JP3787002B2 - Method for producing a rutinose glycoside of a phenol compound - Google Patents

Description

【００１３】
（ここで、ＲＯＨはフェノール化合物である。）
本発明におけるフェノール化合物は、上記の酵素反応においてルチノースの受容体として作用するものであれば特に限定されない。しかし、好ましいフェノール化合物としては、ヒドロキノン、ピロガロール、ベラトリルアルコール、没食子酸、カテコール、エラグ酸、ベンジルアルコール等が挙げられる。
【００１４】
本発明で用いるルチンは、上記式に示されるように、アグリコンであるクエルセチンに、ルチノース（グルコースとラムノースとからなる二糖）がＯ- グリコシド結合した配糖体である。このルチンは、エンジュ（豆科）、蕎麦（タデ科）、ヘンルーダ（ミカン科）等の天然の植物に広く分布するフラボノール配糖体の一種であり、動脈硬化、高血圧の予防等に有用な生理活性物質として知られている。
【００１５】
本発明におけるルチン分解酵素としては、ルチンを分解してルチノースを切り出し、これをフェノール化合物の酸素原子上に転移させ得るものであれば如何なるものでもよい。その例としては、エンジュ、蕎麦、ヘンルーダ等の植物に由来する酵素、或いはペニシリウム(Penicillium) 属、アスペルギルス(Aspergillus) 属の菌などの微生物に由来する酵素を挙げることができる。これらの酵素のうち、特に好ましいのはダッタン蕎麦の種実に含まれるルチン分解酵素である。何故なら、このルチン分解酵素は活性が高く、またダッタン蕎麦は現在も広く食用に供されているものであり、安全性の点で信頼性が高いからである。また、アスペルギルス・ニガー(Aspergillus niger) から得たルチン分解酵素は、広範囲のフェノール化合物に対して配糖化能を有する点で好ましい。
【００１６】
本発明において、上記のルチン分解酵素は精製品であることが好ましいが、その活性の強さや含量によっては、必ずしも精製品である必要はない。即ち、これらの酵素を含有する植物や微生物の抽出液をそのまま用いたり、場合によっては植物体や微生物をそのまま用いてもよい。
【００１７】
ダッタン蕎麦の種実に含まれるルチン分解酵素を例にとると、ダッタン蕎麦の種実、またはこの種実を挽いて調製した蕎麦粉にバッファーを加えて攪拌抽出をして抽出液を得、これを更に精製して得られた精製酵素を用いることが好ましい。しかし、ダッタン蕎麦に含まれる酵素は活性が高いので、抽出液をそのまま用いても十分に機能する。更に、ダッタン蕎麦の種実には、通常の蕎麦の種実に比較して非常に多量のルチンが含まれている。即ち、通常の蕎麦の種実では約１４ｍｇ％であるのに対して、ダッタン蕎麦の種実では約１３００ｍｇ％（ｍｇ％は、蕎麦種実１００ｇに含まれるルチンのｍｇ量を表す）である。従って、ダッタン蕎麦の種実またはその粉をそのまま用いても、十分に目的とするルチノースの転移反応を行うことができる。
【００１８】
また、微生物に由来するルチン分解酵素を使用する際には、微生物そのものから酵素を抽出して使用してもよい。しかし、その微生物が菌体外に酵素を分泌する場合には、微生物を培養した培養液をそのまま粗酵素液として用いることができる。勿論、この培養液を更に抽出精製し、精製酵素にして用いることも可能である。
【００１９】
植物や微生物からのルチン分解酵素の抽出は、通常、酵素の抽出に用いられる方法により行うことができる。例えば、酢酸バッファー中で攪拌して抽出すればよい。
【００２０】
本発明の方法によれば、一段階の反応で、フェノール化合物をそのルチノース配糖体に変換することができる。従って、精製したルチン分解酵素は、そのままルチンに加えて反応させることもできるが、適当な担体に固定してバイオリアクターを構成すれば、連続的に目的の配糖体を製造することが可能となり、効率を飛躍的に向上させることができる。
