KR101190291B1 - Method for Photochemical Regenerating Oxidoreductase Cofactor Using Inorganic photosensitizer - Google Patents

Abstract

The present invention relates to a photochemical regeneration method of an oxidoreductase cofactor using an inorganic photosensitizer, and more particularly, to an oxidation type oxidoreductase cofactor using an inorganic photosensitizer and a redox mediator. The present invention relates to a photochemical regeneration method of an oxidoreductase cofactor which is converted to a reduced oxidoreductase cofactor.
According to the present invention, it is possible to rapidly regenerate the oxidoreductase cofactor by only irradiating light in the visible light region, and can be very useful for increasing the efficiency of various biocatalytic reactions using the oxidoreductase enzyme.

Description

Photochemical Regeneration of Oxidoreductase Cofactor Using Inorganic Photosensitive Agents {Method for Photochemical Regenerating Oxidoreductase Cofactor Using Inorganic photosensitizer}

The present invention relates to a photochemical regeneration method of an oxidoreductase cofactor using an inorganic photosensitizer, and more particularly, any one selected from the group consisting of methyl viologen, ruthenium II complex, and rhodium III complex. In the photochemical regeneration method of an oxidoreductase cofactor, the oxidative oxidoreductase cofactor is converted into a reduced oxidoreductase cofactor by using the above-described redox mediator and an inorganic photosensitizer. It is about.

Catalytic reactions with enzymes play an important role in producing the fine chemicals that are industrially necessary. However, the chemical catalysts used up to now mostly produce by-products such as stereoisomers during the production process, and the purification process increases the production cost of the material considerably. Unlike such chemical catalysts, when using a biocatalyst using an enzyme, it is possible to produce only a specific material without by-products. In particular, redox enzymes such as glutamate dehydrogenase (GDH), alcohol dehydrogenase (ADH) and lactate dehydrogenase (LDH) are mainly used to produce high-quality fine chemicals such as chiral compounds and alchol, aldehydes and amino acids. high.

However, there is a restriction that cofactors are required to use oxidoreductase. In the biocatalyst reaction using oxidoreductase, the cofactor plays a very important role in carrying out the enzyme reaction because it acts as a reducing equivalent. However, cofactors, which have two forms of oxidation or reduction, can no longer be used for enzymatic reactions once they are converted to one state. Therefore, the same amount as the enzyme substrate should always be used, considering that the cofactor is quite expensive, there are many economic difficulties in using it in the actual industry.

Therefore, researches for regenerating and reusing cofactors have been conducted, and as a result, many regeneration methods have been developed. Representative methods include biochemical methods and electrochemical methods using a second enzyme. Among them, a biochemical method using a second enzyme has a low thermodynamic equilibrium caused by formation of by-products or competition between substrates. As a result, the overall reaction rate is reduced. Regeneration of electrochemical cofactors also has a problem of poor regeneration efficiency due to the slow electron transfer rate between the electrode and the oxidoreductase cofactor of the oxidized type. To solve this problem, a method of transferring electrons between an electrode and a cofactor using a homogeneous redox mediator has been developed.

Specifically, methyl viologen has been used for indirect electrochemical regeneration with Flavoenzyme (Ferredoxin reductase (FDR) or Lipoamide Dehydrogenase (LipDH)) as an electron transfer medium for NAD (P) H ( Dicosimo et al., J Org Chem 46: 4622, 1981), electron transfer of the ruthenium (II) complex (hexamethylbenzene-2,2'-bipyridinechloro) ruthenium (II) to reduction of ketones to alcohols (Ogo S et al ., Organometallics 21: 2964, 2002; Yaw Kai Yan et al . J Biol Inorg Chem 11: 483, 2006) and the rhodium complex (pentamethylcyclopentadienyl-2) , 2'-bipyridinechloro) rhodium (III): [Cp * Rh (bpy) H ₂ O] ²⁺ (hereinafter M _ox ) was used as a medium for electron transfer to NAD (P) ⁺ (R. Ruppert, S. et al., Tetrahedron Lett ., 28: 6583, 1987; F. Hollmann, A. et al., Angew. Chem . 113: 190, 2001; Angew. Chem. Int. Ed. , 40: 169 , 2001; K. Vuorilehto et al., Bioelectrochemistry , 65: 1, 20 04) The rhodium complex has been used for the regeneration of FADH ₂ (F. Hollmann et al. Journal of Molecular Catalysis B: Enzymatic 19-20: 167, 2003). However, the electrochemical regeneration method has a disadvantage in that the productivity is limited because the yield is essentially dependent on the surface area of the electrode. The use of a large surface area electrode can achieve such a high reaction rate and productivity, but there is a difficulty in mass production due to an increase in the infrastructure cost.

