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
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 Download PDFInfo
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本發明係關於木質素分解用觸媒、醇類及有機酸類之製造方法、木質素分解生成物之製造方法、芳香族烴分解用觸媒、氫離子之游離方法、以及卟啉,特別為由卟啉所成之木質素分解用觸媒、由木質素製造甲醇等醇類及有機酸類之方法、木質素分解生成物之製造方法、由卟啉所成之芳香族烴分解用觸媒、使用由卟啉所成之木質素分解用觸媒由木質素游離氫離子之方法、以及卟啉。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.
進入21世紀後地球上之溫暖化加速度地進行,二氧化碳碳成為控制產業界,甚至成為控制世界經濟之關鍵。只要將封鎖於地中或海底之石化燃料作為能源,無法減少大氣中之二氧化碳碳,難以抑制其增加。於此受到注目者為,由植物製造之生物乙醇等醇類,可望將此作為能源。但,此時過去技術幾乎使用糖類作為原料,故產生人類的食物源與能源之競爭的問題。最近終於不與食料源競爭,例如已進步到將纖維素等作為碳源之醇類製造技術。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.
不能成為木材、或食料之草類主要係由纖維素與木質素所成。藉由作為廢材、片狀建築資材之不適當的接近所謂廢棄物的木材、或草類之纖維素作為碳源時,可抑制二氧化碳碳排出,可貢獻於產業界、經濟界。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.
對於與纖維素的利用如上述進步之比較,與纖維素同樣豐富碳源的木質素之有效利用至今依舊受到極度限制。作為實用段階者,例如作為熱源僅單純燃燒、或防腐劑、或混合於水泥中的結構強化劑等。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.
又,隨著科學技術及產業的發展,產生在自然界中原先非高濃度下存在之化合物的產業廢棄物累積,對於人類成為非常大之負面資產。這些化合物的大部分為,含有於形成苯環之碳原子上結合氧原子之芳香族烴的化合物。例如可舉出戴奧辛類等。有關戴奧辛類,藉由在發生源之垃圾燃燒段階的高溫處理,雖經相當改善,但已經擴散之有害化合物亦不少。因此,將這些有害化合物使用原先存在於自然界的有害化合物分解觸媒,進一步地可利用無限注入於地表的太陽光而分解,其可貢獻自然淨化及人類健康維持者。但至今尚未提出該有用分解觸媒。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.
且,近年來作為能源受到注目者為藉由氫離子之移動得到電動勢之燃料電池。然而,氫氣作為氫離子源時,於現狀氫氣幾乎係由化石燃料所製造。又,藉由水之電分解可得到氫氣,但此時亦必須供給電力。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.
又,電力係由太陽光所得之太陽電池時,半導體裝置之製造為必要,因此若現行能量需要取決於太陽電池時,資源與成本成為莫大的負擔。對於色素增感型太陽電池,奈米尺寸之氧化鈦為必要,且得到一定程度之電動勢的合成色素為高價。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.
過去已知在光照射下將木質素系物質與功能水接觸而分解木質素系物質的分解方法(例如參照專利文獻1)。該過去技術中記載作為功能水之氫氧化鈉,但對於分解物為何物則並無具體記載,對於得到醇類亦無記載。又,對於含有多數個Cl及F的卟啉,已知於木質素之酸化、由鏈烷轉化為醇等反應中具有活性(例如參照專利文獻2)。然而,該過去技術中並無記載有關鹼化合物、光觸媒的使用。 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.
[專利文獻] [Patent Literature]
[專利文獻1]特開2000-144592號公報(申請專利範圍,段落:0030) [Patent Document 1] JP-A-2000-144592 (Patent Application, Paragraph: 0030)
[專利文獻2]特表平2-503086號公報(第4頁左上欄) [Patent Document 2] Japanese Patent Publication No. 2-503086 (the upper left column of page 4)
本發明的課題為,提供將由過去技術上不存在的卟啉所成之木質素分解用觸媒、木質素作為原料,製造甲醇等醇類及有機酸類之方法、製造木質素分解生成物之方法、由卟啉所成之芳香族烴分解用觸媒、將木質素作為原料之游離氫離子的方法、以及卟啉。 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.
本發明的木質素分解用觸媒為係以含有藉由光照射表現觸媒功能之卟啉為特徵。 The catalyst for lignin decomposition of the present invention is characterized by containing a porphyrin exhibiting a catalytic function by light irradiation.
本發明的木質素分解用觸媒又以含有鹼溶液中表現觸媒功能之卟啉為特徵。 The catalyst for lignin decomposition of the present invention is further characterized by a porphyrin containing a catalyst function in an alkali solution.
本發明的木質素分解用觸媒又以含有藉由光照射在鹼溶液中表現觸媒功能的卟啉所成為特徵。 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.
對於上述木質素分解用觸媒係,以卟啉為於(4個)卟啉環含有(4個)甲基及(4個)乙基酯或乙酸基(丙酸基)之四吡咯化合物為特徵。 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.
對於上述木質素分解用觸媒係,以(4個)卟啉為於卟啉環含有4個甲基、4個乙基酯或乙酸基(丙酸基)之四吡咯化合物為特徵。 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.
對於上述木質素分解用觸媒係,以卟啉為分子中具有羧基之卟啉者為特徵。 The above-mentioned catalyst system for lignin decomposition is characterized in that porphyrin is a porphyrin having a carboxyl group in a molecule.
對於上述木質素分解用觸媒係,以卟啉為分子中具有合計2個、4個或8個之羧基的卟啉者為特徵。 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.
對於上述木質素分解用觸媒係,以卟啉為選自尿卟啉、原卟啉、及糞卟啉之至少1種者為特徵。 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.
對於上述木質素分解用觸媒係,以卟啉為具有將大腸桿菌在培養基進行培養,由該培養基所得之卟啉環結構的四吡咯化合物為特徵。 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.
對於上述木質素分解用觸媒係,以該觸媒係含有由將大腸桿菌在培養基進行培養所得之具有卟啉環結構的四吡咯化合物之培養基所成為特徵。 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.
對於上述木質素分解用觸媒係,以大腸桿菌為基因ypjD(b2611)經突變而變的無法表現之大腸桿菌為特徵。 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).
對於上述木質素分解用觸媒係,以大腸桿菌為基因 ypjD(b2611)之轉座子(Transposon)***突變株者為特徵。 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.
上述木質素分解用觸媒係,卟啉為於卟啉環之中心未有過渡金屬原子配位為特徵。 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.
四吡咯化合物之製造方法為,基因ypjD(b2611)藉由突變而變得無法表現之大腸桿菌,或基因ypjD(b2611)之表現機能經部分的或完全欠缺之狀態之大腸桿菌在培養基進行培養,由該培養基所得之四吡咯化合物為特徵。 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.
