KR20130118001A - Method for photochemical reduction of nad(p) analogs and method of biocatalyzed artificial photosynthesis using the same - Google Patents

Abstract

PURPOSE: An artificial photosynthesis method by photochemical regeneration of an NAD(P) derivative is provided to produce eco-friendly fine chemicals and a raw material for a novel drug. CONSTITUTION: A photochemical regeneration method of an NAD(P) derivative comprises the steps of: irradiating a regeneration solution by light; and generating a useful material produced by oxidoredutase reaction using the regenerated NAD(P) derivatives. The regeneration solution contains an NAD(P) derivative, a dye, a sacrificial electron donor, and an electron mediator.

Description

Photochemical Reduction of NAD Derivatives and Method of Artificial Photosynthesis Using The Same Method for Photochemical Reduction of NAD (P) Analogs and Method of Biocatalyzed Artificial Photosynthesis Using the Same

The present invention relates to photochemical regeneration of NAD (P) derivatives and artificial photosynthesis method using the same, and more particularly, to regenerate coenzymes using NAD (P) derivatives, and to use the regenerated coenzymes for redox enzyme reactions. It relates to an artificial photosynthesis method for preparing a useful material.

Nicotinamide coenzymes are involved in the electron transfer of the redox reaction catalyzed by oxidoreductase in vivo. The reduced nicotinamide coenzyme (ie, NAD (P) H) is useful because it can be used in various enzyme reactions such as hydroxylation, epoxidation, Bayer-Villiger, etc. . On the other hand, NADH plays a very important role in performing enzymatic reactions because it acts as a redox equivalent in biocatalytic reactions using redox enzymes. However, coenzymes, 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 coenzyme is quite expensive, there are many economic difficulties in using it in the actual industry. Therefore, it is very important to efficiently regenerate coenzymes from NAD (P) ⁺ consumed during the redox reaction.

In order to increase the efficiency of the biocatalyst reaction, coenzymes for the continuous reaction of the enzymes need to be continuously regenerated, but unlike the hydrolytic enzymes that are widely used in various ways, they are satisfactory in the use of redox enzymes. The coenzyme regeneration method has not been established, so it is not widely commercialized. Electrochemical regeneration has thus been considered as an attractive alternative to the existing second enzyme / substrate regeneration (F. Hollmann and A. Schmid, Biocatal . Biotransform . , 22:63, 2004). . However, in the electrochemical regeneration method, the reduction of NAD (P) ⁺ to NAD (P) H was also disadvantageous in that the regeneration efficiency was lowered due to the slow electron transfer rate between the electrode and NAD (P) ⁺ even under the thermodynamically favorable voltage condition.

To solve this problem, a method of transferring electrons between an electrode and NAD (P) ⁺ using a homogeneous electron mediator has been developed (H. Jaegfeldt et al., Anal . Chem. , 53: 1979, 1981; J. Wang and J, Liu Anal , Chim . Acta , 284: 385, 1993; J. Wang et al., Anal . Chim . Acta , 360: 171, 1998). 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 allows for higher reaction rates and higher productivity, but increases the cost of infrastructure.

On the other hand, the photochemical approach to coenzyme regeneration has attracted attention recently because it can utilize vast solar energy in a manner similar to the photosynthesis of nature. Simulating nature's photosynthesis enables low-cost and environmentally friendly photochemical coenzyme regeneration using infinite solar energy without supplying external power (D.Gust, TA Moore, AL Moore, Acc . Chem . Res . 2009, 42, 1890-1898., D. Gust, TA Moore, AL Moore, Acc . Chem . Res . 2001, 34, 40-48). Natural photosynthesis absorbs light from a photosystem consisting of organic dyes such as chlorophyll to obtain high energy electrons, reducing ATP (Adenosine Triphosphate) cofactors and NAD (Nicotinamide Adenosine Dinucleotide) cofactors After regeneration, it is used as a raw material for glucose synthesis. In contrast, in vitro, man-made photosensitizers are used instead of chlorophyll to absorb sunlight and generate high-energy photo-excited electrons that can be used for redox enzyme reactions. Cofactors are photochemically regenerated. However, these photochemical reduction methods still exhibit low conversion efficiencies, and the reaction mechanisms are very difficult and therefore difficult to implement.