【００２１】
本発明によるルチノース配糖体の製造方法は、ルチン分解酵素が失活しない限り、如何なる条件下でも実施することができる。しかし、ｐＨ３〜９、温度８０℃以下で行うことが好ましく、ｐＨ５、温度４０℃で行うのが最も好ましい。
【００２２】
【実施例】
＜実施例１＞
（１）粗酵素液の調製
ダッタン蕎麦粉５０ｇに２０ｍＭ酢酸バッファー（ｐＨ５）を１．５リットル加え、１時間攪拌抽出した後、東洋濾紙製Ｎｏ．１濾紙で濾過することにより、粗酵素液１．４１リットルを得た。
【００２３】
（２）配糖体の製造
ルチン４ｇと、ヒドロキノン４００ｍｇと、上記で得た粗酵素液２０ｍｌとを混合し、４０℃で２４時間攪拌して反応させた。次いで、２０ｍｌの水を加えて１００００ｒｐｍで遠心分離し、濾過した。続いて、濾液を活性炭カラムにかけた後、１０％エタノールで洗浄することにより、ルチノースを除去した。更に、エタノール１００％で溶出させ、減圧留去した後、酢酸エチルを加えて洗浄してヒドロキノンを除去し、その後下記の条件でＨＰＬＣにかけて、２４．７ｍｇの精製物を得た。
【００２４】
カラム；Ａｓａｈｉｐａｋ ＮＨ₂ Ｐ−５０ ４．６＊２５０ｍｍ
（昭和電工株式会社製）
検出器；ＲＩ検出器、ＵＶ検出器
溶離液；アセトニトリル：水＝７５：２５
流速 ；１．０ｍｌ／ｍｉｎ
上記で得られた精製物をＨＰＬＣ分析した結果を、図１および図２に示す。図１はＵＶ検出器による結果であり、図２はＲＩ検出器による結果である。
【００２５】
また、上記で得た精製物のＵＶ吸収スペクトルを図３に、またヒドロキノンのＵＶ吸収スペクトルを図４に示す。
更に、上記で得た精製物の¹³Ｃ−ＮＭＲ分析を行ったところ、図５に示す結果が得られた。
【００２６】
これらの結果から、上記の反応によって得られた生成物は、下記の式で表されるヒドロキノンのルチノース配糖体（ヒドロキノンルチノシド：以下、ＨＱＲと称する）であることが分かった。
【００２７】
【化２】

【００２８】
実施例２
ダッタン蕎麦の代わりにエンジュを用い、それ以外は全て実施例と同様に行った。その結果、６．５ｍｇの精製ＨＱＲが得られた。
【００２９】
実施例３： ＨＱＲの酸分解
実施例１、２で得た精製物（ＨＱＲ）１ｍｇを、２Ｎ塩酸溶液１．２ｍｌ中に溶解し、ブロックヒータで１００℃に加熱して、２時間分解反応を行った。その後、エタノールを添加しながら減圧留去した。得られた分解物について、実施例１で行ったのと同様のＨＰＬＣ分析を行なった。その結果を、図６（ＵＶ検出器）および図７（ＲＩ検出器）に示す。
【００３０】
この結果も、実施例１、２で得た精製物がＨＱＲであることを支持している。
実施例４
上記で得られた精製ＨＱＲのチロシナーゼ阻害活性を調べるために、下記の試験溶液を調製した。
【００３１】

なお、チロシナーゼとしては、マッシュルーム起源のチロシナーゼ（和光純薬株式会社製）を用いた。
【００３２】
上記の試験液および対照溶液を２５℃でインキュベートし、３０分毎に４７０ｎｍでの吸光度を測定した。その結果を図８に示す。
図８の結果から、ＨＱＲはチロシナーゼの活性を阻害し、チロシンの分解を抑制することが分かった。
【００３３】
実施例５
ＩＦ０から分譲を受けたアスペルギルス・ニガー(Aspergillus niger) を、２％ルチンを含む液体培地中で、３０℃で８日間の培養を行い、その培養液を回収した。
【００３４】
この培養液をそのままルチン分解酵素として用い、実施例１に類似した方法で、ルチンとヒドロキノンとの反応を行った。即ち、ルチン、ヒドロキノンおよび上記の培養濾液を混合し、３０℃で２日間反応させた。遠心分離および吸引濾過により、未反応のルチンおよび生成したケルセチン等を沈殿として除去した。そのときの上清を回収し、これに酢酸エチルを加え、未反応のヒドロキノンを上層として除去した。一方、下層を活性炭クロマトグラフィーに供試、その３０％１−プロパノールによる溶出画分を回収した。
【００３５】
