TWI573875B - The use of a porphyrin compound used in the production method of a catalyst for lignin decomposition and a method for producing a tetrapyrrole compound - Google Patents

Description

Use of porphyrin compound used in a method for producing a catalyst for lignin decomposition and method for producing tetrapyrrole compound

The present invention relates to a catalyst for lignin decomposition, a method for producing an alcohol and an organic acid, a method for producing a lignin decomposition product, a catalyst for decomposing an aromatic hydrocarbon, a method for releasing hydrogen ions, and a porphyrin, particularly A catalyst for lignin decomposition by porphyrin, a method for producing an alcohol such as methanol from lignin, an organic acid, a method for producing a lignin decomposition product, a catalyst for decomposing an aromatic hydrocarbon by porphyrin, and a catalyst The lignin decomposition catalyst formed by porphyrin is a method of free hydrogen ions from lignin, and a porphyrin.

After entering the 21st century, the warming acceleration on the earth progressed, and carbon dioxide carbon became the control industry and even the key to controlling the world economy. As long as the fossil fuel blocked in the ground or on the sea floor is used as an energy source, it is impossible to reduce carbon dioxide in the atmosphere, and it is difficult to suppress the increase. In view of this, an alcohol such as bioethanol produced by plants is expected to be used as an energy source. However, at this time, in the past, technology used almost as a raw material for sugar, which caused a problem of competition between human food sources and energy. Recently, it has finally not competed with food sources, for example, alcohol technology has been advanced to use cellulose as a carbon source.

Grasses that cannot be wood or foodstuffs are mainly made up of cellulose and lignin. When the wood which is close to the so-called waste or the cellulose of the grass is used as a carbon source as an inappropriate material for waste materials and sheet materials, it is possible to suppress the carbon dioxide carbon emission and contribute to the industrial and economic circles.

For the use of cellulose as described above, the effective use of lignin, which is as rich as carbon, is still extremely limited. As a practical step, for example, it is simply burned as a heat source, or a preservative, or a structural strengthening agent mixed in cement.

Moreover, with the development of science and technology and industry, the accumulation of industrial waste that produces compounds that were originally present in non-high concentrations in nature has become a very large negative asset for human beings. Most of these compounds are compounds containing an aromatic hydrocarbon bonded to an oxygen atom on a carbon atom forming a benzene ring. For example, a dioxin class can be mentioned. Regarding the Dioxin class, although it has been considerably improved by the high temperature treatment of the waste burning stage of the source, there are many harmful compounds that have spread. Therefore, these harmful compounds are further decomposed by using sunlight which is originally injected into the natural environment to decompose the harmful compound, which can be decomposed by infinitely injected sunlight on the surface, which contributes to natural purification and human health maintenance. However, this useful decomposition catalyst has not yet been proposed.

Further, in recent years, as a source of energy, a fuel cell that receives an electromotive force by movement of hydrogen ions is attracting attention. However, when hydrogen is used as a source of hydrogen ions, hydrogen is almost always produced from fossil fuels. Further, hydrogen gas can be obtained by electrolysis of water, but power must be supplied at this time.

Further, when the power source is a solar cell obtained from sunlight, the manufacture of a semiconductor device is necessary. Therefore, if the current energy needs depend on the solar cell, resources and costs become a great burden. In the dye-sensitized solar cell, titanium oxide having a nanometer size is necessary, and a synthetic pigment having a certain electromotive force is expensive.

In the past, a decomposition method in which a lignin-based substance is brought into contact with functional water under light irradiation to decompose a lignin-based substance is known (for example, refer to Patent Document 1). This prior art describes sodium hydroxide as functional water, but it does not specifically describe the substance to be decomposed, and there is no description about obtaining an alcohol. Further, porphyrins containing a large number of Cl and F are known to be active in the reaction of acidification of lignin and conversion of an alkane to an alcohol (for example, refer to Patent Document 2). However, the use of an alkali compound or a photocatalyst has not been described in the prior art.

[Advanced technical literature]

[Patent Literature]

[Patent Document 1] JP-A-2000-144592 (Patent Application, Paragraph: 0030)

[Patent Document 2] Japanese Patent Publication No. 2-503086 (the upper left column of page 4)

An object of the present invention is to provide a method for producing an alcohol such as methanol and an organic acid, and a method for producing a lignin decomposition product, which is a lignin decomposition catalyst or lignin which is formed from a porphyrin which has not been used in the prior art. A catalyst for decomposing aromatic hydrocarbons derived from porphyrin, a method for using free ions of lignin as a raw material, and a porphyrin.

The catalyst for lignin decomposition of the present invention is characterized by containing a porphyrin exhibiting a catalytic function by light irradiation.

The catalyst for lignin decomposition of the present invention is further characterized by a porphyrin containing a catalyst function in an alkali solution.

The catalyst for lignin decomposition of the present invention is further characterized by a porphyrin containing a catalyst function by irradiation with light in an alkali solution.

In the above catalyst system for lignin decomposition, a tetrapyrrole compound containing (4) methyl groups and (4) ethyl esters or acetate groups (propionic acid groups) with porphyrin as (4) porphyrin rings is feature.

The catalyst system for lignin decomposition described above is characterized in that (four) porphyrins are tetrapyrrole compounds containing four methyl groups, four ethyl esters or acetoxy groups (propionic acid groups) in the porphyrin ring.

The above-mentioned catalyst system for lignin decomposition is characterized in that porphyrin is a porphyrin having a carboxyl group in a molecule.

The lignin decomposition catalyst system is characterized in that the porphyrin is a porphyrin having a total of two, four or eight carboxyl groups in the molecule.

The catalyst system for lignin decomposition described above is characterized in that porphyrin is at least one selected from the group consisting of urinary porphyrin, protoporphyrin, and coproporphyrin.

In the above-mentioned catalyst system for lignin decomposition, a porphyrin is a tetrapyrrole compound having a porphyrin ring structure obtained by culturing Escherichia coli in a culture medium.

The catalyst system for lignin decomposition described above is characterized in that the catalyst contains a medium containing a tetrapyrrole compound having a porphyrin ring structure obtained by culturing Escherichia coli in a culture medium.

The above-mentioned catalyst system for lignin decomposition is characterized by Escherichia coli which is incapable of expressing Escherichia coli as a gene ypjD (b2611).

For the above-mentioned catalyst system for lignin decomposition, Escherichia coli is used as a gene. The transposon of ypjD (b2611) is characterized by insertion of a mutant strain.

In the above catalyst system for lignin decomposition, the porphyrin is characterized in that no transition metal atom is coordinated at the center of the porphyrin ring.

The tetrapyrrole compound is produced by the Escherichia coli in which the gene ypjD (b2611) is rendered incapable of being expressed by mutation, or the expression function of the gene ypjD (b2611) is cultured in a medium in a partially or completely deficient state. The tetrapyrrole compound obtained from the medium is characterized.

The method for producing an alcohol and an organic acid according to the present invention is characterized in that a lignin is added to a solution containing an alkali compound, and an alcohol and an organic acid are separated from the lignin-base compound solution.

In the method for producing the above alcohols and organic acids, the alcohol is separated from the lignin-alkali compound solution by light irradiation (preferably by ultraviolet light or sunlight). Organic acids are characteristic.

In the method for producing the above alcohols and organic acids, the lignin-base compound solution is further subjected to the lignin decomposition catalyst, and the alcohol and the organic acid are separated from the lignin-base compound solution. feature.

In the method for producing the above-mentioned alcohols and organic acids, the lignin-base compound solution is further subjected to light irradiation (preferably by irradiation with ultraviolet rays or sunlight) after the lignin decomposition catalyst is acted on. The lignin-base compound solution is characterized by separating alcohols and organic acids.

The method for producing an alcohol and an organic acid according to the present invention is characterized in that an alcohol and an organic acid are separated from the lignin by the action of the lignin on the lignin decomposition catalyst.

The method for producing an alcohol and an organic acid according to the present invention is characterized in that an alcohol and an organic acid are separated from the lignin by light irradiation (preferably, irradiation with ultraviolet rays or sunlight).