Accordingly, the present inventors have made diligent efforts to solve the above problems, and as a result of using the NAD (P) derivative having a more positive redox potential instead of the existing NAD (P), the coenzyme can be regenerated with excellent efficiency. In addition, it was confirmed that useful substances can be generated by using the regenerated coenzyme in the redox enzyme reaction, thereby completing the present invention.

It is an object of the present invention to provide a high efficiency photochemical NAD (P) derivative regeneration method.

Another object of the present invention is to provide a composition for regenerating a NAD (P) derivative.

It is still another object of the present invention to provide an artificial photosynthesis method using photochemical regeneration of a NAD (P) derivative.

In order to achieve the above object, the present invention (i) NAD (P) derivative; (ii) dyes; (iii) sacrificial electron donors; And (iv) irradiating light to a regeneration solution containing an electron mediator.

The present invention also provides for (i) a NAD (P) derivative; (ii) dyes; (iii) sacrificial electron donors; And (iv) a composition for regeneration of a NAD (P) derivative containing an electron mediator.

The present invention also relates to a regeneration solution containing (a) (i) a NAD (P) derivative, (ii) a dye, (iii) a sacrificial electron donor and (iv) an electron mediator to irradiate the NAD (P) derivative with light. Photochemical regeneration; And (b) using the regenerated NAD (P) derivative in a redox enzymatic reaction to produce a useful material produced by the redox enzymatic reaction. It provides a photosynthesis method.

According to the present invention, if the NAD (P) derivative is reduced by photochemical method, it is useful to increase the efficiency of various biocatalytic reactions using the same, since it can reduce a larger amount than NAD (P) at a high speed. In addition, according to the artificial photosynthesis method using photochemical regeneration of the NAD (P) derivative, it is useful for environmentally friendly production of high value-added fine chemicals such as new drug materials and optical isomers.

Figure 1 shows a schematic diagram that the electrons photo-electron by the photosensitive agent through the absorption of visible light causes the reduction of the rhodium complex (M) together with the photochemical reduction of the NAD and NAD derivatives.
Figure 2 shows the result of the regeneration of the coenzyme using the NAD (P) derivative, (a) shows the visible light-induced reduction concentration of NAD and its derivatives, (b) shows the yield of enzymatically active coenzyme It is shown.
Figure 3 shows the relative positions of the cathodic peak potential of NAD and its derivatives.
Figure 4 (a) shows the effect of NAD and its derivatives on the M-dependent quenching of EY fluorescence, (b) shows the change in fluorescence spectrum of EY in the presence of NAD or TNAD.
Figure 5 relates to the photoenzymatic synthesis of L- glutamate, (a) shows the effect of the coenzyme concentration on the conversion rate, (b) shows the coenzyme concentration on the turnover frequency (TOF) of the coenzyme The effect is shown.
Figure 6 shows the turnover number (TON) of Eosin Y in photochemical regeneration of the coenzyme.
Figure 7 shows the change in absorption spectrum upon photochemical reduction of NAD and its derivatives in the presence / absence of EY.
8 (a) shows a cyclic voltammetry diagram of M and a coenzyme derivative, and (b) shows a cyclic voltammetry diagram of M in the presence / absence of coenzyme.
Figure 9 shows the conversion rate of the photoenzymatic reaction of GDH according to the concentration of the coenzyme and EY.
Figure 10 shows the absorption spectrum of NADH to confirm the stability of the NADH.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

The present invention (i) NAD (P) derivatives; (ii) dyes; (iii) sacrificial electron donors; And (iv) relates to a photochemical regeneration method of the NAD (P) derivative and the composition for regeneration of the NAD (P) derivative, characterized in that the light is irradiated to the regeneration solution containing the electron mediator.