こうして得られた生成物の¹³Ｃ−ＮＭＲスペクトルは、図５に示したものと同じであった。このＮＭＲデータから、生成物は実施例１と同じくＨＱＲであることが分かった。また、¹ Ｈ−ＮＭＲスペクトル（データは省略）から、そのアノマー構造はβであることが分かった。
【００３６】
また、ヒドロキノンの代わりに、ピロガロール、ベラトリルアルコール、没食子酸を用い、上記と同様の反応および精製を行った。その結果、ヒドロキノン以外のこれらフェノール化合物についても、同様にルチノース配糖体の生成が認められた。
【００３７】
実施例６： ＨＱＲの抗酸化能
上記の実施例で得たヒドロキノンルチノシド（ＨＱＲ）の抗酸化性を調べるために、高速液体クロマトグラフィー（ＨＰＬＣ）／電気化学検出器（ＥＣＤ）による還元性の測定を行った。電気化学検出器は、電極間に一定の電位を印加して、そこを通る試料による電流の変化を測定するものである。電極間に印加する電位を正の電位とすれば、この条件で検出される物質は、還元力、即ち抗酸化能を有しているということができる。
【００３８】
５ｍＭのＨＱＲを高速液体クロマトグラフィー（カラム：ＹＭＣ社製のＹＭＣ−Ｐａｃｋ ＮＨ₂ ２５０×４．６ｍｍ Ｉ．Ｄ．、溶離液：４０ｍＭ酢酸アンモニウム、１７．５ｍＭ酢酸／８０％メタノール、流速：１ｍｌ／ｍｉｎ）に２０μｌ注入し、溶出してくる試料を電気化学検出器（資生堂社製アンペロメトリック式電気化学検出器、印加電圧＋８００ｍＶ）により検出した。
【００３９】
比較のために、抗酸化活性を有するとされているアルブチンおよびコウジ酸の同濃度溶液を同様の方法で測定した。
この方法によって測定した各物質の電流変化値（ピーク高さから算出）は、以下の通りであった。
【００４０】

上記の結果から、ＨＱＲはアルブチンと略同等、コウジ酸よりも３〜４倍強い還元力を有することが分かり、ＨＱＲが抗酸化能を有していることが示された。
【００４１】
【発明の効果】
以上詳述したように、本発明によれば、フェノール化合物から、簡便かつ効率的に、そのルチノース配糖体を製造することができる。また、本発明により得られるフェノール化合物のルチノース配糖体は、糖部分が二糖のルチノースであるため、新規化合物である。また、本発明の方法により得られる配糖体は、チロシナーゼ阻害活性、美白効果、メラニン生成抑制、活性酸素抑制といった有用な薬理作用を有し、かつ副作用や難溶性といった欠点も少ないものが多い。従って、本発明は斯かる有用な化合物を提供するものとして、社会に裨益するところ大なるものである。
【図面の簡単な説明】
【図１】実施例１で得られた精製物（ＨＱＲ）のＨＰＬＣ分析（ＵＶ検出器）の結果を示す図である。
【図２】実施例１で得られた精製物（ＨＱＲ）のＨＰＬＣ分析（ＲＩ検出器）の結果を示す図である。
【図３】実施例１で得た精製物（ＨＱＲ）のＵＶ吸収スペクトルを示す図である。
【図４】ヒドロキノンのＵＶ吸収スペクトルを示す図である。
【図５】実施例１で得られた精製物（ＨＱＲ）の¹³ＣＮＭＲスペクトルを示す図である。
【図６】実施例３で得たＨＱＲの分解生成物のＨＰＬＣ分析（ＵＶ検出器）の結果を示す図である。
【図７】実施例３で得たＨＱＲの分解生成物のＨＰＬＣ分析（ＲＩ検出器）の結果を示す図である。
【図８】ＨＱＲのチロシナーゼ阻害活性を示す図である。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a phenol compound glycoside having useful functions such as UV absorption ability, antioxidant ability, whitening effect, and tyrosinase inhibitory activity, and more particularly to a production method for producing a rutinose glycoside of a phenol compound.