On the other hand, photochemical regeneration of cofactors has been in the spotlight in recent years in that the solar energy can be used as an unlimited energy source. However, despite these advantages, the regeneration efficiency of the cofactor was not high, because the high efficiency photosensitive agent was not developed. Due to the nature of the photochemical regeneration method that obtains energy from the sunlight through the photosensitive agent, the photosensitive agent should absorb a large amount of energy from the sunlight if possible. TiO ₂ , ZrO ₂ , SrTiO ₃ , Ta ₂ O ₅ and the like were mainly suitable for absorbing light in the ultraviolet region. Given the fact that the energy in the ultraviolet region corresponds to 4% of the total solar energy, the development of a new photoresist that absorbs the visible region, which accounts for 46%, is essential.

On the other hand, nanoparticles have the advantage of being easy to control the optical properties by controlling the particle size, synthesized at a relatively low value at low temperatures, and has a large surface area compared to the volume. Due to these advantages, nanoparticles are used in various fields such as multicolor optical coding, biological imaging, and optoelectronic devices, and among them, CdS, CdSe, and CdTe nanoparticles have been widely used in solar cells that are in the spotlight recently. The high absorption coefficients of these nanoparticles help the solar cells absorb large amounts of light, and their excellent light stability enables long-term stable use of the solar cells.

Thus, the present inventors have confirmed that when the cadmium nanoparticles are used to photochemically regenerate oxidoreductase cofactors, the present invention can efficiently regenerate oxidoreductase cofactors in the visible region, thereby completing the present invention. It became.

It is an object of the present invention to provide a method capable of efficiently regenerating oxidoreductase cofactors in the visible light region.

In order to achieve the above object, the present invention provides an oxidoreductase containing (i) an oxidized oxidoreductase cofactor, (ii) an inorganic photosensitizer, (iii) a redox mediator and (iv) a sacrificial electron donor. Provided is an optical regeneration method of an oxidoreductase cofactor, characterized in that light is irradiated to the cofactor regeneration solution.

The present invention also provides a composition for regeneration of an oxidoreductase cofactor containing (i) an oxidized oxidoreductase cofactor, (ii) an inorganic photosensitizer, (iii) a redox mediator, and (iv) a sacrificial electron donor. To provide.

According to the present invention, it is possible to rapidly regenerate the oxidoreductase cofactor by only irradiating light in the visible light region, and can be very useful for increasing the efficiency of various biocatalytic reactions using the oxidoreductase enzyme.

1 shows a schematic diagram of optical regeneration of NADH using cadmium nanoparticles and rhodium complexes.
2 shows fluorescence spectra and cyclic voltammograms for analyzing the optical regeneration mechanism of NADH.
Figure 3 shows the results of photochemical NADH regeneration experiments with the concentration of the cadmium nanoparticles and whether or not light irradiation.
FIG. 4 compares the cadmium nanoparticle efficiencies in photochemical NADH regeneration systems with the regeneration of NADH using different types of inorganic photosensitizers.
FIG. 5 shows the effect of particle size change of cadmium nanoparticles on the photochemical regeneration of NADH.
6 compares NADH regeneration results of nanoparticles and microparticles under the same surface area.