As used herein, "NAD (P)" means "NAD or NADP" and "NAD (P) H" means "NADH or NADPH".

NAD and NADH both exhibit the same coenzyme activity that transfers electrons and protons, except that NAD or NADP is used as coenzyme, depending on the function and type of the redox enzyme. Although the two coenzymes show structural differences, NAD and NADP have common photochemical reduction properties because the photochemical reduction reaction in the present invention is a nicotinamide ring portion bonded to ribose. Can be seen. According to an embodiment of the invention, four commercially available NAD derivatives having three substituted pyridine rings, 3-acetylpyridine adenine dinucleotide, APAD, R = COCH ₃ ), 3-pyridinealdehyde adenine dinucleotide (PAAD, R = CHO), thio-nicotinamide adenine dinucleotide (TNAD, R = CSNH ₂ ) and nicotinic acid adenine dinucleotide ( nicotinic acid adenine dinucleotide, NAAD, R = COOH). The compounds may be synthesized by general chemical synthesis methods known in the art, and commercially available compounds may be purchased and used.

As can be seen through the following examples, NAD (P) derivatives according to the present invention can reduce more than NAD (P) when the photochemical reaction using the same dye, NAD (P) H It is characterized in that excellent synthesis efficiency can be obtained in the photoenzymatic synthesis reaction of the dependent enzyme reaction product.

In the present invention, the dye may be any dye capable of generating an electric current by excitation of electrons through light, but preferably includes fluorescein. Substituted Eosin Y, Eosin B, Erythrosine B, Phloxine B, Rose Bengal, Merbromin, Ethyl One or more selected from the group consisting of Ethyl Eosin, Rhodamine B, Rhodamine 6G, and Sulforhodamine B may be used, and more preferably, Eosin Y Can be used. Eosin Y was used as a photosensitizer because it could reduce NAD by visible light irradiation.

Although the addition amount of the said dye is not restrict | limited significantly, Preferably it is added at the density | concentration of 0.1 micrometer-100 micrometers, More preferably, it is good to add it at the density | concentration of 1 micrometer-100 micrometers.

In the present invention, the sacrificial electron donor is a material that provides electrons to fill the electron pores generated by the transfer of electrons excited from the dye to an electron mediator. In view of those skilled in the art, triethanolamine (Triethanolamine, TEOA), ethylenediaminetetraacetic acid (EDTA), citric acid, formic acid, ascorbic acid, ascorbic acid, oxalic acid, alcohols or water, etc. May be TEOA.

The electron mediator used in the present invention may be any one or more selected from the group consisting of methyl viologen, ruthenium (II) complex, and rhodium (III) complex, and the electron mediator is an oxidized type. It is used as a primary mediator for electron transfer to oxidoreductase cofactors. In general, direct electron transfer in which electrons are transferred directly to an oxidative oxidoreductase cofactor without passing through an electron mediator results in the formation of isomers and dimers of the cofactor. It is not enzymatically inactive to obtain the final product in redox enzyme reactions.

The ruthenium (II) complex may preferably be (hexamethylbenzene-2,2'-bipyridinechloro) ruthenium (II), and the rhodium (III) complex may be (pentamethylcyclopentadienyl-2,2 Preference is given to using '-bipyridinechloro) rhodium (III). Rhodium (III) complexes are widely used for indirect regeneration of NAD (P) H, which is active in enzymatic reactions because they maintain high regioselectivity (> 99%) even with pH and temperature changes.

In the photochemical reduction reaction, electrons are released from the sacrificial electron donor by the visible light absorption of the photosensitizer. The released electrons are transferred to the oxidized mediator (M _ox ) and accept protons in aqueous solution to become the reduced form (M _red ). Hydrogen (ie two electrons and one proton) is transferred from M _red to the oxidized nicotinamide coenzyme through the catalysis of the medium (FIG. 1).