本發明的醇類及有機酸類之製造方法係,以於木質素加入含有鹼化合物之溶液,由該木質素-鹼化合物溶液分離出醇類及有機酸類者為特徵。 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.
對於上述醇類及有機酸類之製造方法係,以對於木質素-鹼化合物溶液經光照射(較佳為經紫外線或太陽光等照射)後,由該木質素-鹼化合物溶液分離出醇類及有機酸類者為特徵。 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.
對於上述醇類及有機酸類之製造方法係,以對於木質素-鹼化合物溶液,進一步使上述木質素分解用觸媒作用後,由該木質素-鹼化合物溶液分離出醇類及有機酸類者為特徵。 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.
對於上述醇類及有機酸類之製造方法係,以對於木質素-鹼化合物溶液,使上述木質素分解用觸媒作用後,進一步經光照射(較佳為經紫外線或太陽光等照射),由該木質素-鹼化合物溶液分離出醇類及有機酸類者為特徵。 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.
本發明的醇類及有機酸類之製造方法又以對於木質素使上述木質素分解用觸媒作用後,由木質素分離出醇類及有機酸類者為特徵。 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.
本發明的醇類及有機酸類之製造方法又以對於木質素經光照射(較佳為經紫外線或太陽光等照射),由木質素分離出醇類及有機酸類者為特徵。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).
對於上述醇類及有機酸類之製造方法係,以對於木質素使上述木質素分解用觸媒作用後,經光照射(較佳為經紫外線或太陽光等照射),由木質素分離出醇類及有機酸類者為特徵。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.
對於上述醇類及有機酸類之製造方法係,以鹼化合物為KOH及NaOH之至少1種者為特徵。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.
對於上述醇類及有機酸類之製造方法係,以醇類為甲醇,有機酸類為甲酸、乙酸、蘋果酸、琥珀酸及丙酮酸者為特徵。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.
對於上述醇類及有機酸類之製造方法係以醇類的分離藉由蒸餾而進行為特徵。The production method of the above alcohols and organic acids is characterized in that the separation of alcohols is carried out by distillation.
本發明的木質素分解生成物之製造方法係,以依據上述醇類及有機酸類之製造方法,回收分離醇類及有機酸類時所產生的低分子量的含有碳之化合物的木質素分解生成物者為特徵。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.
本發明之形成苯環的碳原子上結合氧原子之芳香族烴分解用觸媒為含有卟啉為特徵。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.
對於上述芳香族烴分解用觸媒係,以卟啉為具有將大腸桿菌在培養基進行培養,由該培養基所得之卟啉環結構的四吡咯化合物為特徵。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.
對於上述芳香族烴分解用觸媒係,以該觸媒由含有具有將大腸桿菌在培養基進行培養所得之卟啉環結構的四吡咯化合物之培養基所成者為特徵。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.
對於上述芳香族烴分解用觸媒係,以大腸桿菌為基因ypjD(b2611)藉由突變而變的無法表現之大腸桿菌為特徵。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).
對於上述芳香族烴分解用觸媒係,以大腸桿菌為基因ypjD(b2611)的轉座子(Transposon)***突變株為特徵。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).
對於上述芳香族烴分解用觸媒係,以卟啉為於卟啉環含有甲基及乙基酯或乙酸基(丙酸基)之四吡咯化合物為特徵。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.
對於上述芳香族烴分解用觸媒係,以卟啉為於卟啉環含有4個甲基、4個乙基酯或乙酸基(丙酸基)之四吡咯化合物為特徵。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.
對於上述芳香族烴分解用觸媒係,以卟啉為選自尿卟啉、原卟啉、糞卟啉、及初紫質(etioporphyrin)之至少1種為特徵。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.
對於上述芳香族烴分解用觸媒係,以卟啉為分子中具有羧基之卟啉者為特徵。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.
對於上述芳香族烴分解用觸媒係,以卟啉為分子中具有合計2個、4個或8個之羧基的卟啉者為特徵。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.
對於上述芳香族烴分解用觸媒係,以芳香族烴為戴奧辛類為特徵。The above-mentioned aromatic hydrocarbon decomposition catalyst system is characterized in that the aromatic hydrocarbon is a dioxin.
本發明的氫離子之游離方法係,以對於木質素或木質素-鹼化合物溶液,使上述木質素分解用觸媒作用,對於該溶液進行光照射(較佳為經紫外線或太陽光等照射),使氫離子游離者為特徵。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.
本發明的卟啉係以具有將木質素轉換為醇類及有機酸類之觸媒功能者為特徵。The porphyrin system of the present invention is characterized by having a catalytic function of converting lignin into an alcohol and an organic acid.
本發明的卟啉又係以具有分解含有於形成苯環之碳上結合氧原子之芳香族烴的化合物之觸媒功能者為特徵。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.
本發明的卟啉為藉由將基因ypjD(b2611)藉由突變無法表現的大腸桿菌進行培養而得者。The porphyrin of the present invention is obtained by culturing the gene ypjD (b2611) by Escherichia coli which cannot be expressed by mutation.
本發明的木質素分解用觸媒進一步特徵為,含有將基因ypjD(b2611)藉由突變無法表現的大腸桿菌進行培養所得之卟啉者。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.
本發明為,卟啉例如具有使用大腸桿菌以生物學所製造之卟啉結構之四吡咯化合物或合成卟啉可達成作為有效木質素分解用觸媒及芳香族烴分解用觸媒之效果。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.
本發明對於木質素藉由單獨或組合鹼化合物、光照射、上述木質素分解用觸媒使其作用,由木質素可製造醇類及有機酸類之同時,可得到低分子量之分解生成物,且可達到游離氫離子之效果。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.
以下對於本發明之實施形態做詳細說明。Hereinafter, embodiments of the present invention will be described in detail.
所謂有關於形成本發明之木質素分解用觸媒及苯環的碳原子上結合氧原子之芳香族烴分解用觸媒的一實施形態,這些觸媒含有藉由光照射及/或在鹼溶液中表現觸媒功能之卟啉所成,該卟啉係以具有將大腸桿菌在培養基進行培養,分泌於培養基中之卟啉環結構的四吡咯化合物或合成卟啉為佳。例如這些觸媒亦可為由含有具有將大腸桿菌在培養基進行培養所得之卟啉環結構的四吡咯化合物之培養基所成者,亦可為由進行培養所得之細胞經回收者、或由進行培養所得之細胞而得之萃取物亦可。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.
上述大腸桿菌以如基因ypjD(b2611)經突變而變的無法表現之大腸桿菌的基因表現經變化之大腸桿菌為佳,又基因ypjD(b2611)的轉座子(Transposon)***突變株者為更佳。上述卟啉為,於卟啉環含有甲基(例如4個)、乙酯或乙酸基(丙酸基)(例如4個)之四吡咯化合物為更佳。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.