[0002]
[Prior art]
The glycosides of phenolic compounds are widely present in plants even in small amounts, and have low physiological toxicity despite their excellent physiological activity. Therefore, development of food materials, food additives, pharmaceuticals, cosmetics, etc. Is attracting attention as a target.
[0003]
For example, arbutin is a type of conventionally known phenol compound glycoside. Its chemical name is hydroquinone β-D-glucopyranoside, which is a glucose glycoside having hydroquinone as an aglycone. Arbutin is widely used as a cosmetic because it has whitening effects, melanin production inhibition and active oxygen inhibition. In addition, glycosides containing catechol, resorcinol and the like as aglycones are also known to be effective in preventing and treating skin pigmentation.
[0004]
When the glycosides of phenolic compounds having excellent pharmacological effects as described above are examined in more detail, the aglycones of each glycoside, that is, hydroquinone, catechol, and resorcinol itself are excellent for each glycoside. Have pharmacological effects. However, the effects of these aglycones themselves are small compared to the respective glycosides. Therefore, it is not only forced to be used at a high concentration, but also has side effects such as vitiligo on the skin when used over a long period of time.
[0005]
From the above facts, it is expected that a phenol compound having a useful pharmacological effect generally improves its pharmacological effect and reduces side effects by making it a glycoside. Furthermore, it is known that glycosides are generally more water soluble than the aglycone itself.
[0006]
On the other hand, for example, ellagic acid, which is a dimer of gallic acid, has excellent pharmacological effects such as an antioxidant action and a melanin inhibitory action, but is easily oxidized and unstable, and is soluble in water. Because of its extremely low nature, there is a problem that it is inconvenient to handle. Therefore, with respect to such a phenol compound, it is expected that the problem of side effects and solubility can be improved by making this a glycoside without impairing its physiological activity.
[0007]
However, because there is no efficient and simple method for converting phenolic compounds to glycosides, certain phenolic compounds are not fully utilized despite having useful activity. This is the actual situation.
[0008]
Moreover, the sugar part in the conventional phenol glycoside is limited to several kinds, such as glucose and ribose, and the combination of an aglycone and sugar was comparatively limited. Therefore, even if the same aglycone is used, if a different sugar can be bound, it is expected that a useful novel phenol glycoside can be obtained.
[0009]
[Problems to be solved by the invention]
This invention is made | formed in view of the said situation, and it aims at providing the novel method for manufacturing the rutinose glycoside from a phenol compound.
Moreover, this invention provides the rutinose glycoside of the novel phenol compound obtained by said method.
[0010]
[Means for Solving the Problems]
The present invention relates to a method for producing a rutinose glycoside of a phenol compound, which comprises reacting a rutin-degrading enzyme with rutin in the presence of the phenol compound, and a novel phenol compound rutinose obtained by this method. It is a glycoside.
[0011]
DETAILED DESCRIPTION OF THE INVENTION
The method according to the present invention is to transfer a sugar moiety contained in rutin, that is, rutinose, to the phenol compound by causing a rutin degrading enzyme to act on rutin in the presence of a phenol compound. expressed.
[0012]
[Chemical 1]

[0013]
(Here, ROH is a phenol compound.)