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 present invention, a reduced oxidoreductase coenzyme is produced by irradiating light with a light source and stirring the NAD (P) derivative regeneration solution containing an oxidized oxidoreductase coenzyme, a dye, a sacrificial electron donor, and an electron mediator. It is preferable that the output of the said light source is 5W-2000W. 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.

In order to confirm the effect of the present invention, another aspect of photochemical reduction of NAD and NAD derivatives was examined using linear sweep voltammetry (LSV). The reduction peak potential ( E _p ) of NAD and NAD derivatives was determined by measuring their reducibility using LSV with the glassy carbon electrode as the working electrode. As shown in FIG. 3, the cathodic peak potential of NAD was determined to be -1.10 V compared to Ag / AgCl, but other derivatives were more positive E _p (APAD = -1.00 V, PAAD = -0.93 V and TNAD = -1.05 V) or More negative E _p (NAAD = -1.28 V). Since NAD derivatives having a high E _p are more easily reduced than NAD, APAD and PAAD have high reducing power.

In another aspect, the present invention is directed to a regeneration solution containing (a) a (i) NAD (P) derivative, (ii) a dye, (iii) a sacrificial electron donor, and (iv) an electron mediator to irradiate a NAD (P) Photochemically regenerating the derivative; And (b) using the regenerated NAD (P) derivative in a redox enzymatic reaction to produce a useful material produced by the redox enzymatic reaction. It relates to a photosynthesis method.

Artificial photosynthesis of the present invention refers to a process of reproducing coenzymes by mimicking the light reaction of natural photosynthesis, and preparing useful substances by enzymatic reaction using redox enzymes by mimicking the cancer reaction. In the present invention, useful materials produced by artificial photosynthesis include new drug materials, such as glutamate, aspartate, ibuprofen, pravastatin, Taxol, Taxol, and the like. High value-added fine chemicals can be exemplified, but includes all substances produced by a redox enzyme reaction requiring NAD (P) or a derivative thereof as a coenzyme.

Artificial photosynthesis of the present invention is characterized by using a NAD (P) derivative as an electron transporter for photoenzymatic synthesis under visible light, and the dye may be any dye that can generate an electric current by exciting electrons through light. However, Eosin Y, Eosin B, and Erythrosine B, which preferably contain fluorescein and have some hydrogen elements substituted with halogen elements as a basic structure. ), Phloxine B, Rose Bengal, Merbromin, Ethyl Eosin, Rhodamine B, Rhodamine 6G and Sulfo At least one selected from the group consisting of multi-amine B (Sulforhodamine B) may be used, and more preferably, Eosin Y may be used.

The sacrificial electron donor is triethanolamine (TEOA), ethylenediaminetetraacetic acid (Ethylenediaminetetraacetic acid, EDTA), citric acid, formic acid, ascorbic acid, ascorbic acid, oxalic acid, alcohol It is characterized in that it is selected from the group consisting of water or water.

The electron mediator is any one or more selected from the group consisting of methylbiologen, ruthenium (II) complex and rhodium (III) complex, and the rhodium (III) complex is (pentamethylcyclopentadienyl -2,2'bipyridinechloro) rhodium (III): [Cp * Rh (bpy) H ₂ 0] ⁺ .

The redox enzyme may be any enzyme that catalyzes a redox reaction using a coenzyme among enzymes of an oxidoreductase group (Oxidoreductase, EC 1), and GDH (glutamate dehydrogenase), ADH (alcohol dehydrogenase), and G6PDH ( glucose-6-phosphate dehydrogenase (LDH), lactic dehydrogenase (LDH), malate dehydrogenase (MDH), and succinic dehydrogenase (SDH), and the like, but are not limited thereto.

