JP2013127027A - Ultraviolet absorber composition originated from plant inhabiting microbes - Google Patents
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- XNEFYCZVKIDDMS-UHFFFAOYSA-N avobenzone Chemical compound C1=CC(OC)=CC=C1C(=O)CC(=O)C1=CC=C(C(C)(C)C)C=C1 XNEFYCZVKIDDMS-UHFFFAOYSA-N 0.000 description 3
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- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 1
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
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- 230000008845 photoaging Effects 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
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- OGIDPMRJRNCKJF-UHFFFAOYSA-N titanium oxide Inorganic materials [Ti]=O OGIDPMRJRNCKJF-UHFFFAOYSA-N 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Cosmetics (AREA)
Abstract
Description
本発明は、農作物等の植物表面に生息する微生物に由来する紫外線吸収剤組成物及びその製造法に関する。 The present invention relates to an ultraviolet absorber composition derived from a microorganism that inhabits the surface of a plant such as an agricultural crop and a method for producing the same.
紫外線防止剤は、化粧品や日焼け止め製品、その他工業製品において、皮膚や製品を紫外線から守るために従来より様々な種類のものが使用されている。例えば化粧品や日焼け止め製品に使用される紫外線防止剤は、主にUV−A(320-400nmの長波長紫外線)とUV−B(280-320nmの中波長紫外線)に様々な観点から対応するものであり、より具体的には、反射剤(酸化チタン、酸化亜鉛等)、吸収剤(オキシベンゾン、アボベンゾン等)、抗酸化剤(ビタミン、カロチノイド等)などが使用されている。 Various types of UV inhibitors have been used in cosmetics, sunscreen products, and other industrial products in order to protect the skin and products from UV rays. For example, UV inhibitors used in cosmetics and sunscreen products mainly correspond to UV-A (320-400 nm long wavelength UV) and UV-B (280-320 nm medium wavelength UV) from various viewpoints. More specifically, reflective agents (such as titanium oxide and zinc oxide), absorbents (such as oxybenzone and avobenzone), and antioxidants (such as vitamins and carotenoids) are used.
上記のうち、素材としてほぼ確立している反射剤、既に多様な種類のものが存在する抗酸化剤と比較して、紫外線吸収剤はまだ実用されている種類が少ない。また、化粧品原料等に用いられる既存の紫外線波長UV−Aの吸収剤のオキシベンゾンやアボベンゾン等は化学合成品であるが、これらの物質は炎症や肌荒れの原因になる場合もあるため、皮膚に処理するという観点上、天然由来成分の物質に代替されることが市場で望まれている。また、UV−Aが皮膚の光老化の主因であるとする認識が広まり、UV−A対策に関する消費者の関心も高い。 Among the above, there are few types of UV absorbers that are still in practical use, compared to reflective agents that are almost established as materials and antioxidants that already have various types. In addition, oxybenzone and avobenzone, which are existing UV-A UV absorbers used as cosmetic raw materials, are chemically synthesized products, but these substances may cause irritation and rough skin. In view of the above, it is desired in the market to be replaced by a substance derived from a natural component. Moreover, the recognition that UV-A is the main cause of photoaging of the skin is widespread, and consumers are highly interested in UV-A countermeasures.
天然由来の紫外線吸収物質として、海洋に生息する細菌やラン藻類の生産するマイコスポリン様化合物やサイトネミン等がこれまでに報告されている(特許文献1,2;非特許文献1、2、3、4)。しかしながら、地上の農作物体上に普遍的に生息する微生物に着眼して発見された例はなく、それらが含有する紫外線吸収成分については知られていない。また、既知の吸収剤は極性が低い性状を有するものが多く、加工特性の観点からも課題が存在した。 As a naturally occurring ultraviolet absorbing substance, mycosporin-like compounds, cytonemin, etc. produced by marine bacteria and cyanobacteria have been reported so far (Patent Documents 1, 2; Non-Patent Documents 1, 2, 3, 4). ). However, there have been no examples that have been discovered focusing on microorganisms that inhabit universally on the ground crops, and the UV-absorbing components they contain are not known. In addition, many of the known absorbents have properties with low polarity, and there are problems from the viewpoint of processing characteristics.
本発明は、上記の課題を解決する新規の紫外線吸収剤組成物を提供することを目的とする。 An object of this invention is to provide the novel ultraviolet absorber composition which solves said subject.
