidation performance of the tetraphenylporphyrin zinc (I) acceptor-structured composite material sample was tested.

[0112] Deposition time experiment: 5 groups containing C 60 The solutions of zinc tetraphenylporphyrin and zinc tetraphenylporphyrin were prepared by rapidly mixing isopropanol into the prepared 1,3,5-trimethylbenzene solution at a volume ratio of 1:2 to obtain a suspension. The suspension was allowed to stand at room temperature for 1, 2, 4, 6 and 8 hours, filtered, and washed three times with isopropanol to remove residual solvent. The suspension was then dried to obtain a solid powder. The photocatalytic NADH oxidation performance was tested, and the results are shown in Table 2.

[0113] Table 2: C 60 Photocatalytic NADH oxidation performance of tetraphenylporphyrin zinc (I) donor-acceptor composite material at different deposition times was tested.

[0114] Deposition time (h) Oxidation time of NADH (h) 1 15.5 2 11.5 4 6 6 5 8 7.5

[0115] As can be seen from Table 2, the two molecules require a certain deposition time to assemble, and nanocomposites with good catalytic NADH oxidation performance can be obtained by deposition of 4-8 hours.

[0116] Experiment on the ratio of good solvent to bad solvent: 4 groups containing C 60The solution of zinc tetraphenylporphyrin was rapidly mixed with isopropanol in the above-prepared solution at volume ratios of 1,3,5-trimethylbenzene:isopropanol = 1:1, 1:2, 1:3, and 1:4 to obtain a suspension. The suspension was allowed to stand at room temperature for 6 hours, filtered, and washed three times with isopropanol to remove residual solvent. The suspension was then dried to obtain a solid powder. The photocatalytic NADH oxidation performance was tested, and the results are shown in Table 3.

[0117] As can be seen from Table 3, composite materials with catalytic NADH oxidation performance can be obtained at ratios of 1:2 to 1:3, with a good solvent: bad solvent ratio of 1:3 being the preferred condition.

[0118] Table 3: C 60 Photocatalytic NADH oxidation performance of tetraphenylporphyrin zinc (I) donor-acceptor composite material under different solvent ratios was tested.

[0119] Good solvent: bad solvent Oxidation time of NADH (h) 1∶1 7.5 1∶2 5.5 1∶3 5 1∶4 6.5

[0120] C 60 / Tetraphenylporphyrin zinc (I) donor-receptor composite material with different dosages: 5 groups with different dosages of C 60 The photocatalytic NADH oxidation performance of the zinc donor-acceptor composite material was tested, and the results are shown in Table 4.

[0121] As shown in Table 4, nanocomposites with good catalytic NADH oxidation performance can be obtained at dosages of 8–13 mg, with 10 mg catalyst being the preferred condition.

[0122] Table 4: C 60 Photocatalytic NADH oxidation performance of tetraphenylporphyrin zinc (I) donor-acceptor composite material with different dosages was tested.

[0123] Catalyst (mg) Oxidation time of NADH (h) 3 17.5 5 12.5 8 9 10 5 13 5.5

[0124] Example 3: C 70 Preparation of tetraphenylporphyrin zinc (I) nanocomposites and photogenerated hole oxidation of NADH

[0125] 3.1C 70 Preparation of tetraphenylporphyrin zinc (I) donor-receptor composite material

[0126] C 70 The zinc tetraphenylporphyrin was mixed in a 1:1 molar ratio in a good solvent, 1,3,5-trimethylbenzene. Then, the poor solvent, isopropanol, was added to the prepared good solution in a 1:3 volume ratio. After standing for 6 hours, the solvent was removed by filtration, and the resulting solid was collected and dried under vacuum to obtain the composite material.

[0127] 3.2C 70 Photogenerated hole oxidation of NADH in tetraphenylporphyrin zinc (I) nanocomposites

[0128] The obtained C 70 10 mg of a tetraphenylporphyrin zinc (I) receptor-receptor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the system was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. 70 The tetraphenylporphyrin zinc (I) composite material completely oxidized NADH in 5.5 h to obtain NAD. + The conversion rate is as high as 97.54%.