In the method for producing the above-mentioned alcohols and organic acids, the lignin is decomposed by a catalyst for lignin, and then light-irradiated (preferably irradiated with ultraviolet rays or sunlight) to separate alcohols from lignin. And organic acids are characteristic.

The method for producing the above alcohols and organic acids is characterized in that the alkali compound is at least one of KOH and NaOH.

The method for producing the above alcohols and organic acids is characterized in that the alcohol is methanol and the organic acid is formic acid, acetic acid, malic acid, succinic acid or pyruvic acid.

The production method of the above alcohols and organic acids is characterized in that the separation of alcohols is carried out by distillation.

The method for producing a lignin decomposition product of the present invention is a method for producing a lignin decomposition product of a low molecular weight carbon-containing compound which is produced by separating an alcohol or an organic acid according to the method for producing an alcohol or an organic acid. Characterized.

The catalyst for decomposing an aromatic hydrocarbon which bonds an oxygen atom to a carbon atom forming a benzene ring of the present invention is characterized by containing a porphyrin.

The catalyst system for the decomposition of aromatic hydrocarbons is characterized in that the porphyrin is a tetrapyrrole compound having a porphyrin ring structure obtained by culturing Escherichia coli in a culture medium.

The catalyst system for the decomposition of aromatic hydrocarbons is characterized in that the catalyst is composed of a medium containing a tetrapyrrole compound having a porphyrin ring structure obtained by culturing Escherichia coli in a culture medium.

The above-mentioned catalyst for the decomposition of aromatic hydrocarbons is characterized by Escherichia coli which is expressed by mutation of Escherichia coli as a gene ypjD (b2611).

The above-mentioned aromatic hydrocarbon decomposition catalyst system is characterized in that a transposon (Transposon) insertion mutant strain of Escherichia coli is the gene ypjD (b2611).

The catalyst system for the decomposition of aromatic hydrocarbons is characterized in that the porphyrin is a tetrapyrrole compound containing a methyl group and an ethyl ester or an acetate group (propionic acid group) in the porphyrin ring.

The catalyst system for the decomposition of aromatic hydrocarbons is characterized in that the porphyrin is a tetrapyrrole compound containing four methyl groups, four ethyl esters or an acetate group (propionic acid group) in the porphyrin ring.

The catalyst system for the decomposition of aromatic hydrocarbons is characterized in that porphyrin is at least one selected from the group consisting of urinary porphyrin, protoporphyrin, coproporphyrin, and etioporphyrin.

The catalyst system for the decomposition of aromatic hydrocarbons is characterized in that the porphyrin is a porphyrin having a carboxyl group in the molecule.

The catalyst system for the decomposition of aromatic hydrocarbons is characterized in that porphyrin is a porphyrin having a total of two, four or eight carboxyl groups in the molecule.

The above-mentioned aromatic hydrocarbon decomposition catalyst system is characterized in that the aromatic hydrocarbon is a dioxin.

The free ion ion method of the present invention is characterized in that the lignin or lignin-alkali compound solution is subjected to the above-mentioned lignin decomposition catalyst, and the solution is subjected to light irradiation (preferably, irradiation with ultraviolet rays or sunlight). To make the hydrogen ion free.

The porphyrin system of the present invention is characterized by having a catalytic function of converting lignin into an alcohol and an organic acid.

The porphyrin of the present invention is further characterized by a catalyst function having a compound which decomposes an aromatic hydrocarbon having an oxygen atom bonded to a carbon which forms a benzene ring.

The porphyrin of the present invention is obtained by culturing the gene ypjD (b2611) by Escherichia coli which cannot be expressed by mutation.

The lignin decomposition catalyst of the present invention is further characterized by comprising a porphyrin obtained by culturing Escherichia coli which is incapable of expressing the gene ypjD (b2611) by mutation.

Effect of the invention

In the present invention, a porphyrin having a porphyrin structure or a synthetic porphyrin having a porphyrin structure produced by using Escherichia coli can be used as an active lignin decomposition catalyst and an aromatic hydrocarbon decomposition catalyst.

In the present invention, lignin is caused by a single or a combination of an alkali compound, light irradiation, and a catalyst for lignin decomposition, and an alcohol and an organic acid can be produced from lignin, and a decomposition product of a low molecular weight can be obtained, and The effect of free hydrogen ions can be achieved.

Form of implementing the invention

Hereinafter, embodiments of the present invention will be described in detail.

An embodiment of the catalyst for decomposing an aromatic hydrocarbon in which an oxygen atom is bonded to a carbon atom of a lignin and a benzene ring of the present invention, and the catalyst is contained by light irradiation and/or in an alkali solution. The porphyrin is a porphyrin having a catalytic function, and the porphyrin is preferably a tetrapyrrole compound or a synthetic porphyrin having a porphyrin ring structure in which Escherichia coli is cultured in a medium and secreted in a medium. For example, the catalyst may be a medium containing a tetrapyrrole compound having a porphyrin ring structure obtained by culturing Escherichia coli in a culture medium, or may be a cultured cell obtained by culturing or cultured. The extract obtained from the obtained cells can also be obtained.

The Escherichia coli in which the above-mentioned Escherichia coli is mutated, such as the gene ypjD (b2611), exhibits a change in the expression of Escherichia coli, and the transposon (Transposon) insertion mutant of the gene ypjD (b2611) is more good. The porphyrin is preferably a tetrapyrrole compound containing a methyl group (for example, four), an ethyl ester or an acetate group (propionic acid group) (for example, four) in the porphyrin ring.

The catalyst for decomposing lignin from Escherichia coli and the catalyst for decomposing aromatic hydrocarbons of the present invention can be produced, for example, by the following.

The medium for producing the above-mentioned catalyst is only a medium capable of culturing Escherichia coli, and can be used without particular limitation. Escherichia coli is cultured in a oligotrophic medium or a nutrient-rich medium, and the tetrapyrrole compound is separated and recovered from the medium to produce a porphyrin ring structure having a catalyst for lignin decomposition and a catalyst for decomposition of aromatic hydrocarbons. Pyrrole compound. In the process of culturing and proliferating Escherichia coli in a medium, a tetrapyrrole compound is produced in Escherichia coli, and a tetrapyrrole compound is produced by recovering a tetrapyrrole compound secreted in the medium. In order to prevent a component such as a natural substance contained in the medium from becoming a barrier to separation of the tetrapyrrole compound, it is preferred to use a poor nutrition medium, but the medium is not particularly limited. As the oligotrophic medium, those containing glucose or lactose are preferably used, but are not limited thereto.

It is preferable that the Escherichia coli used for the production of the catalyst for lignin decomposition and the catalyst for decomposing aromatic hydrocarbons is not mutated by the mutation of the gene ypjD (b2611). For example, Escherichia coli derived from the K12 strain and the BL21 strain can be mentioned. For example, it is preferred to use Escherichia coli which is not expressed by mutation from the gene ypjD (b2611) of the K12 strain. An Escherichia coli strain which is incapable of being expressed by mutation of the gene ypjD (b2611), for example, a transposon insertion mutant of the gene ypjD (b2611). This mutant is a state in which the expression of the gene ypjD (b2611) is partially or completely absent. Further, the K12 strain can be obtained, for example, from National BioResource, and the BL21 strain can be obtained, for example, from Takarabio. Further, as a transposon insertion mutant of the gene ypjD (b2611), for example, JD23504 and the like which are available from National BioResource.

For the embodiment of the present embodiment, Escherichia coli is first cultured in a oligotrophic medium. At this time, Escherichia coli is precultured in a medium other than a poor nutrient medium, for example, a medium such as an LB medium, and it is preferred that the obtained culture is inoculated into a oligotrophic medium to carry out the culture. Further, as the culture medium, a nutrient-rich medium other than the lean nutrient medium, a synthetic culture solution, or the like can be used. For example, a synthetic culture solution of an aqueous solution obtained by adding KH ₂ PO ₄ , K ₂ HPO ₄ , (NH ₄ ) ₂ SO ₄ , citric acid dihydrate, glucose, and MgSO ₄ may be used in the deionized water. It is only a medium which can increase Escherichia coli cells, and it is not limited to any of them.