The phenol compound in the present invention is not particularly limited as long as it acts as a receptor for rutinose in the above enzyme reaction. However, preferred phenolic compounds include hydroquinone, pyrogallol, veratryl alcohol, gallic acid, catechol, ellagic acid, benzyl alcohol and the like.
[0014]
Rutin used in the present invention is a glycoside in which rutinose (a disaccharide composed of glucose and rhamnose) is O-glycosidically bonded to quercetin, which is an aglycon, as shown in the above formula. This rutin is a kind of flavonol glycoside widely distributed in natural plants such as Enju (legume), buckwheat (Tapaceae), and Henruda (Rutaceae), and is useful for preventing arteriosclerosis and hypertension. Known as active substance.
[0015]
The rutin-degrading enzyme in the present invention may be any as long as it can decompose rutin to cut out rutinose and transfer it to the oxygen atom of the phenol compound. Examples thereof include enzymes derived from plants such as Enju, oats, and Henruda, or enzymes derived from microorganisms such as Penicillium and Aspergillus bacteria. Among these enzymes, particularly preferred is a rutin-degrading enzyme contained in the seeds of dartane buckwheat. This is because this rutin-degrading enzyme has high activity, and tartane buckwheat is still widely used for food and is highly reliable in terms of safety. In addition, a rutin degrading enzyme obtained from Aspergillus niger is preferable in that it has glycosylation ability for a wide range of phenolic compounds.
[0016]
In the present invention, the rutin-degrading enzyme is preferably a purified product, but depending on the strength and content of its activity, it is not necessarily a purified product. That is, plant or microorganism extracts containing these enzymes may be used as they are, or in some cases, plant bodies or microorganisms may be used as they are.
[0017]
Taking rutin-degrading enzyme contained in the seeds of tartane buckwheat as an example, buffer is added to the seeds of tartnut soba seeds or buckwheat flour prepared by grinding these seeds to obtain an extract, which is further purified. It is preferable to use the purified enzyme obtained in this way. However, since the enzyme contained in tartane buckwheat has high activity, it functions well even if the extract is used as it is. Furthermore, the seeds of tartane buckwheat contain a very large amount of rutin compared to the seeds of normal buckwheat. That is, it is about 14 mg% in the normal seed of buckwheat noodles, whereas it is about 1300 mg% in the seed of buckwheat noodles (mg% represents the amount of rutin contained in 100 g of buckwheat seeds). Therefore, even if the seed seeds of tartane buckwheat or its flour are used as they are, the intended transfer reaction of rutinose can be sufficiently performed.
[0018]
Further, when using a rutin-degrading enzyme derived from a microorganism, the enzyme may be extracted from the microorganism itself. However, when the microorganism secretes the enzyme outside the cells, the culture solution in which the microorganism is cultured can be used as a crude enzyme solution as it is. Of course, it is also possible to further extract and purify this culture solution and use it as a purified enzyme.
[0019]
Extraction of rutin-degrading enzyme from plants and microorganisms can be usually performed by a method used for enzyme extraction. For example, the extraction may be performed by stirring in an acetate buffer.
[0020]
According to the method of the present invention, a phenol compound can be converted into its rutinose glycoside in a single step reaction. Therefore, the purified rutin-degrading enzyme can be reacted by adding it directly to rutin. However, if the bioreactor is constructed by fixing it to an appropriate carrier, it is possible to continuously produce the desired glycoside. , Efficiency can be improved dramatically.
[0021]
The method for producing a rutinose glycoside according to the present invention can be carried out under any conditions as long as the rutin degrading enzyme is not inactivated. However, it is preferably performed at a pH of 3 to 9 and a temperature of 80 ° C. or less, and most preferably at a pH of 5 and a temperature of 40 ° C.
[0022]
【Example】
<Example 1>
(1) Preparation of crude enzyme solution After adding 1.5 liters of 20 mM acetate buffer (pH 5) to 50 g of tartane buckwheat flour, the mixture was stirred and extracted for 1 hour. By filtering with 1 filter paper, 1.41 liters of crude enzyme solution was obtained.