[0129] Example 4: C 60 Preparation of / BODIPY(I) nanocomposites and photogenerated hole oxidation of NADH

[0130] 4.1C 60 / BODIPY(I) Preparation of receptor structure composite materials

[0131] C with a molar ratio of 1:1 60 The BODIPY and BODIPY were respectively mixed in 20 mL of the good solvent m-xylene. Then, the poor solvent isopropanol was added to the prepared good solution in a rapid mixing manner at a volume ratio of 1:3. After standing and sedimentation for 6 hours, the solvent was removed by filtration, and the resulting solid was collected and dried under vacuum to obtain the composite material.

[0132] 4.2C 60 / BODIPY(I) nanocomposite material for photogenerated hole oxidation NADH

[0133] The obtained C 60 / BODIPY(I) 10 mg of the receptor-structured composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the system was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 5.7 h to obtain NAD+. + The conversion rate is as high as 96.31%.

[0134] Example 5: C 60 Preparation of PDI(I) nanocomposites and photogenerated hole oxidation of NADH

[0135] C was prepared under the same conditions as in Example 4. 60 The / PDI(I) receptor-receptor composite material was prepared by adding 10 mg of the composite material to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air from the system. The reaction was then carried out under visible light (λ > 400 nm) for 5 h. NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH within 6.7 h to obtain NAD+. + The conversion rate is as high as 97.33%.

[0136] Example 6: C 60 Preparation of / PTCDI(I) nanocomposites and photogenerated hole oxidation of NADH

[0137] 6.1C 60 Preparation of / PTCDI(I) receptor structure composite material

[0138] C with a molar ratio of 1:1 60 PTCDI and o-xylene were mixed separately in 20 mL of good solvent. Then, isopropanol, a poor solvent, was added to the prepared good solution in a 1:3 volume ratio. The mixture was allowed to stand for 6 hours to settle, the solvent was removed by filtration, and the resulting solid was collected and dried under vacuum to obtain the composite material.

[0139] 6.2C 60 / PTCDI(I) nanocomposite photogenerated hole oxidation NADH

[0140] The obtained C 60 10 mg of the / PTCDI(I) receptor-receptor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the system was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 6.3 h to obtain NAD+. + The conversion rate is as high as 96.54%.

[0141] Example 7: C 60 Preparation of / DBF(I) nanocomposites and photogenerated hole oxidation of NADH

[0142] C was prepared under the same conditions as in Example 6. 60 / DBF(I) acceptor-donor composite material was prepared by adding 10 mg of the composite material to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air. The reaction was then carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 7.3 h to obtain NAD+. + The conversion rate is as high as 94.98%.

[0143] Example 8: C 60 Preparation of / Chl a(I) nanocomposites and photogenerated hole oxidation of NADH

[0144] 8.1C 60 Preparation of / Chl a(I) receptor structure composite material

[0145] C 60 Chl a was mixed with 20 mL of good solvent 1,3,5-trimethylbenzene at a molar ratio of 1:1. Then, n-butanol, a poor solvent, was added to the prepared good solution in a rapid mixing manner at a volume ratio of 1:3. After standing and sedimentation for 6 hours, the solvent was removed by filtration, and the resulting solid was collected and dried under vacuum to obtain the composite material.

[0146] 8.2C 60 / Chl a(I) nanocomposite photogenerated hole oxidation NADH

[0147] The obtained C 60 / Chl a(I) 10 mg of the receptor-receptor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the system was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h. The composite material completely oxidized NADH in 7.8 h to obtain NAD. + The conversion rate is as high as 98.54%.

[0148] Example 9: C 60 Preparation of / MB(I) nanocomposites and photogenerated hole oxidation of NADH

[0149] 9.1C 60 / MB(I) preparation of receptor structural composite materials

[0150] C 60MB was mixed with 20 mL of good solvent o-xylene at a molar ratio of 1:1. Then, n-butanol, a poor solvent, was added to the prepared good solution at a volume ratio of 1:3. After standing and sedimentation for 6 hours, the solvent was removed by filtration, and the resulting solid was collected and dried under vacuum to obtain the composite material.

[0151] 9.2C 60 / MB(I) nanocomposite material for photogenerated hole oxidation of NADH

[0152] The obtained C 60 10 mg of the / MB(I) receptor-receptor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air from the system. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 6.7 h to obtain NAD+. + The conversion rate is as high as 96.69%.

[0153] Example 10: C 70 Preparation of / RDB(I) nanocomposites and photogenerated hole oxidation of NADH

[0154] 10.1C 70 / RDB(I) preparation of receptor structure composite material

[0155] C 70 RDB and m-xylene were mixed in 20 mL of good solvent at a molar ratio of 1:1. Then, n-butanol, a poor solvent, was added to the prepared good solution in a rapid mixing manner at a volume ratio of 1:3. After standing and sedimentation for 6 hours, the solvent was removed by filtration, and the resulting solid was collected and dried under vacuum to obtain the composite material.