The culture conditions of Escherichia coli are as follows for the general conditions of Escherichia coli. This is, for example, when the medium is cultured in the oligotrophic medium after the pre-culture of Escherichia coli, and the culture conditions are the same for any of the culture conditions. For example, using LB medium, culturing at a temperature of 15 ° C to 40 ° C for 6 hours to 24 hours, the resulting cell suspension is placed in a oligotrophic medium at a temperature of 20 ° C to 40 ° C for 12 hours to 96 hours. The cultivation of this. Thereby, the cells can be proliferated in the medium to obtain a culture (product) having a specific color tone of the tetrapyrrole compound.

Next, the intended tetrapyrrole compound was isolated from the above culture as follows.

Specifically, after filtering the supernatant obtained by centrifuging the culture, the tetrapyrrole compound is separated from the filtrate by, for example, an ion exchange resin column or a reverse phase column. For example, the culture is precipitated by a centrifugal separator to obtain a clear liquid containing the culture (product). Next, the clear liquid is filtered through a filter having a predetermined pore size (for example, 0.22 μm), and then adsorbed to the ion exchange resin using the column. Next, for example, the product is eluted from the ion exchange resin using a 20% acetonitrile-0.1% trifluoroacetic acid solution or the like, and then freeze-dried. And the dissolution at this time is such that it can be used for a solution containing an acid or a base in an organic solvent. In the embodiment of the present invention, one or two or more kinds of tetrapyrrole compounds can be obtained. For example, a milligram to several tens of mg of a tetrapyrrole compound can be obtained from 500 mL of a cell suspension.

When the product separated by the above operation was analyzed by NMR (Nuclear Magnetic Resonance) or the like, it was confirmed that the tetrapyrrole compound was contained. Further, when the product was analyzed by absorbance, it was found that the dye was a compound having absorption in a specific wavelength region. In most cases, a pigment compound exhibiting a bimodal peak similar to chlorophyll, protohemoglobin or ruthenium. Such a dye compound is useful as a photocatalyst or an electron transporter that excites electrons by light. Further, in the aqueous solution or in the cell membrane and the redox reaction, it is also conceivable that the battery can also function.

As described above, the use of Escherichia coli can produce a tetrapyrrole compound of a porphyrin such as a porphin or a porphyrin. Therefore, if it is a chemical synthesis method, it is necessary to prepare a device or a catalyst for the compound of the purpose, and it is not necessary. There are fewer concerns about the use of solvents that have a negative impact on the environment. Further, when Escherichia coli is cultured, a precursor of a tetrapyrrole compound such as 5-aminolevulinic acid is not required to be added to the culture medium (Japanese Laid-Open Patent Publication No. Hei 5-244937), and the tetrapyrrole compound is recovered and secreted in the culture medium, and it is not necessary to The bacterial cell is taken (Japanese Patent Publication No. Hei 5-91866). That is, since the culture of Escherichia coli or the recovery of the tetrapyrrole compound does not require a specific compound or device, the tetrapyrrole compound can be easily produced. The tetrapyrrole compound thus obtained can be used in various industrial fields such as medical, food, and electronics.

In the above, the tetrapyrrole compound is separated from the culture to serve as a catalyst for decomposition of lignin and a catalyst for decomposition of aromatic hydrocarbons. It is preferred to use a compound recovered from the culture, but the culture or the cells obtained by the culture may contain the tetrapyrrole compound, and the culture may be used as a catalyst for lignin decomposition or a catalyst for decomposition of aromatic hydrocarbons. use.

The catalyst for lignin decomposition and the catalyst for decomposing aromatic hydrocarbons which can be used in the present invention, as the other synthetic porphyrin, for example, may be selected from the group consisting of protoporphyrin, urinary porphyrin, coproporphyrin, and primary purple. At least one kind of quality. However, aetioporphyrin cannot decompose lignin. As the porphyrin, in the following examples, protoporphyrin IX (manufactured by ALDRICH Co., Ltd.) having two carboxyl groups in the molecule, urinary porphyrin I (manufactured by SIGMA) having eight carboxyl groups in the molecule, and four molecules in the molecule were used. Carbium porphyrin I (manufactured by ALDRICH Co., Ltd.) and primula (manufactured by ALDRICH Co., Ltd.) which does not contain a carboxyl group in the molecule. Similarly to the porphyrin obtained by culturing the above Escherichia coli, the porphyrin does not have a transition metal atom coordination at the center of the porphyrin ring.

Next, an embodiment of a method for producing an alcohol and an organic acid according to the present invention will be described. The alcohol and the organic acid in the present invention can be produced by using lignin alone or in combination with an alkali compound, light irradiation, or the above-mentioned catalyst for lignin decomposition.

In other words, the alcohol such as methanol is, for example, (1) a solution containing at least one alkali compound selected from the group consisting of KOH and NaOH, and the lignin-base compound solution is separated from the alcohol by distillation or the like, for example. 2) A solution containing lignin added to the above-mentioned alkali compound, and ultraviolet rays are used as the lignin-base compound solution (for example, an ENF type (260c/j, or 280c/j, etc.) or UVP manufactured by Spectronix Co., Ltd. can be used. After the light having a wide wavelength region such as ultraviolet rays of UVL-56Hand Held or the like, which is irradiated by the company, is irradiated for a predetermined period of time, the lignin-alkali compound solution after irradiation can be efficiently used, for example, by distillation or the like. Separating the alcohol, (3) adding the lignin solution containing the alkali compound, and applying the lignin decomposition catalyst to the lignin-base compound solution at a predetermined temperature for a predetermined period of time, and then applying the solution For example, the alcohol can be efficiently separated by distillation, (4) the solution containing the above-mentioned alkali compound added to the lignin, and the catalyst for decomposing the above lignin is used for the lignin-base compound solution. After the predetermined time is applied at a temperature, the solution is irradiated with light having a wide wavelength region such as ultraviolet light or sunlight for a predetermined hour, and secondly, the lignin-alkali compound solution after the irradiation is efficiently distilled, for example, or the like. The alcohol is separated, and (5) for the lignin, the lignin decomposition catalyst is allowed to act at a predetermined temperature for a predetermined period of time, and the solution can be efficiently separated, for example, by distillation or the like, and (6) The lignin is irradiated with light having a wide wavelength region such as ultraviolet light or sunlight for a predetermined period of time, and secondly, the solution can efficiently separate the alcohol, for example, by distillation, and (7) the lignin is lignin for the lignin After the decomposition catalyst is subjected to a predetermined time at a predetermined temperature, the solution is irradiated with light having a wide wavelength region such as ultraviolet rays or sunlight for a predetermined period of time, and secondly, the irradiated lignin-alkali compound solution is used, for example, by It can be produced by efficiently separating alcohols by distillation or the like. When the light irradiation is carried out, when the reaction liquid containing lignin is brought into contact with a gas containing air or oxygen, oxygen and/or nitrogen, and the like, the catalyst function of the porphyrin can be efficiently exhibited. . For example, when the lignin concentration is 2.5 mg/mL and the porphyrin is 50 μg/mL (mass ratio 50:1), oxygen of about 0.2 to 0.5 mL per lignin can be used.

Further, organic acids such as formic acid, acetic acid, malic acid, succinic acid, and pyruvic acid are the same as those of the above alcohols, and can be separated from lignin.

The lignin to be used in the present invention is not particularly limited, and for example, a product having a high purity which does not contain impurities such as reducing sugar or cellulose (for example, catalog No. 471003 manufactured by SIGMA Co., Ltd., molecular weight 60,000; catalogue manufactured by ALDRICH Co., Ltd., can be used. No. 471,046, molecular weight: 12,000), a product having a slightly lower purity containing reducing sugar (for example, catalog No. 471038 manufactured by SIGMA Co., Ltd., molecular weight: 52,000), and a water-insoluble product (for example, catalog No. 370967 manufactured by ALDRICH Co., Ltd.) . In the present invention, an alcohol such as methanol and an organic acid such as formic acid, acetic acid, malic acid, succinic acid or pyruvic acid can be isolated from all of these lignin. That is, irrespective of the presence of impurities, the average molecular weight, and solubility in water, only lignin can be produced by separating alcohols and organic acids by the present invention.