有關本發明來自大腸桿菌之木質素分解用觸媒及芳香族烴分解用觸媒,例如可由下述製造。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.
作為製造上述觸媒之培養基,僅為可培養大腸桿菌之培養基即可,並無特別限制而可使用。將大腸桿菌在貧營養培養基或富營養培養基進行培養,由該培養基分離並回收四吡咯化合物,可製造出具有構成木質素分解用觸媒及芳香族烴分解用觸媒之卟啉環結構的四吡咯化合物。在培養基中將大腸桿菌進行培養、增殖的過程中,使大腸桿菌產生四吡咯化合物,藉由回收於培養基中分泌之四吡咯化合物,製造出四吡咯化合物。欲防止培養基中所具有之天然物等成分成為分離四吡咯化合物時的障礙,使用貧營養培養基為佳,但對於該培養基並無特別限定。作為貧營養培養基,可使用含有葡萄糖或乳糖者為佳,但未限定於此等。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.
製造木質素分解用觸媒及芳香族烴分解用觸媒時所使用的大腸桿菌以基因ypjD(b2611)經突變而變的無法表現者為佳。例如可舉出來自K12株及BL21株之大腸桿菌等。例如使用來自K12株之基因ypjD(b2611)經突變無法表現之大腸桿菌為佳。作為基因ypjD(b2611)經突變而變的無法表現之大腸桿菌株,例如有基因ypjD(b2611)之轉座子(Transposon)***突變株。該突變株為基因ypjD(b2611)之表現經部分的或完全欠缺之狀態。且,K12株例如可由National BioResource得到,BL21株例如可由Takarabio得到。又,作為基因ypjD(b2611)之轉座子(Transposon)***突變株,例如可由National BioResource得到之JD23504等。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.
對於本實施之形態,首先將大腸桿菌在貧營養培養基中進行培養。此時,將大腸桿菌在貧營養培養基以外的適當培養基,例如LB培養基等培養基中進行前培養,將所得之前培養物接種於貧營養培養基進行本培養為佳。又,作為培養基,可使用貧營養培養基以外之富營養培養基或合成培養液等。例如亦可使用於脫離子水中加入KH2PO4、K2HPO4、(NH4)2SO4、檸檬酸二水合物、葡萄糖、及MgSO4等所得之水溶液的合成培養液。僅為可增加大腸桿菌細胞之培養基即可,可為任一種並無特別限定。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.
大腸桿菌的培養條件為,對於大腸桿菌之一般條件即可。此為例如將大腸桿菌進行前培養後改變培養基在貧營養培養基中進行本培養時,對於任合培養條件皆相同。例如使用LB培養基,在15℃~40℃之溫度進行6小時~24小時之前培養後,將所得之細胞懸濁液於貧營養培養基中,進行20℃~40℃之溫度下12小時~96小時之本培養。藉此,在培養基中可增殖細胞,得到目的之帶有四吡咯化合物之特有色調的培養物(產物)。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.
其次,由上述培養物如以下分離出作為目的之四吡咯化合物。Next, the intended tetrapyrrole compound was isolated from the above culture as follows.
具體而言,將離心分離培養物所得之澄清液進行過濾後,例如使用離子交換樹脂管柱或逆相管柱等,自濾液吸附,分離四吡咯化合物。例如將培養物經離心分離器沈澱細胞,得到含有培養物(產物)之澄清液。其次,將該澄清液以所定孔徑(例如0.22μm)之濾器進行過濾後,使用上述管柱吸附於離子交換樹脂。其次,例如使用20%乙腈-0.1%三氟乙酸溶液等由離子交換樹脂溶離產物後,進行冷凍乾燥。且此時之溶離為,可使用於有機溶劑含有酸或鹼之溶液者。所謂本實施之形態,可得到1種類或2種類以上之四吡咯化合物,例如可由500mL之細胞懸濁液得到數mg~數十mg之四吡咯化合物。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.
經藉由上述操作所分離之產物藉由NMR(Nuclear Magnetic Resonance;核磁共振)等進行分析時,確認含有四吡咯化合物。又,將該產物經吸光光度分析時,得知色素為特有波長區域具有吸收的化合物。大多情況為,與葉綠素、原血紅素或酞青素類似之顯示二峰性波峰的色素化合物。如此色素化合物作為藉由光使電子激起的光觸媒或電子傳達體使用時為有用。又,於水溶液中、或介著細胞膜與氧化還原反應相關,故亦可考慮於電池亦可發揮其功能。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.
如上述,使用大腸桿菌,可製造卟吩、卟啉等卟啉類之四吡咯化合物,故如藉由化學合成法之情況,配合目的之化合物種類之製造裝置或觸媒等為必要,又無須使用溶劑,對於環境產生壞影響之顧慮亦較少。又,培養大腸桿菌時,培養基中無須加入5-胺酮戊酸等四吡咯化合物的前驅物(特開平5-244937號公報),且四吡咯化合物為回收分泌於培養基中者即可,無須由菌體採取(特開平5-91866號公報)。即,對於大腸桿菌之培養或四吡咯化合物之回收無須特定化合物或裝置,故可簡便地製造四吡咯化合物。如此所得之四吡咯化合物可利用於醫療、食品及電子學等種種產業領域中之用途。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.
對於上述,由培養物分離四吡咯化合物,作為木質素分解用觸媒及芳香族烴分解用觸媒。使用由培養物回收之化合物為佳,但培養物或培養所得之細胞因含有四吡咯化合物,可將該培養物自體或細胞自體作為木質素分解用觸媒及芳香族烴分解用觸媒使用。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.
作為本發明中可使用的木質素分解用觸媒及芳香族烴分解用觸媒,作為上述其他合成卟啉,例如可舉出選自原卟啉、尿卟啉、糞卟啉、及初紫質等至少1種。但,初紫質(aetioporphyrin)不能分解木質素。作為這些卟啉,在以下實施例,使用分子內含有2個羧基之原卟啉IX(ALDRICH公司製),分子內含有8個羧基之尿卟啉I(SIGMA公司製),分子內含有4個羧基之糞卟啉I(ALDRICH公司製)、及分子內未含有羧基之初紫質(ALDRICH公司製)。該卟啉亦與培養上述大腸桿菌所得之卟啉同樣地,於卟啉環之中心未有過渡金屬原子配位。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.
其次,對於有關本發明之醇類及有機酸類的製造方法之實施形態做說明。本發明中的醇類及有機酸類為,對於木質素以單獨或組合鹼化合物、光照射、上述木質素分解用觸媒方式使其作用而可製造。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.