[0156] 10.2C 70 / RDB(I) nanocomposite photogenerated hole oxidation NADH

[0157] The obtained C 70 10 mg of the RDB(I) receptor-receptor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 8.2 h to obtain NAD+. + The conversion rate is as high as 97.52%.

[0158] Example 11: C 70 Preparation of / Chl b(I) nanocomposites and photogenerated hole oxidation of NADH

[0159] C was prepared under the same conditions as in Example 3. 70 The / Chl b receptor-receptor composite material was used. 10 mg of the composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval, based on the decrease in absorbance. The composite material completely oxidized NADH within 7 h to obtain NAD+. + The conversion rate is as high as 96.54%.

[0160] Example 12: Sc3N@C 80 Preparation of / TPP(II) nanocomposites and photogenerated hole oxidation of NADH

[0161] 12.1Sc3N@C 80 Preparation of / TPP(II) receptor-based composite materials

[0162] First, phenylporphyrin (TPP) and excess Cu(OAc)₂·H₂O were added to a mixed solvent of dichloromethane and methanol, and the mixture was heated and stirred for 2 hours. Rotary evaporation yielded a deep purple solid (CuTPP). Then, 0.74 mmol of CuTPP was dissolved in 30 mL of chloroform and 5 mL of DMF. The solution was cooled to 0°C in an ice bath under nitrogen protection, and phosphorus oxychloride was slowly added. The mixture was stirred at room temperature for 1 hour, and then the reaction mixture was heated to 70°C and reacted for 12 hours. Afterward, the solvent was removed by evaporation, and 96% sulfuric acid was slowly added dropwise in an ice bath while stirring overnight at room temperature. The reactants were then poured into continuously stirred water, and the pH was adjusted to neutral or weakly alkaline with saturated KOH solution. The mixture was extracted twice with chloroform, and the organic phase was dried over anhydrous magnesium sulfate and then passed through a silica gel column to collect a light purple powder solid β-formyl-tetraphenylporphyrin (Formyl-H₂P).

[0163] Then Sc3N@C 80 The product was mixed with β-formyl-tetraphenylporphyrin (Formyl-H2P) and a small amount of N- and glycine and dissolved in toluene solution. The solution was heated to 120°C under Ar protection and stirred for 2 h. The resulting solution was separated by HPLC, the target product peak was collected, evaporated to the solvent, and characterized by mass spectrometry and nuclear magnetic resonance.

[0164] Finally, the synthesized Sc3N@C 80 / TPP(II) was dissolved in a 1,3,5-trimethylbenzene solution, and isopropanol solution was slowly added to the surface of the solution. The mixture was left to stand at room temperature for 6 hours. The resulting precipitate was filtered, washed several times with isopropanol and methanol, and dried under vacuum to obtain Sc3N@C 80 / TPP(II) is a receptor-structured composite material.

[0165] 12.2Sc3N@C 80 / TPP(II) nanocomposite photogenerated hole oxidation NADH

[0166] The obtained Sc3N@C 80 10 mg of the / TPP(II) receptor-receptor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5.4 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH within 5 h to obtain NAD+. + The conversion rate is as high as 98.54%.

[0167] Example 13: C 70 Preparation of / TPP(II) nanocomposites and photogenerated hole oxidation of NADH

[0168] C was prepared and characterized under the same conditions as in Example 12. 70 The / TPP(II) acceptor-donor composite material was prepared by adding 10 mg of the composite material to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air from the system. The reaction was then carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 5.2 h to obtain NAD+. + The conversion rate is as high as 98.31%.

[0169] Example 14: C 60 Preparation of / TPP(II) nanocomposites and photogenerated hole oxidation of NADH

[0170] C was prepared and characterized under the same conditions as in Example 12. 60The / TPP(II) acceptor-donor composite material was prepared by adding 10 mg of the composite material to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the system was purged with N2 for 30 min to remove air. The reaction was then carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH within 5 h to obtain NAD+. + The conversion rate is as high as 97.11%.