The solution of the alkali compound is not particularly limited, and for example, KOH and/or NaOH of about 0.0025 M to 0.05 M can be used. There is a high level of separation efficiency between the alcohol and the organic acid, and the solution of the alkali compound is not limited in the concentration range.

In one embodiment of the catalyst for decomposing an aromatic hydrocarbon which bonds an oxygen atom to a carbon atom of the benzene ring of the present invention, the catalyst has the same composition as the catalyst for lignin decomposition which is formed by the porphyrin. The detailed description is omitted.

Examples of the aromatic hydrocarbon that causes the catalyst to act include dioxin or dioxin-like compounds. The dioxin includes, for example, polychlorinated diphenyl dioxide (PCDD), polychlorinated dibenzofuran (PCDF), and the like, and a dioxin-like compound contains, for example, polychlorinated biphenyl (Dioxin-like PCB). The catalyst for decomposing aromatic hydrocarbons of the present invention is a non-toxic person obtained by decomposing these dioxins.

Further, another embodiment of the present invention is a method in which a lignin-free compound ion is added to a lignin-base compound solution containing a pyrrole compound derived from the above-mentioned Escherichia coli to form a porphyrin or a synthetic porphyrin. The lignin is decomposed with a catalyst, and at the same time as the free hydrogen ions, the light decomposition of lignin can be performed.

Further, in another embodiment of the present invention, according to the method for producing an alcohol or an organic acid, a compound containing a low molecular weight carbon which is a lignin decomposition product produced when an alcohol or an organic acid is separated can be recovered.

The present invention is not limited to the above-described embodiments, and various modifications can be made without departing from the spirit and scope of the invention. The manufacturing examples and examples of the present invention will be described in detail below.

(Manufacturing Example 1)

As described above, by adding porphyrin (pyrrole compound) to lignin as a catalyst for lignin decomposition, alcohols and organic acids can be produced, and photodecomposition of lignin can be carried out to obtain a decomposition product, and free hydrogen can be obtained. ion. Further, a tetrapyrrole compound or a synthetic porphyrin having a porphyrin structure is effective as a catalyst for decomposition of an aromatic hydrocarbon such as dioxin. In this case, as the porphyrin, for example, a biological method using Escherichia coli can be used. In the production example, a production example using an azole compound of Escherichia coli will be described.

First, a transposon (Transposon) derived from Escherichia coli gene ypjD (b2611) was inserted into a cell of a mutant strain (National BioResource JD23504) in 2 mL of LB medium (Bacto-tryptone: 1%, Bacto-yeast extract: 0.5%, In the NaCl: 0.5%), pre-culture was carried out at 37 ° C for 12 hours. 1 mL of the obtained cell suspension was added to 1 L of deionized water, KH ₂ PO ₄ was 9 g, K ₂ HPO ₄ was 21 g, (NH ₄ ) ₂ SO ₄ was 2 g, citric acid dihydrate was 1 g, and glucose was 3.6 g and 500 mL of an aqueous solution of 200 mg of MgSO ₄ were used, and the main culture was carried out at 37 ° C for 24 hours.

The color of the obtained culture solution was such that it was colorless at the time of starting the main culture, but became pink after 24 hours. The culture solution was centrifuged to precipitate cells, and the resulting clear liquid was filtered through a filter having a pore size of 0.22 μm. Next, after the filtrate was filled through the column with an anion exchange resin, the culture absorbing the resin was dissolved in a 20% acetonitrile-0.1% trifluoroacetic acid solution, and the lysate was freeze-dried. Thereby, a product with a pink color is obtained.

This product was confirmed to have a tetrapyrrole compound having the structure shown in Fig. 1 when subjected to various machine analyses described below. Symbols A to G in Fig. 1 are symbols corresponding to the ¹³ C NMR spectrum shown in Fig. 4, and are the same for the other planes. Further, when the product was analyzed by NMR, it was confirmed that the tetrapyrrole compound was contained. Further, potassium (K) was detected by ICP (Inductively Coupled Plasma) mass spectrometry, and recovered as a compound which forms K and ion-bound or erroneous. Further, when the spectrophotometric analysis was carried out, as shown in Fig. 2, the porphyrin-specific bimodal peak containing the Soret band was confirmed. In Fig. 2, there are peaks at wavelengths of 395 nm and 549 nm. From this, it was confirmed that the product was a free electron-moving organic pigment.

The tetrapyrrole compound obtained above was dissolved in a dissimilar solvent (Sample 1 and Sample 2), and subjected to secondary NMR measurement (COSY, NOESY, HSQC, HMBC), and the spectrum was analyzed and subjected to structural analysis.

In the sample 1, the sample was dissolved in CD ₃ OD, and NMR measurement was carried out under the following conditions.

‧Device: INOVA500 (variance company)

‧Resonance frequency: 499.8MHz ( ¹ H)

‧Base: 3.31 ppm (CD ₂ HOD, ¹ H NMR) 49.421 ppm (CD ₃ OD, ¹³ C NMR)

‧ Cumulative number: ¹ H NMR (16 times), ¹³ C NMR (53428), COSY (16 times), NOESY (8 times), HSQC (32 times), HMBC (128 times)

‧Other: The mixing time of NOESY is set to 400msec.

Further, in the sample 2, the sample was dissolved in a solvent of CD ₃ CN:D ₂ O:CD ₃ COOD=90:10:0.1, and NMR measurement was carried out under the following conditions.

‧Device: INOVA600 type (varian company)

‧Resonance frequency: 599.8MHz ( ¹ H)

‧Base: 1.92 ppm (CD ₂ HCN, ¹ H NMR) 1.28 ppm (CD ₃ CN, ¹³ C NMR)

‧ Cumulative times: ¹ H NMR (64 times), ¹³ C NMR (50000), COSY (16 times), NOESY (16 times), HSQC (32 times), HMBC (128 times)

‧Other: The mixing time of NOESY is set to 400msec.

Structural analysis of sample 1:

Figure 3 shows the ¹ H NMR spectrum (solvent: CD ₃ OD). As a result, a signal estimated to be from the target component was observed in the vicinity of 10.0 to 10.5 ppm (d), in the vicinity of 4.3 ppm (f), in the vicinity of 3.6 ppm (g), and in the vicinity of 3.2 ppm (e). The signal strength ratio is approximately 1:2:3:2. The d~g are the same on other maps.

Figure 4 shows the ¹³ C NMR spectrum. It is presumed to be the aromatic of the signal from B and C.

Among them, the ¹ H NMR d signal is generally a characteristic signal that cannot be seen from the chemical shift value, and may be a porphyrin skeleton if considered as an aromatic compound. As described below, when the binary NMR spectrum (Figs. 5 to 9) is analyzed as a porphyrin, it can be analyzed without contradiction. Further, a compound similar to the putative structure of Fig. 1 has been reported in J. org. Chem. Vol. 164, No. 21, 1999 (7973-7982), and the chemical shift values of ¹ H NMR are shown to be quite uniform, so it is presumed to be The porphyrin structure is more appropriate.

The analysis of the binary NMR will be described below.

Figure 5 shows the HSQC spectrum. The HSQC spectrum is an assay for detecting J ¹ _CH . The analysis results are shown in the figure. For the proton signal, the number is recorded on the large text of the direct-bonded carbon number.

Figure 6 shows the COSY spectrum. The COSY spectrum is an assay for detecting spin coupling between ¹ H- ¹ H. The analysis results show that f and e are spin couplings, and it is highly probable that the signal intensity ratio and the chemical shift values of F and E are -CH ₂ -CH ₂ -X. Wherein X represents an undetermined component of unknown structure.

Figure 7 shows the HMBC spectrum. The HMBC spectrum is obtained by measuring the J ⁿ _CH (n=2 to 4 degree) to obtain a heterogeneous nuclear distant binding-associated spectrum. The spectrum is analyzed to obtain (e, A), (e, B), (e, F), (g, B), (g, C), (f, A), (f, B), ( f, C), (f, E), etc. These correlations are not inconsistent with the putative structure shown in Figure 1.