即,甲醇等醇類為,例如(1)加入於木質素含有選自KOH及NaOH等之至少1種鹼化合物的溶液,由該木質素-鹼化合物溶液例如藉由蒸餾等分離醇類、(2)加入於木質素含有上述鹼化合物之溶液,對於該木質素-鹼化合物溶液將紫外線(作為紫外線,例如可使用來自Spectronix公司製之ENF型(260c/j、或280c/j等)或UVP公司製之UVL-56Hand Held等紫外線燈的紫外線)或太陽光等具有廣波長區域之光以所定時間進行照射後,由照射後之木質素-鹼化合物溶液例如藉由蒸餾等,可有效率地分離醇類、(3)加入於木質素含有上述鹼化合物之溶液,對於該木質素-鹼化合物溶液將上述木質素分解用觸媒在所定溫度下,使其進行所定時間之作用後,由溶液例如藉由蒸餾瞪可有效率地分離出醇類、(4)加入於木質素含有上述鹼化合物之溶液,對於該木質素-鹼化合物溶液將上述木質素分解用觸媒在所定溫度下進行所定時間之作用後,對於該溶液以紫外線或太陽光等具有廣波長區域的光進行所定小時照射,其次由照射後之木質素-鹼化合物溶液例如藉由蒸餾等,可有效率地分離醇類、(5)對於木質素將上述木質素分解用觸媒在所定溫度下進行所定時間之作用後,由該溶液例如藉由蒸餾等,可有效率地分離醇類、(6)對於木質素以紫外線或太陽光等具有廣波長區域之光進行所定時間的照射,其次由該溶液例如藉由蒸餾等,可有效率地分離醇類、以及而(7)對於木質素將上述木質素分解用觸媒在所定溫度下進行所定時間之作用後,對於該溶液以紫外線或太陽光等具有廣波長區域的光進行所定時間的照射,其次由照射後之木質素-鹼化合物溶液例如藉由蒸餾等,可有效率地分離醇類而製造。進行上述光照射時,將含有木質素之反應液與含有空氣或氧、氧及/或氮的氣體等接觸而在環境氣體中進行時,可有效率地發揮卟啉之觸媒功能故較佳。例如以木質素濃度2.5mg/mL、卟啉50μg/mL(質量比50:1)時,每木質素1mg可使用0.2~0.5mL程度之氧。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.
又,甲酸、乙酸、蘋果酸、琥珀酸及丙酮酸等有機酸類與上述醇類之情況相同,可由木質素分離。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.
作為本發明所使用的木質素,並無特別限制,例如可使用未含有還原糖或纖維素等雜質的純度高之製品(例如SIGMA公司製的目錄No.471003,分子量60,000;ALDRICH公司製,目錄No.471046,分子量12,000)、含有還原糖而純度稍低的製品(例如SIGMA公司製之目錄No.471038,分子量52,000)、及不溶於水的製品(例如ALDRICH公司製之目錄No.370967)等。本發明為,由所有這些木質素可分離出幾乎同程度的甲醇等醇類以及甲酸、乙酸、蘋果酸、琥珀酸及丙酮酸等有機酸類。即,與雜質的存在、或平均分子量、對水之溶解性無關,僅為木質素可藉由本發明分離出醇類及有機酸類而製造。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.
作為鹼化合物之溶液,並無特別限制,例如可使用0.0025M~0.05M程度之KOH及/或NaOH等為佳。於醇及有機酸的分離效率上有高低者,鹼化合物之溶液於該濃度範圍內時並無限定。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.
於形成本發明之苯環的碳原子上結合氧原子之芳香族烴分解用觸媒的一實施形態為,該觸媒為具有與含有上述卟啉所成之木質素分解用觸媒相同構成者,詳細說明則省略。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.
作為使上述觸媒起作用之芳香族烴,例如可舉出戴奧辛類或戴奧辛類似化合物等。該戴奧辛類中例如含有多氯二氧化二苯(PCDD)、多氯二苯並呋喃(PCDF)等,戴奧辛類似化合物中例如含有多氯聯苯(戴奧辛樣PCB)等。所謂本發明之芳香族烴分解用觸媒為這些戴奧辛類經分解所得到之無毒化者。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.
且,所謂本發明之其他實施形態,由木質素游離氫離子之方法中,藉由添加於木質素-鹼化合物溶液含有來自上述大腸桿菌的吡咯化合物所成之卟啉或合成卟啉而成的木質素分解用觸媒,游離氫離子之同時,可進行木質素之光分解。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.
且,所謂本發明之其他實施形態,依據上述醇類及有機酸類的製造方法,可回收分離醇類及有機酸類時所產生的木質素分解生成物之含有低分子量碳的化合物。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.
本發明並未限定於上述實施形態,在不脫離本發明主旨的範圍內,可實施種種變化形式。以下對於本發明之製造例及實施例做詳細說明。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.
如上述,於木質素藉由作為木質素分解用觸媒添加卟啉(吡咯化合物),可製造出醇類及有機酸類,進行木質素之光分解而可得到分解生成物之同時,可游離氫離子。又,具有卟啉結構之四吡咯化合物或合成卟啉作為對於戴奧辛等芳香族烴之分解用觸媒時有效。此時作為卟啉,例如可使用以使用大腸桿菌的生物學方式製造者。本製造例中,對於使用大腸桿菌之吡咯化合物的製造例做說明。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.
首先,將來自大腸桿菌之基因ypjD(b2611)的轉座子(Transposon)***突變株(National BioResourceJD23504)的細胞,於2mL的LB培養基(Bacto-tryptone:1%,Bacto-yeast extract:0.5%,NaCl:0.5%)中,在37℃下進行12小時的前培養。將所得之細胞懸濁液1mL,加入於脫離子水1L中KH2PO4為9g、K2HPO4為21g、(NH4)2SO4為2g、檸檬酸二水合物為1g、葡萄糖為3.6g、及MgSO4為200mg所得之水溶液500mL中,在37℃進行24小時主要培養。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.
所得之培養液的顏色為,開始主要培養的時點為無色,但經過24小時後成為粉紅色。將該培養液經離心分離器使細胞沈澱,將所得之澄清液以孔徑0.22μm之濾器進行過濾。其次,將濾液通過陰離子交換樹脂填充管柱後,將吸著於樹脂的培養物以20%乙腈-0.1%三氟乙酸溶液進行溶離,再將溶離物進行冷凍乾燥。藉此,得到帶有粉紅色之產物。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.
對於該產物,進行下述所記載的各種機器分析時,確認為具有圖1所示結構的四吡咯化合物。圖1中的記號A~G為對應圖4所示13C NMR光譜的記號,對於其他圖面亦相同。又,將產物藉由NMR進行分析時,確認含有四吡咯化合物。且,藉由ICP(Inductively Coupled Plasma)質量分析檢測出鉀(K),作為形成K與離子結合或錯體之化合物而回收。且進行吸光光度分析時,如圖2所示,確認含有索瑞特帶(Soret band)之卟啉特有二峰性波峰。圖2中,波長395nm與波長549nm各有波峰。由此確認出產物為自由電子可移動之有機色素。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.