[0171] Example 15: C 70 Preparation of / DTBT(II) nanocomposites and photogenerated hole oxidation of NADH

[0172] 15.1C 70 Preparation of / DTBT(II) receptor-structured composite materials

[0173] First, let C 70 The synthesized C14 was dissolved in toluene along with DTBT-CHO and a small amount of N-ethylglycine. The solution was sonicated until fully dissolved, then heated to 120°C under an Ar atmosphere, stirred, and refluxed for 6 hours. The resulting solution was separated by HPLC, the target product peak was collected, evaporated to the solvent, and characterized by mass spectrometry and NMR. Finally, the synthesized C14 was... 70 / DTBT(II) was dissolved in a 1,3,5-trimethylbenzene solution, and three times the volume of isopropanol solution was slowly added to the surface of the solution. The mixture was left to stand at room temperature for 6 hours. The resulting precipitate was filtered and washed several times with isopropanol and methanol, and then dried under vacuum to obtain C. 70 / DTBT(II) composite material.

[0174] 15.2C 70 / DTBT(II) nanocomposite photogenerated hole oxidation NADH

[0175] The obtained C 70 10 mg of the / DTBT(II) receptor-receptor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the mixture was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 6.8 h to obtain NAD+. + The conversion rate is as high as 97.36%.

[0176] Example 16: C 60Preparation of / DTBT(II) nanocomposites and photogenerated hole oxidation of NADH

[0177] C was prepared and characterized under the same conditions as in Example 15. 60 / DTBT(II) acceptor-donor composite material: 10 mg of the prepared acceptor-donor composite material was added to 55 mL of PBS buffer solution containing 1 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the system was purged with N2 for 30 min to remove air. The reaction was carried out under visible light (λ > 400 nm) for 5 h. The NADH content was detected by spectrophotometry at 340 nm after 1 h interval. The composite material completely oxidized NADH in 6.3 h to obtain NAD+. + The conversion rate is as high as 97.49%.

[0178] Example 17: Photocatalytic reduction of hydrogen production from fullerene / photosensitizer nanocomposites

[0179] Hydrogen is the only byproduct of this system. The catalytic hydrogen production performance of the material indirectly reflects the effectiveness of electron-hole separation; the stronger the hydrogen production performance, the better the ability of photogenerated holes to oxidize NADH. 10 mg of the prepared fullerene / photosensitizer acceptor composite material was added to 100 mL of an aqueous solution containing 5 mM NADH, followed by 1 mL of chloroplatinic acid solution. After ultrasonic dispersion, the air in the system was evacuated, and the reactor was irradiated with visible light (λ > 400 nm) for 12 h to induce a photocatalytic hydrogen production reaction. The hydrogen yield was determined by gas chromatography using the integrated area, and the corresponding hydrogen production performance of the fullerene / photosensitizer nanocomposite material was obtained, as shown in Table 5. The reaction still exhibited high catalytic activity after 3 hours. After cyclic testing, the morphology and structure of the sample remained consistent with those before the reaction, thus indicating good stability.

[0180] The photocatalytic water splitting process for hydrogen production involves the generation of electron-hole pairs after the photocatalyst is excited by light. An additional electron sacrificial agent (such as ascorbic acid) is introduced to scavenge these holes, reducing electron-hole recombination, and then the co-catalyst utilizes electrons to produce hydrogen. This invention directly uses NADH as an electron sacrificial agent, utilizing holes to oxidize NADH into enzymatically active NAD+. + Simultaneously, the catalyst utilizes electrons to produce hydrogen; in addition, NAD +The target product is hydrogen, which is a clean and easily removed byproduct. Therefore, the fullerene / photosensitizer nanocomposite material of this invention utilizes abundant and clean green light energy to catalyze NADH without pollution. Simultaneously, the reaction produces only one byproduct (i.e., clean energy hydrogen), which is easily removed from the system and does not affect downstream reactions. The catalytic performance of the fullerene / photosensitizer nanocomposite material reflects its electron-hole separation effect; a higher yield of the byproduct hydrogen indirectly reflects the strong hole oxidation ability of the fullerene / photosensitizer nanocomposite material for NADH.

[0181] Table 5: Photocatalytic hydrogen production performance test of fullerene / photosensitizer nanocomposites

[0182]

[0183]

[0184] Example 18: Fullerene / photosensitizer nanocomposite material generates NAD with enzymatic activity +

[0185] After the NADH measured by spectrophotometry in the photogenerated hole oxidation NADH test of each composite material was consumed, the light source was turned off, and NAD+ was added. + Add 5 mL of alcohol dehydrogenase (as a cofactor) and 45 mL of ethanol to the solution. After half an hour, take 2 mL of the solution to detect the content of acetaldehyde, the enzyme catalytic product.