By calculating the ratio of enzymatically active coenzyme to total coenzyme amount to determine whether the photochemical reduction products (ie, NADH, APADH, PAADH, TNADH and NAADH) have enzymatic activity for electron transfer to oxidoreductase The yield of active coenzyme was calculated. As expected, the photochemically reduced NAD (ie, NADH) reacted almost 100% with GDH. APADH and PAADH showed approximately 80% active coenzyme yield slightly lower than NADH. For TNAD it was lower than 10% and for NAAD the degree of reduction was too low to measure the yield of active coenzymes. Quantum yields of NAD, APAD and PAAD were calculated to be 0.035, 0.053, and 0.030, respectively. The quantum yield (Φ _coenzyme ) of NAD and its derivatives is calculated as follows.

Φ _coenzyme = 2 * (number of reduced coenzymes / number of incident photons)

[Example]

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  One: NAD (P) Properties of Derivatives

In the present invention, as the control group NAD and the experimental group NAD derivative 3-acetylpyridine adenine dinucleotide (APAD, R = COCH ₃ ), 3-pyridinealdehyde adenine dinucleotide (PApy, R = CHO), thio-nicotinamide adenine dinucleotide (TNAD, R = CSNH ₂ ) and nicotinic acid adenine dinucleotide (NAAD, R = COOH) were used. All compounds including Eosin Y, TEOA, NAD, APAD, PAAD, TNAD and NAAD were purchased from Sigma Aldrich (USA) and used without further purification. Rhodium complex M was synthesized by a known method.

As shown in Figure 2, each coenzyme has a different reducing mode. Compared with NAD, APAD and PAAD showed higher reduction yields. APAD and PAAD showed high yields of 77% and 56%, respectively, after 30 minutes of photo-sensitization by EY, but only 48% of NAD showed reduced yields. In addition, APAD was found to be faster than NAD. In contrast, 8.6% for TNAD and 2.1% for NAAD.

Example  2: NAD (P) photochemical reduction of derivatives

Photochemical reduction of NAD and NAD derivatives was carried out in a quartz reactor equipped with a temperature controlled bath (37 ° C.). A 450 W xenon lamp equipped with a 420 nm cut-off filter was used as the light source. The EY-sensitized photoreduction of nicotinamide coenzyme is directed to a quartz reactor containing a reaction mixture consisting of 3 μM EY, 0.25 mM M, 1 mM coenzyme, 15% w / v TEOA and phosphate buffer (100 mM, pH 7.5). This was done by examining. Using the known extinction coefficient, the concentration of the reduced cofactor at the maximum absorption wavelength (λ max) was obtained (FIG. 7).

NAD and its derivatives were photochemically reduced via M, the electron mediator. Therefore, the interaction of M and coenzyme is very important in the efficiency of photochemical regeneration. In order to determine the effect of M on the electrochemical reduction of NAD, the catalytic activity of M on NAD derivatives was measured by cyclic voltammetry. The cyclic voltammogram of M depending on the presence of NAD and its derivatives was recorded (FIG. 8). Table 1 shows the cathodic peak potential ( E _p ) and cathodic peak current ( I _p ) of M. According to the above results, I _p of M is affected by the presence of the coenzyme, it can be seen that the reduction is successfully mediated by M. APAD increased the I _p of M over NAD. In the presence of TNAD, the reduction peak of M _ox disappeared and instead another cathodic peak was observed at -1.05 V relative to Ag / AgCl. Coordination of M and NAD is known to occur through the oxygen atoms of the Cp * Rh metal center and the amide carbonyl group (CONH ₂ ) of NAD. TNAD has a sulfur atom instead of an oxygen atom of the amide group of NAD, which is more polar and coordinating than oxygen. TNAD's high affinity for Mox leads to the formation of noncatalytic complexes and inhibits photoinduced electron transfer between EY and Mox. NAAD does not increase I _p , which means that M has little catalytic effect on NAAD. Although NAAD has an oxygen atom that can interact with M, unlike TNAD, the NAAD COOH group is dehydrogenated under neutral conditions because the pKa of NAAD is known as 4.77 (37 ° C). NAAD thus binds to the metal center of M via a carboxyl (COO-) group as an ionic bond, which is pyridinyl C ₄ Prevents coordination of carbonyl-O-Rh for hydrogen transfer to atoms.