In one aspect, the present invention is directed to regeneration of an oxidoreductase cofactor containing (i) an oxidized oxidoreductase cofactor, (ii) an inorganic photosensitizer, (iii) a redox mediator, and (iv) a sacrificial electron donor The present invention relates to an optical regeneration method of an oxidoreductase cofactor, which is irradiated with light.

In the present invention, the oxidized oxidoreductase cofactor may be selected from the group consisting of NAD ⁺ , NADP ⁺ , FAD ⁺ and FMN ⁺ , wherein the inorganic photosensitizer is Cd, Ti, Zn, It may be characterized in that it is selected from the group of nanoparticles consisting of W and Ta.

Preferably, the Cd nanoparticles may be CdS, CdSe, CdTe, the Ti nanoparticles may be TiO ₂ , SrTiO _{3 and the} like, Zn nanoparticles may be ZnS, In ₂ O ₃ (ZnO) _{3 and the} like, The W nanoparticles may be WO ₃ , W ₂ Fe ₄ Ta ₂ O _17, or the like, and the Ta nanoparticles may be Ta ₂ O ₅ , NaTaO _3, or the like.

In the present invention, the Cd (cadmium) nanoparticles may be characterized in that the nanoparticles of cadmium crystals selected from the group consisting of CdS (cadmium sulfide), CdSe (cadmium selenide) and CdTe (cadmium telluride). .

In the present invention, it is preferable to use cadmium nanoparticles with nanoparticles having a size of 1 to 5 nm.

In the present invention, the redox mediator may be selected from the group consisting of methylviologen, ruthenium II complex and rhodium III complex, wherein the ruthenium II complex is (hexamethylbenzene-2,2 ' -Bipyridinechloro) ruthenium (II), and the rhodium III complex may be characterized as (pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III). .

In the present invention, the sacrificial electron donor is triethanolamine (TEOA), ethylenediaminetetraacetic acid (EDTA), citric acid, formic acid, ascorbic acid, ascorbic acid, oxalic acid ( Oxalic acid), alcohols and water, and the light may be selected from the group consisting of tungsten-halogen lamp light, xenon lamp light, short wavelength laser light and sunlight. Can be.

In the present invention, it may be characterized in that it is carried out in an oxidoreductase reactor comprising an artificial light source and a quartz reactor.

In another aspect, the present invention is directed to regeneration of an oxidoreductase cofactor containing (i) an oxidative oxidoreductase cofactor, (ii) an inorganic photosensitizer, (iii) a redox mediator, and (iv) a sacrificial electron donor It relates to a composition for.

The present invention provides a photochemical regeneration method of an oxidoreductase cofactor comprising the following two steps.

<Step 1>: sacrificial electron donor in an oxidoreductase reactor; Inorganic photosensitizers that absorb light and are active in redox reactions; Redox mediators; Adding an oxidoreductase cofactor of the oxidized type; And

<Step 2>: the step of irradiating light to the reactor of step 1 to generate a reduced oxidoreductase cofactor.

The reactor is preferably provided with a system comprising a reaction solution containing an inorganic photosensitizer, a redox mediator and a sacrificial electron donor and a light source for irradiating light.

The light source used in the system may preferably be natural sunlight and may be an artificial tungsten-halogen lamp, xenon lamp or short wavelength laser device. In the case of using an artificial light source, the output of the light source is preferably 5 W to 2000 W. If it is lower than the output range, the light intensity is weak, so that the catalytic function may not appear. At too high output, the biocatalytic reaction may not occur or an unwanted reaction may occur due to the degeneration of enzymes and other substances by light and heat.

The sacrificial electron donor is a material that provides electrons to fill the electron pores generated by the transfer of the electrons excited from the inorganic photosensitive agent to the redox mediator, and from the viewpoint of those skilled in the art, Triethanolamine (TEOA), EDTA), Citric acid, Formic acid, Ascorbic acid, Oxalic acid or water, etc. may be used, but preferably TEOA or EDTA.