[0186] We also used different enzymes and corresponding substrates to verify NAD. + The enzyme activity was measured, and specific data are shown in Table 6. It can be seen that the product content is related to NAD+. + There is a stoichiometric relationship, which can be expressed as: substrate (excess) + NAD + →Product + NADH.

[0187] Table 6: Verification of NAD using different materials and enzymes + enzyme activity

[0188]

[0189]

[0190] This shows that in various ways of utilizing NAD + In different enzyme systems, NAD+ acts as a cofactor. + All of them have activity, that is: the above different enzymes verify NAD. + The activity of the fullerene / photosensitizer nanocomposite material of the present invention is to oxidize NADH by photogenerated holes generated after photoexcitation, thereby obtaining NAD with enzymatic activity. + .

[0191] Under visible light irradiation, the fullerene / photosensitizer nanocomposite material achieved efficient separation of photogenerated electrons and holes, improving the efficiency of NADH oxidation by photogenerated holes, and thus producing NAD+ with enzymatic activity. + .

[0192] This invention discloses a class of non-covalently linked or chemically bonded fullerene / photosensitizer donor-acceptor composite materials. The fullerene is structurally stable and has a strong electron-accepting ability, acting as an electron acceptor. Porphyrins and phthalocyanine photosensitizers possess large π-conjugated aromatic surfaces and high molar extinction coefficients in the visible light range, making them typical electron donors and effectively broadening the spectral absorption range of novel catalysts. By assembling fullerenes and photosensitizers into donor-acceptor composite materials through non-covalent linkage or chemical bonding, the assembly structure characteristics can be modified by controlling factors such as the types of fullerenes and photosensitizers, increasing the utilization rate of sunlight, forming rapid charge transfer and efficient charge separation states, thereby improving quantum yield and constructing a novel photocatalyst for the efficient oxidation of NADH using photogenerated holes.

[0193] This invention utilizes abundant and clean light energy for catalysis, resulting in no pollution. The reaction produces only one byproduct (hydrogen), making it a clean energy source that is easily removed from the system and does not affect downstream reactions. Furthermore, the preparation method of the fullerene / photosensitizer nanocomposite material is simple and readily available. Photocatalytic water splitting for hydrogen production involves the generation of electron-hole pairs after photocatalyst excitation. An additional electron sacrificial agent (such as ascorbic acid) is introduced to remove these holes, reducing electron-hole recombination, and hydrogen is produced at the co-catalyst site using electrons. In the photocatalytic oxidation of NADH, the fullerene / photosensitizer nanocomposite material of this invention directly uses NADH as an electron sacrificial agent, utilizing holes to oxidize NADH into enzymatically active NAD+. + Simultaneously, the co-catalyst utilizes electrons to produce hydrogen from Pt, of which NAD... + The target product is hydrogen, which is a clean and easily removable byproduct.

[0194] The fullerene / photosensitizer nanocomposite material of this invention is easy to operate, non-toxic, and safe. It can be carried out under mild reaction conditions, is simple and controllable, and highly practical. Furthermore, it produces only one byproduct, H2, during the catalytic oxidation of NADH, making it a clean energy source. Compared to traditional inorganic semiconductor materials and organic dye molecules, this composite material exhibits structural stability and high order, high cycle stability, convenient recovery, and multiple reuses. The preparation method of the fullerene / photosensitizer nanocomposite material of this invention is simple and highly practical, making it an ideal choice for studying the photogenerated hole oxidation of NADH. In addition, this invention has strong universality, utilizing photogenerated hole oxidation to obtain enzymatically active NAD3. +It is applicable to catalysis by different types of enzymes, and the resulting substrate is compatible with NAD. + A stoichiometric relationship exists. Furthermore, the nanocomposite material provided by this invention not only effectively broadens the spectral absorption range and increases light utilization efficiency, but also, by utilizing the large dipole moment of the donor-acceptor structure, increases the separation efficiency of electrons and holes, facilitating the rapid and efficient oxidation of NADH to NAD by photogenerated holes. + And able to utilize the generated NAD + It promotes the catalytic reactions of various enzymes, and the byproduct of this system is clean energy H2.