將上述所得之四吡咯化合物溶解於相異溶劑(試料1及試料2)中,進行二次元NMR測定(COSY、NOESY、HSQC、HMBC),解析該光譜,並進行結構解析。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.
有關試料1,將試料溶解於CD3OD,以以下條件下實施NMR測定。In the sample 1, the sample was dissolved in CD 3 OD, and NMR measurement was carried out under the following conditions.
‧裝置:INOVA500型(varian公司製)‧Device: INOVA500 (variance company)
‧共振頻率:499.8MHz(1H)‧Resonance frequency: 499.8MHz ( 1 H)
‧基準:3.31ppm(CD2HOD、1H NMR)49.421ppm(CD3OD、13C NMR)‧Base: 3.31 ppm (CD 2 HOD, 1 H NMR) 49.421 ppm (CD 3 OD, 13 C NMR)
‧累積次數:1H NMR(16次)、13C NMR(53428)、COSY(16次)、NOESY(8次)、HSQC(32次)、HMBC(128次)‧ Cumulative number: 1 H NMR (16 times), 13 C NMR (53428), COSY (16 times), NOESY (8 times), HSQC (32 times), HMBC (128 times)
‧其他:NOESY之混合時間設定為400msec。‧Other: The mixing time of NOESY is set to 400msec.
又,有關試料2,將試料溶解於CD3CN:D2O:CD3COOD=90:10:0.1之溶劑,在以下條件下實施NMR測定。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.
‧裝置:INOVA600型(varian公司製)‧Device: INOVA600 type (varian company)
‧共振頻率:599.8MHz(1H)‧Resonance frequency: 599.8MHz ( 1 H)
‧基準:1.92ppm(CD2HCN、1H NMR)1.28ppm(CD3CN、13C NMR)‧Base: 1.92 ppm (CD 2 HCN, 1 H NMR) 1.28 ppm (CD 3 CN, 13 C NMR)
‧累積次數:1H NMR(64次)、13C NMR(50000)、COSY(16次)、NOESY(16次)、HSQC(32次)、HMBC(128次)‧ Cumulative times: 1 H NMR (64 times), 13 C NMR (50000), COSY (16 times), NOESY (16 times), HSQC (32 times), HMBC (128 times)
‧其他:NOESY的混合時間設定為400msec。‧Other: The mixing time of NOESY is set to 400msec.
圖3表示1H NMR光譜(溶劑:CD3OD)。其結果於10.0~10.5ppm附近(d)、4.3ppm附近(f)、3.6ppm附近(g)、及3.2ppm附近(e)觀測到推測為來自目的成分之訊號。各訊號強度比約為1:2:3:2。該d~g於其他圖面上皆相同。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.
圖4表示13C NMR光譜。推定為自B、C的訊號之芳香族。Figure 4 shows the 13 C NMR spectrum. It is presumed to be the aromatic of the signal from B and C.
其中,1H NMR之d訊號由化學位移值來看,一般為無法見到的特徵性訊號,且若考慮為芳香族化合物時,則可能為卟啉骨格。如以下所述,將二次元NMR光譜(圖5~9)作為卟啉進行解析時可無矛盾下而解析。又,與圖1之推定結構類似的化合物於J. org. Chem. Vol. 164,No. 21,1999(7973-7982)已被報告,1H NMR的化學位移值顯示相當一致,故推定為卟啉結構較為適當。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.
以下對於二次元NMR之解析進行說明。The analysis of the binary NMR will be described below.
圖5表示HSQC光譜。HSQC光譜為檢測J1 CH的測定法。將解析結果記載於圖中。對於質子訊號,在直接鍵結的碳號碼之大文字上記載號碼。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.
圖6表示COSY光譜。COSY光譜為檢測1H-1H間之自旋偶合的測定法。解析結果得知f與e為自旋偶合,由訊號強度比及F與E之化學位移值得知為-CH2-CH2-X之可能性高。其中,X表示結構不明之未確定成分。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.
圖7表示HMBC光譜。HMBC光譜藉由檢測Jn CH(n=2~4程度)之測定法,得到異種核遠隔結合相關光譜。該光譜經解析之結果得到(e,A)、(e,B)、(e,F)、(g,B)、(g,C)、(f,A)、(f,B)、(f,C)、(f,E)等相關。這些相關未與圖1所示推定結構相矛盾。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.
圖8及圖9表示NOESY光譜。NOESY光譜為檢測磁化交換的測定法,由藉由交差緩和之磁化移動可得到核自旋間之距離相關情報。光譜經解析之結果為於(g,e)、(f,g)、(f,e)、(d,f)、(d,e)、(d,g)觀測到NOE相關。這些NOE相關為支持圖1所示推定結構者。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.
圖10表示1H NMR光譜,圖11表示13C NMR光譜(溶劑:CD3CN:D2O:CD3COOD=90:10:0.1)。圖10中以四角框圍的訊號表示雜質。與上述試料1之光譜比較結果,推測主成分之結構相同。此亦可由COSY、NOESY、HSQC、及HMBC之各光譜的解析結果得到支持。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.
圖12表示擴大試料2之NOESY光譜。因觀測到d質子分離為4種,故自低磁場(數字較小者)而賦予d1至d4之號碼。歸納NOE相關如以下。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及d4同時與甲基及-CH2-CH2-X為NOE相關。‧ d1 and d4 are simultaneously associated with methyl and -CH 2 -CH 2 -X for NOE.
‧ d2僅與-CH2-CH2-X為NOE相關。‧ d2 is only associated with -CH 2 -CH 2 -X for NOE.
‧ d3僅與甲基為NOE相關。‧ d3 is only related to methylation as NOE.
此與未明確觀測到甲基彼此或-CH2-CH2-X彼此之NOE相關,而得到圖1之側鏈配置。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.
13C訊號及1H訊號的號碼為幾乎相同區域所觀測之訊號賦予一號碼。此理由為,卟啉之骨格為重複結構,故難以詳細地歸類。有關重複個數,因觀測到4條甲基(g),故推測有4個。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.
如上述,由試料1及2之分析結果得知,可得到圖1之結構。但,側鏈的甲基及-CH2-CH2-X之各合計為4個即可,附加部位如圖1所示8處中任一處皆未與數據矛盾,未限定於圖1所示之附加位置。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.
其次,對於相當於-CH2-CH2-X中之X部分做檢討。Next, a review is made for the X portion corresponding to -CH 2 -CH 2 -X.
對於上述所得之產物進行以下分析。The following analysis was carried out for the product obtained above.
與除去TFA等,將試料在加熱爐內,加熱280℃×10min後,在600℃進行熱分解,將分解物以下述所記載之氣體色層/質量分析裝置(以下稱為「GC/MS」)進行分離分析。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.