The inorganic photosensitizer is intended for delivery to the redox mediator in a system in which the redox mediator and the sacrificial electron donor are present 1) a medium for delivering electrons of high energy state to the redox mediator. It acts as a double mediator by functioning as a mediator to obtain the electron from the sacrificial electron donor. From the viewpoint of those skilled in the art, the inorganic photosensitizer may generate an electric current by exciting electrons through light, and may be replaced with any nanoparticle having an energy level suitable for a dual mediator, but preferably CdS, CdSe, CdTe nanoparticles can be used.

The redox mediator is used for the regeneration of the primary mediator for electron transfer to the oxidative oxidoreductase cofactor and a form having activity in the enzymatic reaction of the cofactor. Among the redox mediators, methyl viologen was used for indirect electrochemical regeneration with Flavoenzyme (Ferredoxin reductase (FDR) or Lipoamide Dehydrogenase (LipDH)) as electron transfer mediator for NAD (P) H. (Dicosimo et al. J Org Chem, 46: 4622, 1981) and the ruthenium (II) complex (hexamethylbenzene-2,2′-bipyridinechloro) ruthenium (II) for the reduction of ketones to alcohols. Ogo S, et al., Organometallics 21: 2964, 2002; Yaw Kai Yan et al. J Biol Inorg Chem, 11: 483, 2006), a rhodium (III) complex Pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III): [Cp * Rh (bpy) H ₂ O] ²⁺ (hereinafter M _ox ) for electron transfer to NAD (P) ⁺ Media (R. Ruppert et al. Tetrahedron Lett ., 28: 6583, 1987; F. Hollmann et al. , Angew. Chem ., 113: 190, 2001; Angew. Chem. Int. Ed. , 40: 169, 2001; K. Vuorilehto et al. , Bioelectrochemistry , 65: 1, 2004) and It has been used as a medium for electron transfer to FAD ⁺ (F. Hollmann et al. Journal of Molecular Catalysis B : Enzymatic 19-20: 167, 2003). Thus, the redox mediator is used to improve the regeneration kinetics of oxidoreductase cofactors by delivering electrons and protons.

The ruthenium II complex may preferably be (hexamethylbenzene-2,2′-bipyridinechloro) ruthenium (II), and the rhodium III complex may be (pentamethylcyclopentadienyl-2,2′-bipyridine Preference is given to using chloro) rhodium (III).

However, in the present invention, the presence of the redox mediator is effective in regenerating the cofactor in the form having activity in the enzymatic reaction and speeding up the regeneration of the cofactor, even if the redox mediator does not exist, The presence of redox mediators does not limit the scope of the present invention, as direct chemical photoregeneration of cofactors is possible.

The oxidoreductase cofactor of the oxidized type is nicotinamide cofactor NAD ⁺ (nicotinamide adenine dinucleotide), NADP ⁺ (nicotinamide adenine dinucleotide phosphate) or flavin cofactor FAD ⁺ (flavin adenine dinucleotide), FMN ⁺ (flavin mononucleotide) yl And preferably NAD ⁺ .

Optical regeneration method of the oxidoreductase cofactor of the present invention, as shown in Figure 1, the inorganic photosensitive agent absorbs light to excite the electrons and transfer the excited electrons to the redox mediator (step a); The step (a) of the inorganic photosensitive agent of step a being supplemented with electrons from the sacrificial electron donor (step b); The redox mediator receiving the electrons and protons passes the electrons and protons to the oxidative oxidoreductase cofactor to form a reduced oxidoreductase cofactor (step c).

In one aspect of the present invention using a cadmium nanoparticles, inorganic light-sensitive agent, and was used as a rhodium complex as a redox mediator, and in this case a rhodium complex M ₁ is through the electrochemical / chemical process is converted to the active reduction chain M ₃ NADH Is involved in the regeneration of. M ₁ accepts two electrons and becomes an electrochemical change into M ₂ (E-step). The M ₂ is then converted to M ₃ through a chemical process by taking one proton in solution without changing the total amount of electrons. The active reductant M ₃ provides two electrons and one proton to NAD (P) ⁺ to convert to NAD (P) H, where it returns to its initial state M ₁ .