[0023]
(2) Production of Glycoside 4 g of rutin, 400 mg of hydroquinone and 20 ml of the crude enzyme solution obtained above were mixed and reacted by stirring at 40 ° C. for 24 hours. Next, 20 ml of water was added and centrifuged at 10,000 rpm and filtered. Subsequently, the filtrate was applied to an activated carbon column and then washed with 10% ethanol to remove rutinose. Furthermore, after eluting with 100% ethanol and distilling off under reduced pressure, ethyl acetate was added and washed to remove hydroquinone, and then subjected to HPLC under the following conditions to obtain 24.7 mg of a purified product.
[0024]
Column; Asahipak NH ₂ P-50 4.6 * 250 mm
(Showa Denko Co., Ltd.)
Detector; RI detector, UV detector eluent; Acetonitrile: water = 75: 25
Flow rate: 1.0 ml / min
The results of HPLC analysis of the purified product obtained above are shown in FIG. 1 and FIG. FIG. 1 shows the result of the UV detector, and FIG. 2 shows the result of the RI detector.
[0025]
Further, the UV absorption spectrum of the purified product obtained above is shown in FIG. 3, and the UV absorption spectrum of hydroquinone is shown in FIG.
Furthermore, when the purified product obtained above was subjected to ¹³ C-NMR analysis, the results shown in FIG. 5 were obtained.
[0026]
From these results, it was found that the product obtained by the above reaction was a hydroquinone rutinose glycoside represented by the following formula (hydroquinone rutinoside: hereinafter referred to as HQR).
[0027]
[Chemical 2]

[0028]
Example 2
Enju was used instead of tartane soba, and everything else was performed in the same manner as in the example. As a result, 6.5 mg of purified HQR was obtained.
[0029]
Example 3 Acid Decomposition of HQR 1 mg of the purified product (HQR) obtained in Examples 1 and 2 was dissolved in 1.2 ml of 2N hydrochloric acid solution, heated to 100 ° C. with a block heater, and decomposed for 2 hours. went. Then, it distilled off under reduced pressure, adding ethanol. The obtained decomposition product was subjected to the same HPLC analysis as that performed in Example 1. The results are shown in FIG. 6 (UV detector) and FIG. 7 (RI detector).
[0030]
This result also supports that the purified product obtained in Examples 1 and 2 is HQR.
Example 4
In order to examine the tyrosinase inhibitory activity of the purified HQR obtained above, the following test solution was prepared.
[0031]

As tyrosinase, mushroom-derived tyrosinase (manufactured by Wako Pure Chemical Industries, Ltd.) was used.
[0032]
The above test solution and control solution were incubated at 25 ° C., and the absorbance at 470 nm was measured every 30 minutes. The result is shown in FIG.
From the results of FIG. 8, it was found that HQR inhibits tyrosinase activity and suppresses tyrosine degradation.
[0033]
Example 5
Aspergillus niger received from IF0 was cultured at 30 ° C. for 8 days in a liquid medium containing 2% rutin, and the culture solution was collected.
[0034]
This culture solution was directly used as a rutin-degrading enzyme, and a reaction between rutin and hydroquinone was carried out in the same manner as in Example 1. That is, rutin, hydroquinone and the above culture filtrate were mixed and reacted at 30 ° C. for 2 days. Unreacted rutin, produced quercetin and the like were removed as precipitates by centrifugation and suction filtration. The supernatant at that time was collected, ethyl acetate was added thereto, and unreacted hydroquinone was removed as an upper layer. On the other hand, the lower layer was subjected to activated carbon chromatography, and the fraction eluted with 30% 1-propanol was collected.
[0035]
The ¹³ C-NMR spectrum of the product thus obtained was the same as that shown in FIG. From this NMR data, it was found that the product was HQR as in Example 1. From the ¹ H-NMR spectrum (data not shown), the anomeric structure was found to be β.
[0036]
Further, pyrogallol, veratryl alcohol, and gallic acid were used in place of hydroquinone, and the same reaction and purification as described above were performed. As a result, the production of rutinose glycosides was similarly observed for these phenol compounds other than hydroquinone.