[0195] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms described, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. Application of a fullerene / photosensitizer nanocomposite material in the photogenerated hole oxidation of NADH; The nanocomposite material comprises a fullerene / photosensitizer donor-acceptor composite material and a cocatalyst deposited on the fullerene / photosensitizer donor-acceptor composite material, wherein... The fullerene / photosensitizer donor-acceptor composite material is a fullerene and a photosensitizer assembled by non-covalent bonding or covalent bonding. The co-catalyst is Pt, wherein the mass fraction of the co-catalyst Pt is 1% to 13% of the fullerene / photosensitizer donor-acceptor composite material; The fullerene is selected from hollow fullerene C. 2a Metallofullerene A2C2@C 2b Or metallofullerene B3N@C 2b At least one of the following: 30≤a≤50, 39≤b≤44, A is at least one of Sc, La, Y, B is at least one of Sc, La, Y, Ho, Lu, Er, C represents carbon, and N represents nitrogen. The photosensitizer is selected from at least one of the following: metalloporphyrin or its derivatives, metal phthalocyanine or its derivatives, boron fluoride complexed dipyrrole methyl sulfone fluorescent dyes, dibromofluorescein, perylene dyes, indahedron dithiophene, dithiophene benzothiadiazole, dibenzothiaphene, ruthenium bipyridine, coumarin, chlorophyll, eosin Y, porphyrin, rose red, rhodamine B, and methylene blue; The metalloporphyrin or its derivative is selected from one or more of zinc porphyrin or its derivative, and copper porphyrin or its derivative; The metal phthalocyanine or its derivative is selected from one or more of zinc phthalocyanine or its derivative, copper phthalocyanine or its derivative; The perylene dyes are selected from one or more of perylene diimide, N,N′-dioctyl-3,4,9,10-perylene dicarboxylate, 3,4,9,10-tetracarboxylic dianhydride perylene, and perylene monoimide; The chlorophyll is selected from one or more of chlorophyll a, chlorophyll b, chlorophyll c, or their derivatives.

2. A fullerene / photosensitizer nanocomposite material for NAD detection + Applications of enzyme activity; The nanocomposite material comprises a fullerene / photosensitizer donor-acceptor composite material and a cocatalyst deposited on the fullerene / photosensitizer donor-acceptor composite material, wherein... The fullerene / photosensitizer donor-acceptor composite material is a fullerene and a photosensitizer assembled by non-covalent bonding or covalent bonding. The co-catalyst is Pt, wherein the mass fraction of the co-catalyst Pt is 1% to 13% of the fullerene / photosensitizer donor-acceptor composite material; The fullerene is selected from hollow fullerene C. 2a Metallofullerene A2C2@C 2b Or metallofullerene B3N@C 2b At least one of the following: 30≤a≤50, 39≤b≤44, A is at least one of Sc, La, Y, B is at least one of Sc, La, Y, Ho, Lu, Er, C represents carbon, and N represents nitrogen. The photosensitizer is selected from at least one of the following: metalloporphyrin or its derivatives, metal phthalocyanine or its derivatives, boron fluoride complexed dipyrrole methyl sulfone fluorescent dyes, dibromofluorescein, perylene dyes, indahedron dithiophene, dithiophene benzothiadiazole, dibenzothiaphene, ruthenium bipyridine, coumarin, chlorophyll, eosin Y, porphyrin, rose red, rhodamine B, and methylene blue; The metalloporphyrin or its derivative is selected from one or more of zinc porphyrin or its derivative, and copper porphyrin or its derivative; The metal phthalocyanine or its derivative is selected from one or more of zinc phthalocyanine or its derivative, copper phthalocyanine or its derivative; The perylene dyes are selected from one or more of perylene diimide, N,N′-dioctyl-3,4,9,10-perylene dicarboxylate, 3,4,9,10-tetracarboxylic dianhydride perylene, and perylene monoimide; The chlorophyll is selected from one or more of chlorophyll a, chlorophyll b, chlorophyll c, or their derivatives.

3. The application according to claim 1, characterized in that, The nanocomposite material generates hydrogen gas while oxidizing NADH with photogenerated holes.

4. The application according to any one of claims 1 to 3, characterized in that, The mass fraction of the cocatalyst Pt is 5% to 10% of the fullerene / photosensitizer donor-acceptor composite material.

5. The application according to claim 1, characterized in that, The metalloporphyrin derivative is selected from copper phenylporphyrin and / or zinc phenylporphyrin.