In order to confirm the effect of the present invention, the change over time of the reproducible NADH yield using CdS, CdSe, CdTe nanoparticles was observed. As a result, it was confirmed that NADH is not regenerated when irradiated with light, but is regenerated at the same time as irradiated with light, and the yield of regenerated NADH is increased according to the concentration of nanoparticles used. In addition, compared with the case of regenerating NADH using other types of inorganic photosensitizers, CdS, CdSe, and CdTe nanoparticles showed significantly higher efficiency than other inorganic photosensitizers. In addition, the size control of the nanoparticles and the comparison with the CdS, CdSe, CdTe microparticles observed that the smaller the size of the NADH regeneration yield increases. (See Experimental Example 2, Experimental Example 3)

From this, the cadmium nanoparticles used in the present invention act as a catalyst active by light in the presence of a sacrificial electron donor and participate in the transfer of protons and electrons, thereby contributing to increasing the regeneration rate of the oxidoreductase cofactor (FIG. 3).

The concentration of the cadmium nanoparticles contained in the composition for oxidoreductase cofactor regeneration of the present invention is preferably 1 to 50 mM, more preferably 5 to 20 mM.

Therefore, the method of the present invention can increase the efficiency of various redox catalysis reactions using oxidoreductase enzymes, and is particularly effective under poor environments such as a high pH environment. It is also contemplated that the method of the present invention can be extended not only for the regeneration of oxidoreductase cofactors but also for chemical reactions of other substances.

Hereinafter, the present invention will be described in more detail with reference to examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example: Regeneration of NADH by Photochemical Method Using Cadmium Nanoparticles and Rhodium Composites

In this embodiment, a rhodium complex (pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III) as an organometallic medium for transferring electrons between NAD ⁺ and the electrode (hereinafter referred to as M; [Cp ^* Rh (bpy) Cl] ⁺ ; Cp ^* = C ₅ Me ₅ , bpy = 2,2'-bipyridine).

The rhodium complex M was synthesized by the method of Kolle and Grazel (F. Hollmann et al, J. Mol. Catal . B , 19-20 : 167, 2002). In detail, a solution of rhodium precursor (RhCl ₃ H ₂ O, 200 mg) and hexamethyl dewarbenznen (40 mg) prepared in 6 ml of methanol was refluxed at 65 ° C. under a nitrogen atmosphere for 15 hours. The reaction solution was then cooled to room temperature, the solvent was evaporated in vacuo, washed with ether to remove the excess hexamethyl diwarbenzene and extracted with chloroform to give a red precipitate. Magnesium sulphate was used to remove moisture from the solution, the solvent was evaporated at low pressure, and the remaining was recrystallized from chloroform-benzene. Mixing with 2 equivalents of 2.2'-bipyridine in a methanol solvent gives a yellow solution, from which [Cp * Rh (bpy) Cl] Cl is used with diethyl ether. Was recrystallized to finally obtain a rhodium complex.

In order to regenerate NADH by the photochemical method, an artificial light source and a quartz reactor capable of irradiating light therefrom were fabricated. As an artificial light source, a tungsten-halogen lamp having a power of 400 W was used, and in particular, a filter for filtering light having a wavelength of 420 nm or less to irradiate light in the visible region was mounted. In addition, in order to exclude the result that can be obtained from the heat of the light source, a filter capable of filtering light having wavelengths in the heat and infrared regions is constructed using cooling water. Light from the light source was irradiated to the reaction solution contained in the quartz reactor.

Rhodium complex M, a redox mediator for NADH production, was used at a concentration of 0.25 mM, and cadmium nanoparticles used as a catalyst that absorbed light and was active in redox reactions were used at concentrations of 2 mM, 10 mM or 20 mM. NAD ⁺ was used at a concentration of 1 mM. Phosphate buffer (100 mM, pH 7.0) was used as an electrolyte, and the sacrificial electron donor TEOA, cadmium nanoparticles, M and NAD ⁺ were added to the buffer, dissolved by stirring, and irradiated with light to regenerate NADH.