Cathodic peak potential ( E _p ) and current ( I _p ) of M in the presence / absence of NAD and NAD derivatives E _p / V I _p / μA Catalytic effect ^b M -0.727 -16.3 NA M + NAD -0.768 -17.8 O M + APAD -0.757 -20.4 O M + PAAD -0.757 -17.5 O M + TNAD n.d -5.96 ^a X M + NAAD -0.808 -14.8 X

^a I _p of M + TNAD is determined at -0.757 V

^b NA (not available), O (with catalytic effect), X (without catalytic effect)

The interaction of the photosensitizer with M is as important as the M-coenzyme interaction in photoinduced reduction of the coenzyme according to FIG. 1. In the last study, it was found that the fluorescence intensity of EY decreases with increasing concentration of M due to the stern-volmer relationship (SH Lee et al., Chem Bio Chem , 10: 1621, 2009). The result is due to the reversible binding of EY and M to form the EY-M complexes with the ionized carboxyl groups of the metal centers and EY of M (Rh ^{2 +).} The inventors looked at the effect of NAD derivatives on EY-M while monitoring the change in fluorescence intensity of EY with increasing concentration of M. As shown in FIG. 4 (a), unlike other coenzymes, TNAD prevented the migration of photoexcitation electrons of EY to M, thereby significantly reducing quenching by M. 4 (b) shows that this inhibition is due to the direct interaction of TNAD with EY. The fluorescence intensity of EY decreases with the addition of TNAD, but does not affect other coenzymes. This shows that the direct interaction between TNAD and EY is related to the low conversion of the photoinduced reduction reaction of TNAD shown in FIG. 2.

Example  3: NAD (P) redox enzyme reaction of derivative

Photochemically regenerated coenzymes can be used for redox enzyme reactions, and in this embodiment L-glutamate using L-glutamate dehydrogenase (GDH) as an oxidoreductase. ). Photoenzymatic synthesis of L-glutamate was performed in a phosphate buffer (100 mM, pH 7.5) containing 15% w / v of TEOA, 5 μM of EY, 0.25 mM M, 0.1-0.5 mM photoreduced coenzyme, 10 mM α -Ketoglutarate, 200mM ammonium sulfate and 100 U GDH in the reaction solution.

In Figure 4 (a), the initial reaction conversion at 20 minutes is shown for the concentration of oxidized coenzyme used for the reaction. Reaction conversion reached a plateau of conversion at concentrations above 0.3 mM of NAD, which appears to be due to other limiting factors (eg, EY) rather than NAD. In fact, the conversion of reaction at 0.5 mM NAD is enhanced when the EY increases to 20 μM (FIG. 9). According to this result, the reaction is completely converted at reaction times longer than 20 minutes depending on the EY concentration and the type of coenzyme. The inventors found that using APAD instead of NAD significantly improved the photoenzymatic L-glutamate synthesis reaction (FIG. 5). PAAD (FIG. 2 and Table 1), which is less effective at reducing than NAD, shows low enzymatic conversion rates. According to the turnover frequency (TOF) of each coenzyme of FIG. 5, APAD shows higher TOF in all coenzyme concentration ranges than NAD. For example, the TOF of APAD is 1.3 times higher than NAD (25h ⁻¹ ) at 33 h ⁻¹ at 0.1 mM (FIG. 4 (b)).