裝置名:Agilent Technologies製HP5973型 附有質量選擇型檢測器Device name: HP5973 model manufactured by Agilent Technologies with quality selective detector
HP6890型 氣體色層HP6890 gas color layer
FRONTIER LAB製PY-2020iD加熱爐式熱分解裝置FRONTIER LAB PY-2020iD heating furnace type thermal decomposition device
測定試料:將試料2mg中添加AcCN 1mL的溶液注入於試料杯中0.5mL,再以氮氣沖洗將AcCN除去。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.
欲調查溶液中成分之分子量,藉由下述記載的電噴灑-質量分析裝置(以下稱為「ESI-MS」)進行分析。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.
裝置名:Applied Biosystems製QstarDevice name: Qstar made by Applied Biosystems
導入溶劑:AcCN/0.05%甲酸水溶液(50/50)Introduced solvent: AcCN/0.05% formic acid in water (50/50)
導入方法:使用30μL之環導線將測定溶液直接導入。Introduction method: The measurement solution was directly introduced using a 30 μL loop wire.
測定模式:Positive modeMeasurement mode: Positive mode
測定溶液:將於試料2mg加入AcCN 1mL的溶液於小玻璃瓶少量採取後,以導入溶劑稀釋至約100ppm。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.
以上述GC/MS所進行的分析結果,由試料檢測出二氧化碳碳。又,因以上述ESI-MS所進行的分析結果(圖14),檢測出分子量59之片段離子(波峰為4條),確認該片段為含有乙基酯或乙酸基(丙酸基)。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).
且,將上述產物經熱分解GC-MS分析結果,如圖15所示,主要成分為吡咯化合物。此可由將各成分的質量光譜與數據基值做對照而確認,可確認分子中含有吡咯環之結構。又,對於詳細分子量,如圖13所示,檢測出附加氫離子的離子,得到分子量655.2760-1.0078=654.2682。由以上可確認於側鏈含有4個乙基酯或乙酸基(丙酸基),4個甲基之分子量654,分子式C36H38O8N4之卟啉化合物,又確認於卟啉環之中心未有過渡金屬。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.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:2.5mg/mL、0.05M KOH、及依據上述製造例1所得之分子中具有4個羧基之卟啉:50μg/mL的溶液1mL,放於持有聚丙烯製Eppendorf tube程度的光透過性之圓筒形試管中,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。其後,以80℃進行60分鐘加熱,經揮發之氣體使用島津GC/MS QP-2010,管柱DB-WAX進行分析,測定到170μg/mL之甲醇。此時,得到甲酸為140μg/mL,蘋果酸為25μg/mL,乙酸為19μg/mL,琥珀酸為5.4μg/mL,丙酮酸為6.2μg/mL。此時,以乾燥木質素重量基準下,可得到相當於6.8重量%之甲醇,相當於5.6重量%之甲酸。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000),含有還原糖而純度稍低之製品(Sigmal公司製,目錄No.471038,分子量52,000)、及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到作為甲醇量及上述有機酸量之同程度的結果。即,與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,得知自木質素可分離製造醇類及有機酸類。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.
且,取代紫外光使用太陽光時,重複上述製程後,得到作為甲醇量及有機酸量之同樣結果。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.
將如上述經紫外光照射所得之反應液樣品於HPLC(高速液體色層法),使用膠體過濾管柱進行分析。作為對照,對於紫外線照射前之溶液亦進行分析。分析結果,照射前時木質素於310nm之吸光中,檢測出在7-9min經溶離之波峰,但如上述,於木質素添加卟啉,以紫外光照射轉換為甲醇或有機酸類時,310nm之吸光中,在7-9min經溶離的木質素之波峰面積縮小為20%。即,得知於木質素添加卟啉,以紫外光進行照射時,80%之木質素會被分解。該分解生成物藉由膠體過濾,溶離出較低分子量的分餾物。因此,以經分解之木質素基準下,可得到相當於約8.5重量%之甲醇、約相當於7重量%之甲酸。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.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:2.5mg/mL、0.05M KOH、及作為合成卟啉之分子內含有2個羧基之原卟啉IX(ALDRICH公司製)、分子內含有8個羧基之尿卟啉I(SIGMA公司製)、分子內含有4個羧基之糞卟啉I(ALDRICH公司製)之各50μg/mL的各溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。將所得之反應液在80℃進行60分鐘加熱,將揮發之氣體使用島津GC/MS QP-2010,管柱DB-WAX進行分析。重複上述操作數次後,依據原卟啉IX、尿卟啉I、及糞卟啉I之各情況,以乾燥木質素重量基準下,得到幾乎與實施例1之情況同等量之甲醇及甲酸。即,以乾燥木質素重量基準下,約6~9重量%轉換為甲醇,約2~4重量%轉換為甲酸。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.
將對於如上述所得之各反應液的樣品以HPLC(高速液體色層法)使用膠體過濾管柱進行分析。作為對照,對於紫外線照射前之溶液亦進行分析。分析結果,照射前的情況為,木質素於310nm的吸光中,檢測出以7-9min經溶離的波峰,但於木質素添加上述各卟啉,照射紫外光轉換為醇及有機酸類(甲酸等)的反應液中,原卟啉IX的情況為約縮小至60%,尿卟啉I的情況為約縮小至15%,糞卟啉I的情況為約縮小為20%。即,得知於木質素添加卟啉,以紫外光照射時,原卟啉IX的情況為分解約40%,尿卟啉I的情況為分解約85%,糞卟啉I的情況為分解約80%的木質素。該分解生成物藉由膠體過濾可溶離出較低分子量的分餾物。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、含有還原糖而純度稍低之製品(Sigmal公司製,目錄No.471038,分子量52,000)、及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,使用合成卟啉之情況,亦對木質素之分解得到相同程度之結果。即,與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,可分解相同程度之木質素。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.
且,取代紫外光使用太陽光時,重複上述製程後,可得到對於木質素分解之同樣結果。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.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:1.8mg/mL、0.05M KOH、及Sigmal公司製之原卟啉IV(目錄No.258385-1G):50μg/mL的溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。其後,以80℃進行60分鐘加熱,將揮發之氣體使用島津GC/MS QP-2010、管柱DB-WAX進行分析時,測定出54μg/mL之甲醇。即,以乾燥木質素重量基準下,可得到3.0重量%之甲醇。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、含有還原糖而純度稍低之製品(Sigmal公司製,目錄No.471038,分子量52,000)、及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到作為甲醇量之相同程度的結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,由木質素分離製造醇類。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.