Figures 8 and 9 show the NOESY spectrum. The NOESY spectrum is a measurement method for detecting magnetization exchange, and the distance between the nuclear spins can be obtained by the magnetization movement by the gradual relaxation. As a result of the analysis of the spectrum, NOE correlation was observed at (g, e), (f, g), (f, e), (d, f), (d, e), (d, g). These NOE correlations are those that support the putative structure shown in FIG.

Sample 2 related structure analysis:

Fig. 10 shows a ¹ H NMR spectrum, and Fig. 11 shows a ¹³ C NMR spectrum (solvent: CD ₃ CN: D ₂ O: CD ₃ COOD = 90: 10: 0.1). The signal surrounded by the square frame in Fig. 10 indicates impurities. As a result of comparison with the spectrum of the above sample 1, it was estimated that the structures of the main components were the same. This can also be supported by the analysis of the spectra of COSY, NOESY, HSQC, and HMBC.

Fig. 12 shows the NOESY spectrum of the expanded sample 2. Since four kinds of d protons were observed to be separated, the numbers from d1 to d4 were given from the low magnetic field (the smaller number). Inductive NOE related as follows.

‧ d1 and d4 are simultaneously associated with methyl and -CH ₂ -CH ₂ -X for NOE.

‧ d2 is only associated with -CH ₂ -CH ₂ -X for NOE.

‧ d3 is only related to methylation as NOE.

This is related to the NOE in which the methyl groups are not specifically observed or -CH ₂ -CH ₂ -X are related to each other, and the side chain configuration of Fig. 1 is obtained.

The numbers of the ¹³ C signal and the ¹ H signal are assigned a number for signals observed in almost the same area. The reason for this is that the skeleton of the porphyrin is a repetitive structure, so it is difficult to classify it in detail. Regarding the number of repetitions, four methyl groups (g) were observed, so four were estimated.

As described above, from the analysis results of the samples 1 and 2, the structure of Fig. 1 was obtained. However, the total of the methyl group of the side chain and -CH ₂ -CH ₂ -X may be four, and the additional portion is not in any of the eight places shown in FIG. 1 and is not limited to the data. Show additional location.

Next, a review is made for the X portion corresponding to -CH ₂ -CH ₂ -X.

The following analysis was carried out for the product obtained above.

(1) Qualitative analysis by thermal decomposition GC/MS:

After removing the TFA or the like, the sample is heated in a heating furnace at 280 ° C for 10 minutes, and then thermally decomposed at 600 ° C to form a decomposition product in the gas chromatography layer/mass analyzer described below (hereinafter referred to as "GC/MS"). ) Perform separation analysis.

Device name: HP5973 model manufactured by Agilent Technologies with quality selective detector

HP6890 gas color layer

FRONTIER LAB PY-2020iD heating furnace type thermal decomposition device

As for the measurement sample, a solution of 1 mL of AcCN was added to 2 mL of the sample, and 0.5 mL of the solution was poured into the sample cup, and then AcCN was removed by rinsing with nitrogen.

(2) Analysis by electrospray-mass analysis device:

The molecular weight of the component in the solution was examined and analyzed by an electric spray-mass analyzer (hereinafter referred to as "ESI-MS") described below.

Device name: Qstar made by Applied Biosystems

Introduced solvent: AcCN/0.05% formic acid in water (50/50)

Introduction method: The measurement solution was directly introduced using a 30 μL loop wire.

Measurement mode: Positive mode

The solution was measured: 2 mg of the solution of AcCN was added to a solution of 1 mL of AcCN in a small glass vial, and diluted to about 100 ppm with an introduction solvent.

Carbon dioxide carbon was detected from the sample by the analysis of the above GC/MS. Further, as a result of the analysis by the above ESI-MS (Fig. 14), fragment ions having a molecular weight of 59 (four peaks) were detected, and it was confirmed that the fragment contained an ethyl ester or an acetate group (propionic acid group).

Further, the product was analyzed by thermal decomposition GC-MS, and as shown in Fig. 15, the main component was a pyrrole compound. This can be confirmed by comparing the mass spectrum of each component with the data base value, and the structure containing the pyrrole ring in the molecule can be confirmed. Further, as for the detailed molecular weight, as shown in Fig. 13, ions of additional hydrogen ions were detected to obtain a molecular weight of 655.2760-1.0078 = 654.2682. From the above, it was confirmed that the side chain contains four ethyl esters or an acetate group (propionic acid group), the molecular weight of four methyl groups is 654, and the porphyrin compound of the formula C ₃₆ H ₃₈ O ₈ N ₄ is confirmed to be a porphyrin ring. There is no transition metal in the center.

[Example 1]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of a solution containing the lignin: 2.5 mg/mL, 0.05 M KOH, and porphyrin having a carboxyl group of 50 μg/mL in the molecule obtained in the above Production Example 1 was placed in an Eppendorf tube having a polypropylene degree. The light-transmissive cylindrical test tube was irradiated with ultraviolet light for 12 hours using an ENF-type ultraviolet lamp manufactured by Spectronix. Thereafter, the mixture was heated at 80 ° C for 60 minutes, and the volatilized gas was analyzed using Shimadzu GC/MS QP-2010, column DB-WAX, and methanol of 170 μg/mL was measured. At this time, 140 μg/mL of formic acid, 25 μg/mL of malic acid, 19 μg/mL of acetic acid, 5.4 μg/mL of succinic acid, and 6.2 μg/mL of pyruvic acid were obtained. At this time, methanol equivalent to 6.8% by weight, equivalent to 5.6% by weight of formic acid, was obtained on the basis of the weight of dry lignin.

In addition, as a lignin, a product having a high purity (a product of No. 471,046, manufactured by Aldrich Co., Ltd., molecular weight: 12,000), and a product having a slightly lower purity (available from Sigmal Co., Ltd., Catalog No.) is used as a lignin. .471038, a molecular weight of 52,000), and a product insoluble in water (Aldrich, catalog No. 370967), after repeating the above process, the same amount of methanol and the amount of the above organic acid were obtained. That is, irrespective of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water, it is known that alcohols and organic acids can be separated from lignin.

Further, when sunlight is used instead of ultraviolet light, the same results as the amount of methanol and the amount of organic acid are obtained after repeating the above process.

A sample of the reaction liquid obtained by irradiation with ultraviolet light as described above was subjected to HPLC (High Speed Liquid Chromatography) and analyzed using a colloidal filter column. As a control, the solution before ultraviolet irradiation was also analyzed. As a result of the analysis, the lignin was exposed to light at 310 nm before irradiation, and the peak of dissolution at 7-9 min was detected, but as described above, when porphyrin was added to lignin and converted to methanol or organic acid by ultraviolet light irradiation, 310 nm was used. In the light absorption, the peak area of the dissolved lignin at 7-9 min was reduced to 20%. That is, it is known that when porphyrin is added to lignin and irradiated with ultraviolet light, 80% of lignin is decomposed. The decomposition product is filtered by colloid to dissolve off the lower molecular weight fraction. Therefore, about 8.5% by weight of methanol and about 7% by weight of formic acid can be obtained on the basis of the decomposed lignin.

[Embodiment 2]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. The lignin having a lignin: 2.5 mg/mL, 0.05 M KOH, and a protoporphyrin IX (manufactured by ALDRICH Co., Ltd.) containing two carboxyl groups in a molecule of a synthetic porphyrin, and a urinary porphyrin I having eight carboxyl groups in the molecule 1 mL of each 50 μg/mL solution containing 4 carboxyl groups of fecal porphyrin I (manufactured by ALDRICH Co., Ltd.) in a molecule, and placed in a light-transmissive cylindrical test tube having a degree of Eppendorf tube made of polypropylene. It was irradiated with ultraviolet light for 12 hours using an ENF type ultraviolet lamp manufactured by Spectronix. The obtained reaction liquid was heated at 80 ° C for 60 minutes, and the volatilized gas was analyzed using Shimadzu GC/MS QP-2010, column DB-WAX. After repeating the above operation several times, methanol and formic acid which were almost the same amount as in the case of Example 1 were obtained on the basis of the weight of dry lignin based on the conditions of the original porphyrin IX, urinary porphyrin I, and coproporphyrin I. That is, about 6 to 9% by weight, based on the weight of dry lignin, is converted to methanol, and about 2 to 4% by weight is converted to formic acid.