Experimental Example 1 Measurement of Fluorescence Spectrum and Cyclic Voltammetry for Analysis of Optical Regeneration Mechanism of NADH

To analyze the optical regeneration mechanism of NADH, fluorescence spectra were analyzed and cyclic voltammetry was measured.

First, 2mM CdTe nanoparticles were dissolved in phosphate buffer (100mM, pH 7.0), and then the fluorescence spectra of various concentrations of the rhodium complex M were measured (FIG. 2A). The fluorescence intensity of the CdTe nanoparticles decreased in proportion to the concentration of the rhodium complex M which was put together, which transferred the electrons to M instead of fluorescing when the excited electrons fall back to the ground state. It is because of this phenomenon. Therefore, as the concentration of M increases, the number of excited electrons transferred to M increases, thereby decreasing the fluorescence intensity. The electrons transferred to M are transferred to NAD ⁺ .

After dissolving 2 mM CdTe nanoparticles and 0.25 mM rhodium complex M in phosphate buffer (100 mM, pH 7.0), the circulating voltage of CdTe nanoparticles and M according to the presence or absence of NAD ⁺ (concentration of 0 mM or 1 mM) The amperage was compared.

As a result, as shown in (b) of Figure 2, the reduction peak (reduction peak current) of the CdTe nanoparticles and M was significantly increased when the presence of NAD ⁺ than when not present. This evidence sikyeotdaneun promote CdTe nanoparticles and M a reduction of the NAD ^+, the electron transfer from the end CdTe nanoparticles M is passed to NAD ⁺ means a reduced-rescue the NAD ⁺ to NADH.

In the case of CdS and CdSe nanoparticles, experiments under the same conditions as CdTe nanoparticles showed the same tendency as CdTe nanoparticles as shown in (c)-(f) of FIG. 2. Therefore, it can be seen from the experimental results of FIG. 2 that the optical reproduction schematic of the NADH of FIG. 1 is correct.

Experimental Example 2: Measurement of NADH Regeneration Rate According to Concentration of Cadmium Nanoparticles and Irradiation of Light

The change over time of the NADH yield reproduced using CdS, CdSe, and CdTe nanoparticles was specified (FIG. 3). Detailed experimental conditions are as follows. Cadmium nanoparticles were used at a concentration of 2mM, 10mM or 20mM, rhodium complex M was used at a concentration of 0.25mM, NAD ⁺ was used at a concentration of 1mM. Phosphate buffer (100 mM, pH 7.0) was used as an electrolyte, and the sacrificial electron donor TEOA, cadmium nanoparticles, M and NAD ⁺ were added to the buffer, dissolved by stirring, and irradiated with light to regenerate NADH.

As a result, as shown in Figure 3, NADH is not regenerated if the light is not irradiated and is reproduced at the same time irradiated with the light, the NADH yield increased according to the concentration of the nanoparticles used. NADH yields measured after 60 minutes were 52% (CdS nanoparticles, 20 mM), 29% (CdSe nanoparticles, 20 mM) and 55% (CdTe nanoparticles, 20 mM), respectively, which are different from FIG. 4. Compared to the regeneration of NADH using inorganic photosensitizers. CdS, CdSe and CdTe nanoparticles showed significantly higher efficiency than other photosensitizers. Among them, the turnover frequency of CdTe nanoparticles was 180 times higher than that of P-doped TiO ₂ and W ₂ Fe ₄ Ta ₂ O ₁₇ . 270 times higher.

Experimental Example 3: Effect of Particle Size Change of Cadmium Nanoparticles on Photochemical Regeneration of NADH

In general, the properties of nanoparticles are determined by particle size, particle composition, particle shape, etc., which affects the photoefficiency and the number of surface active sites of the nanoparticles. Therefore, the particle size change may affect the photochemical regeneration of the NADH, and to confirm this, the NADH was regenerated using CdSe nanoparticles and microparticles of various sizes.