且,取代紫外光使用太陽光時,重複上述製程後,得到作為甲醇量之同樣結果。Further, when sunlight was used instead of ultraviolet light, the same results as the amount of methanol were obtained after repeating the above process.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:7.5mg/mL及0.05M KOH之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,在80℃進行60分鐘加熱,將揮發之氣體使用島津GC/MS QP-2010,管柱DB-WAX進行分析後,測定出93μg/mL之甲醇。即,以乾燥木質素重量基準下,可得到1.24重量%之甲醇。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、以及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到作為甲醇量相同程度之結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,可由木質素製造相同程度之甲醇。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.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:7.5mg/mL及0.05M KOH之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射24小時。其後,以80℃進行60分鐘加熱,將揮發氣體使用島津GC/MS QP-2010,管柱DB-WAX進行分析時,測定出150μg/mL之甲醇。即,可得到以乾燥木質素重量基準下2重量%之甲醇。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、以及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到作為甲醇量相同程度之結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,由木質素製造相同程度之甲醇。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.
且,取代紫外光使用太陽光時,重複上述製程後,得到作為甲醇量之同樣結果。Further, when sunlight was used instead of ultraviolet light, the same results as the amount of methanol were obtained after repeating the above process.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:2.5mg/mL、0.05M KOH、及依據上述製造例1所得之卟啉:25μg/mL的溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。其後,以80℃進行60分鐘加熱,將揮發之氣體使用島津GC/MS QP-2010,管柱DB-WAX進行分析,測定出160μg/mL之甲醇。即,以乾燥木質素重量基準下,可得到6.4重量%之甲醇。得知藉由於木質素-鹼化合物溶液進一步添加作為光觸媒之卟啉,可飛躍式地促進由木質素之甲醇的製造量。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、以及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到作為甲醇量相同程度之結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,可由木質素製造相同程度之甲醇。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.
且,取代紫外光使用太陽光時,重複上述製程後,作為甲醇量得到同樣結果。Further, when sunlight was used instead of ultraviolet light, the same result was obtained as the amount of methanol after repeating the above process.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:2.5mg/mL之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈照射紫外光24小時。其後,以80℃進行60分鐘加熱,將揮發之氣體使用島津GC/MS QP-2010,管柱DB-WAX進行分析時,測定出21μg/mL之甲醇。即,以乾燥木質素重量基準下可得到0.84重量%之甲醇。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、以及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到作為甲醇量相同程度之結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、與木質素對水之溶解性無關,由木質素可得到相同程度之甲醇。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.
且,取代紫外光使用太陽光時,重複上述製程後,對於甲醇量得到同樣結果。Further, when sunlight is used instead of ultraviolet light, the same result is obtained for the amount of methanol after repeating the above process.
本實施例中,對於苯環上結合氧原子之芳香族烴,使用本發明的芳香族烴分解用觸媒之分解處理方法做說明。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.
使用含有於苯環結合氧原子之芳香族烴的化合物之Remazol Brilliant Blue(RBBR;SIGMA公司製),進行其光分解。該RBBR為作為分解戴奧辛類之指標所使用的類似試藥。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.
將含有RBBR:250μg/mL、0.05M KOH、及依據上述製造例1所得之卟啉、原卟啉IX(ALDRICH公司製)、尿卟啉I(SIGMA公司製)、及糞卟啉I(ALDRICH公司製)之各25μg/mL的溶液(作為溶劑使用30%乙腈、0.1%三氟乙酸、60%甲醇)1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製的ENF型紫外光燈以紫外光照射2小時。其結果,於使用任何卟啉及溶劑之情況下,得知RBBR的藍色調消失,RBBR被分解。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.
且,取代紫外光使用太陽光時,重複上述製程後,同樣可確認RBBR之分解。Further, when sunlight is used instead of ultraviolet light, the decomposition of RBBR can be confirmed by repeating the above process.
本實施例中,使用含有於苯環上結合氧原子之芳香族烴的化合物之氰基二甲苯(Takara公司製)及溴酚藍(Takara公司製),進行該光分解。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.
將含有氰基二甲苯:5mg/mL及溴酚藍:5mg/mL、0.05M KOH、以及依據上述製造例1所得之卟啉、原卟啉IX(ALDRICH公司製)、尿卟啉I(SIGMA公司製)、糞卟啉I(ALDRICH公司製)及初紫質(ALDRICH公司製)之各40μg/mL的溶液(作為溶劑使用30%乙腈、0.1%三氟乙酸)1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射2小時。其結果,於使用任一卟啉及溶劑時,皆為氰基二甲苯及溴酚藍的藍色以及黃色色調消失,得知氰基二甲苯及溴酚藍被分解。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.
且,取代紫外光使用太陽光時,重複上述製程後,同樣可確認氰基二甲苯及溴酚藍之分解。Further, when sunlight is used instead of ultraviolet light, the decomposition of cyanoxylene and bromophenol blue can be confirmed by repeating the above process.
本實施例中,使用本發明之木質素分解用觸媒,由木質素游離氫離子之方法中,作為木質素使用未含有還原糖纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。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).
將含有該木質素2.5mg/mL、及依據上述製造例1所得之卟啉:50μg/mL之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。其結果,木質素反應液之pH自9.2降至6.4。因此得知由木質素可游離氫離子。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、含有還原糖而純度稍低之製品(Sigmal公司製,目錄No.471038,分子量52,000)、及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,同樣地游離氫離子。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,由木質素可游離相同程度之氫離子。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.
且,取代紫外光使用太陽光時,重複上述製程後,得到同樣結果。Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained after repeating the above process.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:2.5mg/mL、2.5mM KOH、及依據上述製造例1所得之卟啉:50μg/mL之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。將該樣品以HPLC(高速液體色層法),使用膠體過濾管柱進行分析。作為對照,對於紫外線照射前之溶液亦進行分析。分析結果為,照射前時,於波長310nm之吸光中,檢測出在7-9min所溶離之波峰,但於木質素添加卟啉,以紫外光照射時,於310nm之吸光中,在7-9min所溶離之木質素的波峰面積縮小至78%。即,得知於木質素添加卟啉,以紫外光照射時,可分解22%之質素。又,木質素反應液的pH自10.7降至6.2。因此得知由木質素游離氫離子。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、含有還原糖而純度稍低之製品(Sigmal公司製,目錄No.471038,分子量52,000)、及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到同樣結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,將木質素以同程度進行分解之同時,可將氫離子以同程度下進行游離。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.
且,取代紫外光使用太陽光時,重複上述製程後,得到同樣結果。Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained after repeating the above process.
使用本發明之木質素分解用觸媒,自木質素游離氫離子之方法中,作為木質素使用含有還原糖而純度稍低之製品(Sigmal公司製,目錄No.471038,分子量52,000)。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.
將含有上述木質素5mg/mL及依據上述製造例1所得之卟啉化合物50μg/mL之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。其結果木質素反應液的pH自6.2降至4.8。因此,得知由木質素游離氫離子。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000、及Aldrich公司製,目錄No.471046,分子量12,000)、以及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到同樣結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,由木質素可游離相同程度之氫離子。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.
且,取代紫外光使用太陽光時,重複上述製程後,得到同樣結果。Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained after repeating the above process.