6. The application according to claim 1, characterized in that, The metalloporphyrin derivative is selected from tetraphenylporphyrin zinc and / or tetra-carboxyphenylporphyrin zinc.

7. The application according to claim 1, characterized in that, The metal phthalocyanine derivative is selected from zinc octyloxyphthalocyanine.

8. The application according to claim 1, characterized in that, The perylene dyes are selected from perylene diimides.

9. The application according to claim 1, characterized in that, The chlorophyll is selected from chlorophyll a.

10. The application according to any one of claims 1 to 3, characterized in that, The preparation method of the nanocomposite material includes: The fullerene / photosensitizer donor-acceptor composite material was prepared by a method of non-covalent or covalent assembly of fullerenes and photosensitizers. Fullerene / photosensitizer nanocomposite material was prepared by depositing cocatalyst Pt on fullerene / photosensitizer donor-acceptor composite material by in-situ photodeposition. in, The non-covalent assembly is performed by directly assembling fullerene and photosensitizer; The covalent bonding assembly is a method in which fullerene and photosensitizer undergo an addition reaction before assembly.

11. The application according to claim 10, characterized in that, The assembly method includes any one of the following: liquid-liquid interface method, solvent evaporation method, coprecipitation method, chemical vapor deposition method, surface adsorption method, or molecular template method.

12. The application according to claim 11, characterized in that, The liquid-liquid interface method refers to the formation of a fullerene / photosensitizer acceptor composite material through π-π conjugation of fullerene and photosensitizer, and the steps include: Fullerene and photosensitizer are dissolved in a good solvent at a certain molar ratio to obtain a mixed solution; The molar ratio of fullerene to photosensitizer is 1:0.1 to 1:10, and the concentration of fullerene in a good solvent is 0.1 mg / mL to 5 mg / mL. The undesirable solvent is added to the mixed solution in a certain volume ratio by dropwise addition or rapid mixing. After standing for 1 to 12 hours, the solvent is removed by filtration, and the resulting solid is collected to obtain the donor-acceptor structure of the fullerene / photosensitizer nanocomposite material.

13. The application according to claim 12, characterized in that, The molar ratio of fullerene to photosensitizer is 1:0.5 to 1:

2.

14. The application according to claim 12, characterized in that, The molar ratio of fullerene to photosensitizer is 1:

1.

15. The application according to claim 12, characterized in that, The concentration of fullerene in a good solvent is 0.5 mg / mL to 2 mg / mL.

16. The application according to claim 12, characterized in that, The concentration of fullerene in a good solvent is 1 mg / mL.

17. The application according to claim 12, characterized in that, Allow to settle for 3 to 8 hours.

18. The application according to claim 12, characterized in that, The good solvent is selected from one or more of toluene, o-xylene, m-xylene, p-xylene, 1,3,5-trimethylbenzene, chlorobenzene, o-dichlorobenzene, and carbon disulfide.

19. The application according to claim 12, characterized in that, The good solvent is selected from 1,3,5-trimethylbenzene.

20. The application according to claim 12, characterized in that, The unsuitable solvent is selected from one or more of methanol, ethanol, isopropanol, n-propanol, n-butanol, and ethylene glycol.

21. The application according to claim 12, characterized in that, The unsuitable solvent is selected from isopropanol.

22. The application according to claim 12, characterized in that, The volume ratio of the good solvent to the bad solvent is 1:1 to 1:

20.

23. The application according to claim 12, characterized in that, The volume ratio of the good solvent to the bad solvent is 1:1 to 1:

6.

24. The application according to claim 12, characterized in that, The volume ratio of the good solvent to the bad solvent is 1:2 to 1:

3.

25. The application according to claim 10, characterized in that, The covalent bonding assembly method involves first undergoing an addition reaction between fullerene and a photosensitizer, followed by assembly. The addition reaction step includes: In the presence of amino acids, fullerenes undergo an addition reaction with photosensitizers; The addition reaction is any one of 1,3-dipolar cycloaddition, Binger reaction, [2+2] cycloaddition, [2+4] cycloaddition, and carbene addition; the molar ratio of fullerene to photosensitizer is 1:1 to 1:1.

5. The assembly method involves using a solvent evaporation method to perform molecular assembly of fullerene / photosensitizer to obtain a fullerene / photosensitizer donor-acceptor composite material. Among them, the solvent evaporation method is the evaporation method of good solvent and bad solvent, wherein the volume ratio of good solvent to bad solvent is 1:0.1 to 1:

20.