A sample of each reaction liquid obtained as described above was analyzed by HPLC (High Speed Liquid Chromatography) using a colloidal filter column. As a control, the solution before ultraviolet irradiation was also analyzed. As a result of the analysis, before the irradiation, lignin was detected to absorb peaks at 7-9 min in the light absorption at 310 nm, but the above porphyrins were added to lignin, and ultraviolet light was converted into alcohols and organic acids (formic acid, etc.). In the reaction solution, the protoporphyrin IX was reduced to about 60%, the urinary porphyrin I was reduced to about 15%, and the coproporphyrin I was reduced to about 20%. That is, it is known that when porphyrin is added to lignin and irradiated with ultraviolet light, the protoporphyrin IX is decomposed by about 40%, the urinary porphyrin I is decomposed by about 85%, and the coproporphyrin I is decomposed by about 80% lignin. The decomposition product is soluble in the lower molecular weight fraction by colloidal filtration.

In addition, as a lignin, a product having a high purity (a product of No. 471,046, manufactured by Aldrich Co., Ltd., molecular weight: 12,000) and a product having a slightly lower purity (available from Sigmal Co., Ltd.) is used as a lignin. .471038, molecular weight 52,000), and a product insoluble in water (Aldrich, catalog No. 370967), after repeating the above process, the use of synthetic porphyrin also gave the same degree of decomposition of lignin. That is, regardless of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water, the same degree of lignin can be decomposed.

Further, when sunlight is used instead of ultraviolet light, the same result as the decomposition of lignin can be obtained after repeating the above process.

[Example 3]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of a solution containing the lignin: 1.8 mg/mL, 0.05 M KOH, and porphyrin IV (Cat. No. 258535-1G) manufactured by Sigmal Co., Ltd.: 50 μg/mL, and placed in a polypropylene-made Eppendorf tube The light-transmissive cylindrical test tube was irradiated with ultraviolet light for 12 hours using an ENF-type ultraviolet lamp manufactured by Spectronix. Thereafter, the mixture was heated at 80 ° C for 60 minutes, and when volatilized gas was analyzed using Shimadzu GC/MS QP-2010 and column DB-WAX, 54 μg/mL of methanol was measured. That is, 3.0% by weight of methanol can be obtained on the basis of the weight of dry lignin.

In addition, as a lignin, a product having a high purity (a product of No. 471,046, manufactured by Aldrich Co., Ltd., molecular weight: 12,000) and a product having a slightly lower purity (available from Sigmal Co., Ltd.) is used as a lignin. .471038, molecular weight 52,000), and a product insoluble in water (Aldrich, catalog No. 370967), after repeating the above process, the same degree of methanol was obtained. That is, it was found that alcohol was separated from lignin by the presence of impurities in the lignin to be used, the average molecular weight of lignin, or the solubility of lignin in water.

Further, when sunlight was used instead of ultraviolet light, the same results as the amount of methanol were obtained after repeating the above process.

[Example 4]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of a solution containing the lignin: 7.5 mg/mL and 0.05 M KOH was placed in a light-transmissive cylindrical test tube having a degree of Eppendorf tube made of polypropylene, and heated at 80 ° C for 60 minutes to volatilize the gas. After analysis by Shimadzu GC/MS QP-2010 and column DB-WAX, 93 μg/mL of methanol was measured. That is, 1.24% by weight of methanol can be obtained on the basis of the weight of dry lignin.

In addition, as a lignin, a product having high purity such as reducing sugar or cellulose, a product having high purity (Aldrich, catalog No. 471,046, molecular weight: 12,000), and a product insoluble in water (Aldrich, catalog No. 370967) are used. After repeating the above process, the same amount of methanol was obtained. That is, it was found that methanol of the same degree can be produced from lignin regardless of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water.

[Example 5]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of a solution containing the lignin: 7.5 mg/mL and 0.05 M KOH was placed in a light-transmissive cylindrical test tube having a degree of Eppendorf tube made of polypropylene, and an ultraviolet light of ENF type by Spectronix was used for ultraviolet light. Light was irradiated for 24 hours. Thereafter, the mixture was heated at 80 ° C for 60 minutes, and when the volatile gas was analyzed using Shimadzu GC/MS QP-2010 and column DB-WAX, 150 μg/mL of methanol was measured. That is, 2% by weight of methanol based on the weight of dry lignin can be obtained.

In addition, as a lignin, a product having high purity such as reducing sugar or cellulose, a product having high purity (Aldrich, catalog No. 471,046, molecular weight: 12,000), and a product insoluble in water (Aldrich, catalog No. 370967) are used. After repeating the above process, the same amount of methanol was obtained. That is, it was found that the same degree of methanol was produced from lignin regardless of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water.

Further, when sunlight was used instead of ultraviolet light, the same results as the amount of methanol were obtained after repeating the above process.

[Embodiment 6]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of a solution containing lignin: 2.5 mg/mL, 0.05 M KOH, and porphyrin: 25 μg/mL obtained in the above Production Example 1 was placed in a light-transmitting cylinder having a degree of Eppendorf tube made of polypropylene. The test tube was irradiated with ultraviolet light for 12 hours using an ENF type ultraviolet lamp manufactured by Spectronix. Thereafter, the mixture was heated at 80 ° C for 60 minutes, and the volatilized gas was analyzed using Shimadzu GC/MS QP-2010 and column DB-WAX to measure 160 μg/mL of methanol. That is, 6.4% by weight of methanol can be obtained on the basis of the weight of dry lignin. It was found that by further adding a porphyrin as a photocatalyst to the lignin-base compound solution, the amount of methanol produced from lignin can be dramatically promoted.

In addition, as a lignin, a product having high purity such as reducing sugar or cellulose, a product having high purity (Aldrich, catalog No. 471,046, molecular weight: 12,000), and a product insoluble in water (Aldrich, catalog No. 370967) are used. After repeating the above process, the same amount of methanol was obtained. That is, it was found that methanol of the same degree can be produced from lignin regardless of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water.

Further, when sunlight was used instead of ultraviolet light, the same result was obtained as the amount of methanol after repeating the above process.

[Embodiment 7]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of the solution containing the lignin: 2.5 mg/mL was placed in a light-transmissive cylindrical test tube having a degree of Eppendorf tube made of polypropylene, and ultraviolet light was irradiated for 24 hours using an ENF-type ultraviolet lamp manufactured by Spectronix. Thereafter, the mixture was heated at 80 ° C for 60 minutes, and when the volatile gas was analyzed using Shimadzu GC/MS QP-2010 and column DB-WAX, 21 μg/mL of methanol was measured. That is, 0.84% by weight of methanol can be obtained on the basis of the weight of dry lignin.

In addition, as a lignin, a product having high purity such as reducing sugar or cellulose, a product having high purity (Aldrich, catalog No. 471,046, molecular weight: 12,000), and a product insoluble in water (Aldrich, catalog No. 370967) are used. After repeating the above process, the same amount of methanol was obtained. That is, it was found that the same degree of methanol was obtained from lignin regardless of the presence of impurities in the lignin used, the average molecular weight of lignin, and the solubility of lignin to water.

Further, when sunlight is used instead of ultraviolet light, the same result is obtained for the amount of methanol after repeating the above process.

[Embodiment 8]

In the present embodiment, the decomposition treatment method of the aromatic hydrocarbon decomposition catalyst of the present invention will be described with respect to the aromatic hydrocarbon to which the oxygen atom is bonded to the benzene ring.

Remazol Brilliant Blue (RBBR; manufactured by SIGMA Corporation) containing a compound containing an aromatic hydrocarbon bonded to an oxygen atom in a benzene ring was used for photolysis. The RBBR is a similar test drug used as an indicator for decomposing dioxin.