CdSe nanoparticles with different particle sizes and rhodium complexes M and NAD ⁺ were dissolved in phosphate buffer (100 mM, pH 7.0) at concentrations of 10 mM, 0.25 mM and 1 mM, respectively, and then sacrificial electron donor TEOA was added to the buffer and Was investigated to regenerate NADH.

As a result, as shown in Figure 5, the smaller the particle size was observed that the regenerated NADH yield is higher. This means that the smaller the nanoparticles, the faster the movement of electrons and holes, so that the charge transfer rate from the nanoparticles to the rhodium complex M is the electron-hole recombination rate. Because it is faster. In addition, the smaller nanoparticles are more active because the number of surface active sites, especially corner sites and edge sites, available in the actual reaction increases. To prove this, NADH was regenerated using cadmium nanoparticles and cadmium microparticles under the same surface area (0.1 m ² ).

As a result, as shown in FIG. 6, the microparticles did not regenerate NADH, whereas the nanoparticles regenerated NADH. This is because nanoparticles have a much larger number of corner sites and edge sites than microparticles.

The specific parts of the present invention have been described in detail above, and it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (18)

(i) an oxidized oxidoreductase cofactor selected from the group consisting of NAD ⁺ , NADP ⁺ , FAD ⁺ and FMN ⁺ , (ii) CdS (cadmium sulfide), CdSe (cadmium selenide) and CdTe (cadmium telluride) Inorganic photosensitive agent consisting of nanoparticles of cadmium crystal selected from the group consisting of (iii), (iii) redox mediator and (iv) oxidoreductase cofactor regeneration solution containing a sacrificial electron donor. Optical regeneration method of an oxidoreductase cofactor.
delete delete delete The method of claim 1, wherein the redox mediator is selected from the group consisting of methylviologen, ruthenium II complex, and rhodium III complex.
The optical regeneration method of an oxidoreductase cofactor according to claim 5, wherein the ruthenium II complex is (hexamethylbenzene-2,2'-bipyridinechloro) ruthenium (II).
6. The method of claim 5, wherein the rhodium III complex is (pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III).
The method of claim 1, wherein the sacrificial electron donor is triethanolamine (TEOA), ethylenediaminetetraacetic acid (EDTA), citric acid, formic acid, ascorbic acid, oxalic acid (Oxalic acid), an alcohol and an optical regeneration method of oxidoreductase cofactors selected from the group consisting of water.
The optical regeneration method of an oxidoreductase cofactor according to claim 1, wherein the light is selected from the group consisting of tungsten-halogen lamp light, xenon lamp light, short wavelength laser light and sunlight.
The optical regeneration method of an oxidoreductase cofactor according to claim 1, which is performed in an oxidoreductase reactor including an artificial light source and a quartz reactor.
(i) an oxidized oxidoreductase cofactor selected from the group consisting of NAD ⁺ , NADP ⁺ , FAD ⁺ and FMN ⁺ , (ii) CdS (cadmium sulfide), CdSe (cadmium selenide) and CdTe (cadmium telluride) An inorganic photosensitizer composed of nanoparticles of cadmium crystals selected from the group consisting of: (iii) a redox mediator and (iv) a oxidoreductase cofactor regenerating composition containing a sacrificial electron donor.
delete delete delete 12. The composition for oxidoreductase cofactor regeneration according to claim 11, wherein the redox mediator is selected from the group consisting of methylviologen, ruthenium II complex and rhodium III complex.
16. The composition of claim 15, wherein the ruthenium II complex is (hexamethylbenzene-2,2'-bipyridinechloro) ruthenium (II).
The oxidoreductase cofactor regenerating composition according to claim 15, wherein the rhodium III complex is (pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III).
The method of claim 11, wherein the sacrificial electron donor is triethanolamine (TEOA), ethylenediaminetetraacetic acid (EDTA), citric acid, formic acid, ascorbic acid, oxalic acid (Oxalic acid), oxidoreductase cofactor regeneration composition, characterized in that selected from the group consisting of alcohols and water.

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