使用木質素分解用觸媒,自木質素游離氫離子之方法中,作為木質素使用未含有還原糖纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。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.
將含有上述木質素:2.5mg/mL、2.5mM KOH、及卟啉(Sigmal公司製之原卟啉IV,目錄號碼258385-1G):50μg/mL之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。其結果得知木質素反應液的pH自10.7降至7.4。因此,得知由木質素游離氫離子。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.
又,取代紫外光使用太陽光,重複上述製程後,得到同樣結果。Further, the use of sunlight instead of ultraviolet light, after repeating the above process, gave the same result.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有該木質素:2.5mg/mL、0.05M KOH、及依據上述製造例1所得之卟啉:50μg/mL之溶液1mL,放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射12小時。將所得之樣品以HPLC使用膠體過濾管柱進行分析。作為對照,對於紫外線照射前之溶液亦進行分析。分析結果得知,照射後之樣品與照射前之樣品相比較,於310nm之吸光中,由在7min所溶離之木質素的波峰面積,72%之木質素被分解。該分解生成物經膠體過濾,溶離出較低分子量的分餾物。因本發明中之木質素的分解生成物之吸光度較低,故在310nm之吸光度無法描繪,作為非揮發性之分解生成物,將溶離樣品進行冷凍乾燥後,收集相同部分得到分解生成物之沈澱。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. .
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、以及不溶解於水之製品(Aldrich公司製,目錄No.370967),實施上述製程後,得到同程度之結果。即,得知與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,於木質素-鹼化合物溶液進一步添加作為光觸媒之卟啉時,可得到與木質素之光分解相關的相同程度之結果。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.
又,取代紫外光使用太陽光,重複上述製程後,得到同樣結果。Further, the use of sunlight instead of ultraviolet light, after repeating the above process, gave the same result.
作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Sigmal公司製,目錄No.471003,分子量60,000)。將含有對於該木質素205mg/mL,0.05M KOH、及作為合成卟啉,分子內含有4個羧基之糞卟啉I(ALDRICH公司製)50μg/mL的溶液0.5mL,使其殘留空氣空間下,將2mL容量放於持有聚丙烯製Eppendorf tube程度之光透過性的圓筒形試管,使用Spectronix公司製之ENF型紫外光燈以紫外光照射48小時。將如此所得之反應液與實施例2的相同情況下進行處理。其結果確認木質素之80%以上被分解。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.
又,對於空氣之空間以氧氣填充,與上述同樣下以紫外光照射後,同樣地確認出木質素之80%以上被分解。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.
且,對於空氣之空間以氧氣填充,與上述同樣下以紫外光照射後,確認出木質素的10%程度被分解。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.
且,又將上述之含有卟啉的木質素溶液,以Eppendorf tube內不會有空氣空間之量下進行填充,與上述同樣下以紫外光照射後,幾乎未確認到木質素之分解。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.
又,作為木質素使用未含還原糖或纖維素等雜質,純度高之製品(Aldrich公司製,目錄No.471046,分子量12,000)、含有還原糖而純度稍低之製品(Sigmal公司製,目錄No.471038,分子量52,000)、及不溶解於水之製品(Aldrich公司製,目錄No.370967),重複上述製程後,得到對於木質素之分解的同程度結果。即,與所使用之木質素中存在雜質、木質素之平均分子量、或木質素對水之溶解性無關,可將木質素於同程度下進行分解。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.
且,取代紫外光使用太陽光時,重複上述製程後,對於木質素之分解得到同樣結果。Moreover, when sunlight is used instead of ultraviolet light, the same result is obtained for the decomposition of lignin after repeating the above process.
本發明的木質素分解用觸媒並非為白色腐朽菌等酵素,其係由非蛋白質所成之觸媒。藉由使用該觸媒,主要為使用光能量來分解木質素,由木材的僅到達20~30%之非有效資源的木質素,可經簡便方法,游離甲醇等醇類或甲酸等有機酸類或氫離子,故可擴大在自然界為分解困難的木質素之利用領域。即,由木質素所取出之醇類可作為石油等化石燃料的代替燃料,而利用於燃料產業之領域中,有機酸類可利用於各種產業領域中,又由木質素所得之氫離子,例如可利用於燃料電池,故可利用於電力產業之領域中。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.
又,本發明之芳香族烴分解用觸媒為,可分解於苯環結合氧原子之芳香族烴,故可使有害產業廢棄物之處理、或已被污染的土壤或地表的淨化成為可能。因此,可利用於產業廢棄物處理領域、或土壤等淨化領域中。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.
[圖1]製造例1所得之產物的結構式。1 is a structural formula of the product obtained in Production Example 1.
[圖2]對於製造例1所得之產物,表示吸光度分析結果之光譜。2] The product obtained in Production Example 1 shows the spectrum of the result of the absorbance analysis.
[圖3]對於製造例1所得之試料1之1H NMR光譜。Fig. 3 is a 1 H NMR spectrum of Sample 1 obtained in Production Example 1.
[圖4]對於製造例1所得之試料1之13C NMR光譜。4 is a 13 C NMR spectrum of Sample 1 obtained in Production Example 1.
[圖5]對於製造例1所得之試料1之HSQC光譜。Fig. 5 is a view showing the HSQC spectrum of Sample 1 obtained in Production Example 1.
[圖6]對於製造例1所得之試料1之COSY光譜。Fig. 6 is a COSY spectrum of Sample 1 obtained in Production Example 1.
[圖7]對於製造例1所得之試料1之HMBC光譜。Fig. 7 is a HMBC spectrum of Sample 1 obtained in Production Example 1.
[圖8]對於製造例1所得之試料1之NOESY光譜。8] The NOESY spectrum of the sample 1 obtained in Production Example 1.
[圖9]對於製造例1所得之試料1之NOESY光譜。9 shows the NOESY spectrum of Sample 1 obtained in Production Example 1.
[圖10]對於製造例1所得之試料2之1H NMR光譜。Fig. 10 is a 1 H NMR spectrum of Sample 2 obtained in Production Example 1.
[圖11]對於製造例1所得之試料2之13C NMR光譜。Fig. 11 is a 13 C NMR spectrum of Sample 2 obtained in Production Example 1.
[圖12]將對於製造例1所得之試料2之NEOSY光譜擴大表示。Fig. 12 is an enlarged view showing the NEOSY spectrum of Sample 2 obtained in Production Example 1.
[圖13]表示對於製造例1所得之試料2之相對存在量的光譜。Fig. 13 is a graph showing the relative amount of the sample 2 obtained in Production Example 1.
[圖14]表示以ESI-MS進行分析之結果。Fig. 14 shows the results of analysis by ESI-MS.
[圖15]表示將製造例1所得之產物以熱分解GC-MS進行分析之結果。Fig. 15 shows the results of analysis of the product obtained in Production Example 1 by thermal decomposition GC-MS.
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