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (20)

(i) NAD (P) derivatives; (ii) dyes; (iii) sacrificial electron donors; And (iv) irradiating light to a regeneration solution containing an electron mediator. The method of claim 1, wherein the NAD (P) derivative is APAD or PAAD. The method of claim 1, wherein the dye is Fluorescein (Fluorescein), Eosin Y (Eosin Y), Eosin B (Eosin B), Erythrosine B (Phloxine B), Rose Bengal (Rose Bengal), Merbromin, Ethyl Eosin, Rhodamine B, Rhodamine 6G, and Sulforhodamine B Photochemical regeneration method of the NAD (P) derivative, characterized in that. 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 a photochemical regeneration method of the NAD (P) derivative, characterized in that selected from the group consisting of water. The photochemical regeneration method of 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 photochemical regeneration method of NAD (P) derivative according to claim 1, wherein the electron mediator is selected from the group consisting of methylbiologen, ruthenium II complex, and rhodium III complex. 8. The method of claim 6, wherein the rhodium III complex is (pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III). (i) NAD (P) derivatives; (ii) dyes; (iii) sacrificial electron donors; And (iv) a composition for regeneration of a NAD (P) derivative containing an electron mediator. 9. The composition for regenerating a NAD (P) derivative according to claim 8, wherein the NAD (P) derivative is APAD or PAAD. The method of claim 8, wherein the dye is Fluorescein (Fluorescein), Eosin Y (Eosin Y), Eosin B (Eosin B), Erythrosine B (Phloxine B) , Group consisting of Rose Bengal, Merbromin, Ethyl Eosin, Rhodamine B, Rhodamine 6G and Sulforhodamine B The composition for regeneration of NAD (P) derivative, characterized in that selected from. The method of claim 8, wherein the sacrificial electron donor is triethanolamine (TEOA), ethylenediaminetetraacetic acid (EDTA), citric acid, formic acid, ascorbic acid, oxalic acid (Oxalic acid), a composition for regeneration of a NAD (P) derivative, characterized in that selected from the group consisting of alcohol and water. 9. The composition for regenerating a NAD (P) derivative according to claim 8, wherein the electron mediator is selected from the group consisting of methylviologen, ruthenium II complex, and rhodium III complex. The composition for regeneration of a NAD (P) derivative according to claim 12, wherein the rhodium III complex is (pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III). Artificial photosynthesis method using photochemical regeneration of NAD (P) derivative comprising the following steps:
(a) photochemical regeneration of the NAD (P) derivative by irradiation of light to (i) a NAD (P) derivative, (ii) a dye, (iii) a sacrificial electron donor, and (iv) an electron mediator. step; And
(b) using the regenerated NAD (P) derivative in a redox enzyme reaction to prepare a useful material produced by the redox enzyme reaction. 15. The method of claim 14 wherein the NAD (P) derivative is APAD or PAAD. The method of claim 14, wherein the dye is Fluorescein (Fluorescein), Eosin Y (Eosin Y), Eosin B (Eosin B), Erythrosine B (Phloxine B) , Group consisting of Rose Bengal, Merbromin, Ethyl Eosin, Rhodamine B, Rhodamine 6G and Sulforhodamine B Artificial photosynthesis method, characterized in that selected from. 15. The method of claim 14, wherein the sacrificial electron donor is triethanolamine (TEOA), ethylenediaminetetraacetic acid (EDTA), citric acid, formic acid, ascorbic acid, oxalic acid (Oxalic acid), an artificial photosynthesis method characterized in that it is selected from the group consisting of water and water. 15. The method of claim 14 wherein the electron mediator is selected from the group consisting of methylviologen, ruthenium II complex and rhodium III complex. 19. The method of claim 18 wherein the rhodium III complex is (pentamethylcyclopentadienyl-2,2'-bipyridinechloro) rhodium (III). The method of claim 14, wherein the redox enzyme is GDH (glutamate dehydrogenase), ADH (Alcohol dehydrogenase), G6PDH (glucose-6-phosphate dehydrogenase), LDH (lactic dehydrogenase), MDH (malate dehydrogenase) and SDH (succinic dehydrogenase) Artificial photosynthesis method, characterized in that selected from the group consisting of.

Images