RBBR: 250 μg/mL, 0.05 M KOH, and porphyrin, protoporphyrin IX (manufactured by ALDRICH), urinary porphyrin I (manufactured by SIGMA), and porphyrin I (ALDRICH) obtained in accordance with the above Production Example 1. 1 mL of each 25 μg/mL solution (30% acetonitrile, 0.1% trifluoroacetic acid, 60% methanol as a solvent) was placed in a light-transmissive cylindrical test tube of a degree of Eppendorf tube made of polypropylene. The light was irradiated with ultraviolet light for 2 hours using an ENF type ultraviolet lamp manufactured by Spectronix. As a result, when any of the porphyrins and the solvent were used, it was found that the blue tone of the RBBR disappeared and the RBBR was decomposed.

Further, when sunlight is used instead of ultraviolet light, the decomposition of RBBR can be confirmed by repeating the above process.

[Embodiment 9]

In the present example, the photodecomposition was carried out using cyanoxylene (manufactured by Takara Co., Ltd.) and bromophenol blue (manufactured by Takara Co., Ltd.) containing a compound having an aromatic hydrocarbon bonded to an oxygen atom on a benzene ring.

The mixture contains cyanoxylene: 5 mg/mL, bromophenol blue: 5 mg/mL, 0.05 M KOH, and the porphyrin, protoporphyrin IX (manufactured by ALDRICH Co., Ltd.), and urinary porphyrin I (SIGMA) obtained according to the above Production Example 1. A solution of 40 μg/mL of each of the coumaline I (manufactured by ALDRICH Co., Ltd.) and primrose (manufactured by ALDRICH Co., Ltd.) (using 30% acetonitrile and 0.1% trifluoroacetic acid as a solvent) was placed in a holding polymer. A light-transmissive cylindrical test tube of a propylene-made Eppendorf tube was irradiated with ultraviolet light for 2 hours using an ENF-type ultraviolet lamp manufactured by Spectronix. As a result, when any of the porphyrins and the solvent were used, the blue and yellow hues of cyanoxylene and bromophenol blue disappeared, and it was found that cyanoxylene and bromophenol blue were decomposed.

Further, when sunlight is used instead of ultraviolet light, the decomposition of cyanoxylene and bromophenol blue can be confirmed by repeating the above process.

[Embodiment 10]

In the present embodiment, the lignin-decomposing catalyst of the present invention is used as a lignin in a method of using lignin free hydrogen ions as a lignin, and a product having high purity (such as Sigmal Co., Ltd., catalog No.) .471003, molecular weight 60,000).

1 mL of a solution containing the lignin 2.5 mg/mL and the porphyrin: 50 μg/mL obtained in the above Production Example 1 was placed in a light-transmissive cylindrical test tube having a degree of Eppendorf tube made of polypropylene, and Spectronix was used. The company's ENF UV lamp is exposed to UV light for 12 hours. As a result, the pH of the lignin reaction solution was lowered from 9.2 to 6.4. Therefore, it is known that free hydrogen ions can be derived from lignin.

In addition, as a lignin, a product having a high purity (a product of No. 471,046, manufactured by Aldrich Co., Ltd., molecular weight: 12,000) and a product having a slightly lower purity (available from Sigmal Co., Ltd.) is used as a lignin. .471038, molecular weight 52,000), and a product insoluble in water (Aldrich, catalog No. 370967), after repeating the above process, hydrogen ions were similarly released. That is, it is known that irrespective of the presence of impurities in the lignin to be used, the average molecular weight of lignin, or the solubility of lignin in water, hydrogen ions can be released to the same extent by lignin.

Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained after repeating the above process.

[Example 11]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of a solution containing lignin: 2.5 mg/mL, 2.5 mM KOH, and porphyrin: 50 μg/mL obtained in the above Production Example 1 was placed in a light-transmitting cylinder having a degree of Eppendorf tube made of polypropylene. The test tube was irradiated with ultraviolet light for 12 hours using an ENF type ultraviolet lamp manufactured by Spectronix. The sample was analyzed by HPLC (High Speed Liquid Chromatography) using a colloidal filter column. As a control, the solution before ultraviolet irradiation was also analyzed. As a result of the analysis, the peak dissolved in 7-9 min was detected in the light absorption at a wavelength of 310 nm before the irradiation, but the porphyrin was added to the lignin, and when irradiated with ultraviolet light, in the absorption at 310 nm, at 7-9 min. The peak area of the dissolved lignin was reduced to 78%. That is, it was found that when porphyrin was added to lignin, when irradiated with ultraviolet light, 22% of the mass was decomposed. Further, the pH of the lignin reaction solution was lowered from 10.7 to 6.2. Therefore, free hydrogen ions from lignin are known.

In addition, as a lignin, a product having a high purity (a product of No. 471,046, manufactured by Aldrich Co., Ltd., molecular weight: 12,000) and a product having a slightly lower purity (available from Sigmal Co., Ltd.) is used as a lignin. .471038, molecular weight 52,000), and a product insoluble in water (Aldrich, catalog No. 370967), the same results were obtained after repeating the above process. That is, it is known that, regardless of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water, the lignin is decomposed to the same extent, and hydrogen ions can be simultaneously treated to the same extent. free.

Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained after repeating the above process.

[Embodiment 12]

In the method of the lignin-decomposing catalyst of the present invention, a product having a slightly lower purity (manufactured by Sigmal Co., Ltd., catalog No. 471038, molecular weight: 52,000) is used as the lignin in the method of using lignin free hydrogen ions.

1 mL of a solution containing 5 mg/mL of the above lignin and 50 μg/mL of the porphyrin compound obtained in the above Production Example 1 was placed in a light-transmissive cylindrical test tube having a degree of Eppendorf tube made of polypropylene, and manufactured by Spectronix Co., Ltd. The ENF type ultraviolet lamp was irradiated with ultraviolet light for 12 hours. As a result, the pH of the lignin reaction solution was lowered from 6.2 to 4.8. Therefore, it is known that free hydrogen ions are derived from lignin.

In addition, as a lignin, a product having a high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000, and Aldrich, catalog No. 471046, molecular weight 12,000) is used, and impurities are not dissolved. The same result was obtained after repeating the above process in a product of water (made by Aldrich Co., catalog No. 370967). That is, it is known that irrespective of the presence of impurities in the lignin to be used, the average molecular weight of lignin, or the solubility of lignin in water, hydrogen ions can be released to the same extent by lignin.

Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained after repeating the above process.

[Example 13]

In the method of using a lignin-decomposing catalyst, a lignin free radical ion is used as a lignin, and a product having high purity (such as Sigmal, catalog No. 471003, molecular weight: 60,000) which does not contain impurities such as reducing sugar cellulose is used.

1 mL of a solution containing the above lignin: 2.5 mg/mL, 2.5 mM KOH, and porphyrin (protoporphyrin IV manufactured by Sigmal Co., catalog number 258385-1G): 50 μg/mL was placed in Eppendorf, which was made of polypropylene. The tube-transparent cylindrical test tube was irradiated with ultraviolet light for 12 hours using an ENF type ultraviolet lamp manufactured by Spectronix. As a result, it was found that the pH of the lignin reaction solution was lowered from 10.7 to 7.4. Therefore, it is known that free hydrogen ions are derived from lignin.

Further, the use of sunlight instead of ultraviolet light, after repeating the above process, gave the same result.

[Embodiment 14]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 1 mL of a solution containing lignin: 2.5 mg/mL, 0.05 M KOH, and porphyrin: 50 μg/mL obtained in the above Production Example 1 was placed in a light-transmitting cylinder having a degree of Eppendorf tube made of polypropylene. The test tube was irradiated with ultraviolet light for 12 hours using an ENF type ultraviolet lamp manufactured by Spectronix. The resulting sample was analyzed by HPLC using a colloidal filter column. As a control, the solution before ultraviolet irradiation was also analyzed. As a result of the analysis, it was found that 72% of the lignin was decomposed by the peak area of the lignin dissolved in 7 min in the light absorption at 310 nm in the sample after the irradiation. The decomposition product is colloidally filtered to dissolve off the lower molecular weight fraction. Since the absorbance of the decomposition product of lignin in the present invention is low, the absorbance at 310 nm cannot be drawn, and as a non-volatile decomposition product, the eluted sample is freeze-dried, and the same fraction is collected to obtain a precipitate of the decomposition product. .