[0037]
Example 6: Antioxidant ability of HQR In order to investigate the antioxidant property of hydroquinone rutinoside (HQR) obtained in the above example, the reductive property by high performance liquid chromatography (HPLC) / electrochemical detector (ECD) Measurements were made. The electrochemical detector applies a constant potential between electrodes and measures a change in current caused by a sample passing therethrough. If the potential applied between the electrodes is a positive potential, it can be said that the substance detected under this condition has a reducing power, that is, an antioxidant ability.
[0038]
5 mM HQR was subjected to high performance liquid chromatography (column: YMC-Pack NH ₂ 250 × 4.6 mm ID manufactured by YMC, eluent: 40 mM ammonium acetate, 17.5 mM acetic acid / 80% methanol, flow rate: 1 ml / Min) was injected at 20 μl, and the eluted sample was detected by an electrochemical detector (amperometric electrochemical detector manufactured by Shiseido Co., Ltd., applied voltage +800 mV).
[0039]
For comparison, the same concentration solution of arbutin and kojic acid, which are said to have antioxidant activity, was measured by the same method.
The current change value (calculated from the peak height) of each substance measured by this method was as follows.
[0040]

From the above results, it was found that HQR is approximately the same as arbutin and has a reducing power 3 to 4 times stronger than kojic acid, indicating that HQR has antioxidant ability.
[0041]
【The invention's effect】
As described above in detail, according to the present invention, the rutinose glycoside can be produced from a phenol compound simply and efficiently. Moreover, the rutinose glycoside of the phenol compound obtained by the present invention is a novel compound because the sugar moiety is a disaccharide rutinose. In addition, many glycosides obtained by the method of the present invention have useful pharmacological actions such as tyrosinase inhibitory activity, whitening effect, melanin production inhibition, and active oxygen inhibition, and have few side effects and poor solubility. Therefore, the present invention greatly contributes to society as providing such useful compounds.
[Brief description of the drawings]
1 is a diagram showing the results of HPLC analysis (UV detector) of the purified product (HQR) obtained in Example 1. FIG.
2 is a diagram showing the results of HPLC analysis (RI detector) of the purified product (HQR) obtained in Example 1. FIG.
3 is a diagram showing a UV absorption spectrum of the purified product (HQR) obtained in Example 1. FIG.
FIG. 4 is a diagram showing the UV absorption spectrum of hydroquinone.
5 is a diagram showing a ¹³ C NMR spectrum of the purified product (HQR) obtained in Example 1. FIG.
6 is a diagram showing the results of HPLC analysis (UV detector) of the decomposition product of HQR obtained in Example 3. FIG.
7 is a view showing the results of HPLC analysis (RI detector) of the HQR decomposition product obtained in Example 3. FIG.
FIG. 8 is a graph showing tyrosinase inhibitory activity of HQR.

Claims (6)

  In the presence of a phenolic compound selected from the group consisting of hydroquinone, pyrogallol and gallic acid, rutinase is allowed to act on rutin, whereby the phenolic compound is its phenolic hydroxyl group at the 1st position on the glucose side of rutinose. A method for producing a rutinose glycoside of a phenol compound, characterized by obtaining a rutinose glycoside having a glycosidic bond to carbon.   The method for producing a rutinose glycoside according to claim 1, wherein the phenol compound is hydroquinone.   A rutinose glycoside in which a phenol compound selected from the group consisting of hydroquinone, pyrogallol and gallic acid is glycoside-bonded to the first carbon on the glucose side of rutinose at its phenolic hydroxyl group.   The rutinose glycoside according to claim 3, wherein the phenol compound is hydroquinone.   An active oxygen inhibitor comprising, as an active ingredient, a rutinose glycoside in which a phenol compound selected from the group consisting of hydroquinone, pyrogallol, and gallic acid is glycosylated with a phenolic hydroxyl group to the first carbon on the glucose side of rutinose.   The active oxygen inhibitor according to claim 5, wherein the phenol compound is hydroquinone.

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