26. The application according to claim 25, characterized in that, The addition reaction is a 1,3-dipolar cycloaddition reaction.

27. The application according to claim 25, characterized in that, The volume ratio of good solvent to poor solvent is 1:

3.

28. The application according to claim 1, wherein, Methods for photogenerated holes in nanocomposites to oxidize NADH include: The fullerene / photosensitizer receptor composite material was added to a PBS buffer solution containing nicotinamide cofactor NADH, chloroplatinic acid solution was added, and the mixture was ultrasonically dispersed. After the air in the system was removed by passing N2 through the solution, the system was irradiated with a light source containing visible light with λ > 400 nm. The NADH content was detected by spectrophotometry at intervals. The fullerene / photosensitizer donor-acceptor composite material is C 60 / metallic zinc phenylporphyrin, C 60 / metallic copper phenylporphyrin, C 70 / metallic zinc phenylporphyrin, C 70 / metallic copper phenylporphyrin, C 60 / Octyloxyphthalocyanine copper, C 60 / Zinc octyloxyphthalocyanine, C 70 / Octyloxyphthalocyanine copper, C 70 / Zinc octyloxyphthalocyanine, C 60 / Boron fluoride complexed dipyrrolemethyl benzoate fluorescent dyes, C 60 / Perylene diimide, C 60 Chlorophyll a, C 60 / 3,4,9,10-Tetracarboxylic acid dianhydride perylene, C 60 / Perylene imide, C 70 / Perylene diimide, C 70 / N,N′-Dioctyl-3,4,9,10-Perylenedicarboximide, C 70 / 3,4,9,10-Tetracarboxylic acid dianhydride perylene, C 70 / Perylene imide, C 60 / Yinda Province and dithiophene and thiophene, C 60 / Dithiophenebenzothiadiazole, C 60 / Dibenzothiophene, C 70 / Yinda Province and dithiophene and thiophene, C 70 / Dithiophenebenzothiadiazole, C 70 / Dibenzothiophene, C 60 Chlorophyll b, C 60 Chlorophyll C, C 70 Chlorophyll a, C 70 Chlorophyll b, C 70 One or more of chlorophyll C.

29. The application according to claim 28, characterized in that, The mass of platinum in the chloroplatinic acid solution is 1% to 13% of the mass of the fullerene / photosensitizer donor-receptor composite material.

30. The application according to claim 28, characterized in that, The concentration of the chloroplatinic acid solution is 0.1 mg / mL to 5 mg / mL.

31. The application according to claim 28, characterized in that, The concentration of the chloroplatinic acid solution is 0.5 mg / mL to 1.5 mg / mL.

32. The application according to claim 28, characterized in that, The concentration of the chloroplatinic acid solution is 0.8 mg / mL to 1.2 mg / mL.

33. The application according to claim 28, characterized in that, The concentration of the chloroplatinic acid solution is 0.9 mg / mL, 1.0 mg / mL, or 1.1 mg / mL.

34. The application according to claim 28, characterized in that, The time to purge the air from the system with N2 is 20 to 40 minutes.

35. The application according to claim 28, characterized in that, The time to purge the air from the system with N2 is 25 to 35 minutes.

36. The application according to claim 28, characterized in that, The time for purging air from the N2 system is 28 min, 29 min, 30 min, 31 min, or 32 min.

37. The application according to claim 28, characterized in that, λ is between 400 nm and 800 nm, and the value of λ is not 400 nm.

38. The application according to claim 2, characterized in that, The method for detecting the presence or absence of NAD+ enzyme activity includes: using NAD+... + Enzymes and substrates acting as cofactors are added to NAD+-containing compounds. + The test was conducted in the solution; Among them, the NAD + The cofactor enzyme is selected from one or more of glucose-6-phosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, α-ketoglutarate dehydrogenase, enolketoate dehydrogenase, or formate dehydrogenase. For alcohol dehydrogenase, the general formula of the substrate is R-OH, where R is selected from -CH3, -C2H5, -C4H9, and -C5H. 11 One or more of -CH(CH3)2, -CH2CH(CH3)2, and -CH2C6H5; The corresponding formate dehydrogenase has a substrate of HCOOH.

39. The application according to claim 38, characterized in that, The NAD + The cofactor enzyme is selected from alcohol dehydrogenase and / or formate dehydrogenase.

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