As a lignin, a product having high purity (such as Aldrich, catalog No. 471046, molecular weight 12,000) and a product insoluble in water (made by Aldrich Co., Ltd., catalog No. 370967), which are impurities containing no reducing sugar or cellulose, are used. After the above process is carried out, the same degree of results are obtained. That is, it is known that, irrespective of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water, when the lignin-base compound solution is further added as a photocatalyst porphyrin, it can be obtained with wood. The same degree of results associated with the decomposition of light.

Further, the use of sunlight instead of ultraviolet light, after repeating the above process, gave the same result.

[Example 15]

As a lignin, a product having high purity (such as Sigmal, catalog No. 471003, molecular weight 60,000) containing no impurities such as reducing sugar or cellulose was used. 0.5 mL of a solution of 50 μg/mL of fecal porphyrin I (manufactured by ALDRICH Co., Ltd.) containing carboxyl 205 mg/mL, 0.05 M KOH, and a synthetic porphyrin, and containing 4 carboxy groups, was allowed to remain in the air space. A 2 mL-capacity was placed in a light-transmissive cylindrical test tube of a polypropylene-made Eppendorf tube, and irradiated with ultraviolet light for 48 hours using an ENF-type ultraviolet lamp manufactured by Spectronix. The reaction liquid thus obtained was treated in the same manner as in Example 2. As a result, it was confirmed that 80% or more of lignin was decomposed.

Further, the space of the air was filled with oxygen, and after irradiation with ultraviolet light as described above, it was confirmed that 80% or more of the lignin was decomposed.

Further, the space of the air was filled with oxygen, and after irradiation with ultraviolet light as described above, it was confirmed that 10% of the lignin was decomposed.

Further, the above porphyrin-containing lignin solution was filled in an amount of no air space in the Eppendorf tube, and almost no decomposition of lignin was observed after irradiation with ultraviolet light as described above.

In addition, as a lignin, a product having high purity (such as Aldrich, catalog No. 471046, molecular weight 12,000) and a product having a slightly lower purity (available from Sigmal Co., Ltd., catalog No.) are used as the lignin. .471038, molecular weight 52,000), and a product insoluble in water (Aldrich, catalog No. 370967), after repeating the above process, the same degree of results were obtained for the decomposition of lignin. That is, regardless of the presence of impurities in the lignin used, the average molecular weight of lignin, or the solubility of lignin in water, lignin can be decomposed to the same extent.

Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained for the decomposition of lignin after repeating the above process.

[Industrial availability]

The catalyst for lignin decomposition of the present invention is not an enzyme such as white decay fungus, and is a catalyst made of non-protein. By using the catalyst, the lignin is mainly decomposed by using light energy, and the lignin which reaches only 20 to 30% of the non-effective resources of the wood can be freely dissolved by an alcohol such as methanol or an organic acid such as formic acid or the like by a simple method. Hydrogen ions can expand the use of lignin, which is difficult to decompose in nature. That is, the alcohol extracted from lignin can be used as a substitute fuel for fossil fuels such as petroleum, and in the field of the fuel industry, organic acids can be used in various industrial fields, and hydrogen ions obtained from lignin, for example, It is used in fuel cells and can be used in the field of the power industry.

Further, since the aromatic hydrocarbon decomposition catalyst of the present invention can be decomposed into aromatic hydrocarbons in which a benzene ring is bonded to an oxygen atom, it is possible to treat harmful industrial waste or to purify contaminated soil or the surface. Therefore, it can be utilized in the field of industrial waste treatment or in the field of purification such as soil.

1 is a structural formula of the product obtained in Production Example 1.

2] The product obtained in Production Example 1 shows the spectrum of the result of the absorbance analysis.

Fig. 3 is a ¹ H NMR spectrum of Sample 1 obtained in Production Example 1.

4 is a ¹³ C NMR spectrum of Sample 1 obtained in Production Example 1.

Fig. 5 is a view showing the HSQC spectrum of Sample 1 obtained in Production Example 1.

Fig. 6 is a COSY spectrum of Sample 1 obtained in Production Example 1.

Fig. 7 is a HMBC spectrum of Sample 1 obtained in Production Example 1.

8] The NOESY spectrum of the sample 1 obtained in Production Example 1.

9 shows the NOESY spectrum of Sample 1 obtained in Production Example 1.

Fig. 10 is a ¹ H NMR spectrum of Sample 2 obtained in Production Example 1.

Fig. 11 is a ¹³ C NMR spectrum of Sample 2 obtained in Production Example 1.

Fig. 12 is an enlarged view showing the NEOSY spectrum of Sample 2 obtained in Production Example 1.

Fig. 13 is a graph showing the relative amount of the sample 2 obtained in Production Example 1.

Fig. 14 shows the results of analysis by ESI-MS.

Fig. 15 shows the results of analysis of the product obtained in Production Example 1 by thermal decomposition GC-MS.

Claims (13)

Use of a porphyrin compound for use in a method for producing a catalyst for lignin decomposition, characterized in that the catalyst for lignin decomposition is a porphyrin containing a catalyst function by light irradiation, and the porphyrin is There is no transition metal atom coordination at the center of the porphyrin ring, and at least one of the carbon atoms of two adjacent pyrrole rings bridged (bridged) is bonded to the hydrogen atom. The use of the porphyrin compound used in the method for producing a catalyst for lignin decomposition according to the first aspect of the invention, wherein the porphyrin is a methyl group and an ethyl ester group or a propionate group in the porphyrin ring. The use of a porphyrin compound for use in a method for producing a catalyst for lignin decomposition according to the first aspect of the invention, wherein the porphyrin contains 4 methyl groups and 4 ethyl ester groups or acetic acid in 4 porphyrin rings. Base or propionate base. The use of the porphyrin compound used in the method for producing a catalyst for lignin decomposition according to the first aspect of the patent application, wherein the porphyrin has two carboxyl groups in the porphyrin ring. The use of the porphyrin compound used in the method for producing a catalyst for lignin decomposition according to the fourth aspect of the patent application, wherein the porphyrin is a carboxyl group having a total of two, four or eight. The use of the porphyrin compound used in the method for producing a catalyst for lignin decomposition according to the fourth aspect of the invention, wherein the porphyrin is at least one selected from the group consisting of uroporphyrin, protoporphyrin, and coproporphyrin. Use of a porphyrin compound for use in a method for producing a catalyst for lignin decomposition, characterized in that the catalyst for lignin decomposition is a porphyrin containing a catalyst function by light irradiation, and the porphyrin is Porphyrin ring The center does not have a transition metal atom coordination, and at least one of the carbon atoms of two adjacent pyrrole rings bridging (bridge) is bonded to a hydrogen atom, and the catalyst is obtained by culturing Escherichia coli in a culture medium. The tetrapyrrole compound having a porphyrin ring structure. The use of the method for producing a catalyst for lignin decomposition according to the seventh aspect of the patent application, wherein the catalyst is a medium containing a tetrapyrrole compound having a porphyrin ring structure obtained by culturing Escherichia coli in a culture medium. Adult. The use of the method for producing a catalyst for lignin decomposition according to the seventh aspect of the patent application, wherein Escherichia coli is an Escherichia coli which is mutated by the gene ypjD (b2611), or the expression of the gene ypjD (b2611) Partial or completely missing state. The use of the method for producing a catalyst for lignin decomposition according to the seventh aspect of the patent application, wherein Escherichia coli is a transposon insertion transgenic mutant of the gene ypjD (b2611). A method for producing a tetrapyrrole compound, characterized in that Escherichia coli which is mutated by the gene ypjD (b2611) is cultured in a medium, and a tetrapyrrole compound having a porphyrin ring structure is obtained from the medium. A method for producing a tetrapyrrole compound according to claim 11 wherein the tetrapyrrole compound is a compound having no transition metal atom coordinated to the center of the porphyrin ring. The method for producing a tetrapyrrole compound according to claim 11 or 12, wherein the tetrapyrrole compound has 4 groups in the porphyrin ring. And a compound of 4 ethyl ester or acetate or propionate.