本願発明者らは、紫外線等の様々な環境ストレスに曝されている植物葉圏の微生物(植物生息微生物)から、紫外線、特にUV−Aの有効な吸収効果を有する紫外線吸収成分が得られることを見出し、本願発明を完成するに至った。 The inventors of the present application are able to obtain an ultraviolet ray absorbing component having an effective absorption effect of ultraviolet rays, particularly UV-A, from plant phytosphere microorganisms (plant-inhabiting microorganisms) exposed to various environmental stresses such as ultraviolet rays. As a result, the present invention has been completed.
即ち、本発明は、植物生息微生物由来の紫外線吸収剤組成物であって、320-400nmの波長領域に紫外線吸収スペクトルの吸収ピークを有する紫外線吸収剤組成物を提供する。
また、本発明は、上記紫外線吸収剤組成物を含む化粧品を提供する。
また、本発明は、320-400nmの波長領域に紫外線吸収スペクトルの吸収ピークを有する紫外線吸収成分を有する植物生息微生物を培養する工程、植物生息微生物の培養物を溶媒で抽出する工程、及び、溶媒から紫外線吸収成分を分離する工程を含む、前記紫外線吸収成分を含む紫外線吸収剤組成物の製造方法を提供する。
That is, the present invention provides an ultraviolet absorber composition derived from plant-inhabiting microorganisms, which has an absorption peak of an ultraviolet absorption spectrum in a wavelength region of 320 to 400 nm.
Moreover, this invention provides the cosmetics containing the said ultraviolet absorber composition.
The present invention also includes a step of culturing a plant-inhabiting microorganism having an ultraviolet-absorbing component having an absorption peak of an ultraviolet absorption spectrum in a wavelength region of 320 to 400 nm, a step of extracting a plant-inhabiting microorganism culture with a solvent, and a solvent. The manufacturing method of the ultraviolet absorber composition containing the said ultraviolet absorption component including the process of isolate | separating an ultraviolet absorption component from the said is provided.
本発明により、天然成分由来の新規の紫外線吸収剤組成物を提供することができる。また、本発明の紫外線吸収剤組成物は極性が高い性状を有するため、加工特性の観点からも化粧品やインク等の工業製品に対する適用性における利点を有する。 According to the present invention, a novel ultraviolet absorber composition derived from natural components can be provided. Moreover, since the ultraviolet absorber composition of the present invention has a high polarity, it has an advantage in applicability to industrial products such as cosmetics and inks from the viewpoint of processing characteristics.
本発明は、植物生息微生物由来の紫外線吸収剤組成物であって、320-400nmの波長領域に紫外線吸収スペクトルの吸収ピークを有する紫外線吸収剤組成物を提供する。
本発明に使用される植物生息微生物は、所望の紫外線吸収能力を有し、また、所望の波長領域の紫外線の吸収を有するものを当業者が適宜選択することが可能である。例えば、植物から単離された一群の微生物について、それらの抽出物を作成し、UV吸収スペクトルを測定することによって、本発明に適した植物生息微生物をルーチン作業により選択することができる。
The present invention provides an ultraviolet absorber composition derived from plant-inhabiting microorganisms, which has an absorption peak of an ultraviolet absorption spectrum in a wavelength region of 320 to 400 nm.
The plant-inhabiting microorganism used in the present invention can be appropriately selected by those skilled in the art to have a desired ultraviolet absorption ability and to absorb ultraviolet rays in a desired wavelength region. For example, for a group of microorganisms isolated from a plant, plant extracts that are suitable for the present invention can be routinely selected by making extracts and measuring UV absorption spectra.
植物生息微生物の種類としては、上述の方法等によって選択されるものであれば特に限定されず、細菌、酵母、カビなどのいずれでもよいが、上記のうち細菌が好ましく、例えば、Rhizobiales(リゾビウム目)、さらにMethylobacteriaceae(メチロバクテリウム科)に属するものが好ましい。そのなかでも、Methylobacterium属の細菌は植物の葉などに多く生息することが知られるものであり、本発明の目的において特に好ましい。
また、本発明の植物生息微生物の単離源となる植物としては、イネ、コムギ、オオムギ、トマト、イチゴ、ニラ、コマツナ、チンゲンサイ、ダイズなどが挙げられるが、これらに限定されない。
The type of plant-inhabiting microorganism is not particularly limited as long as it is selected by the above-described method and the like, and may be any of bacteria, yeast, mold, etc. Among them, bacteria are preferred, for example, Rhizobiales And those belonging to Methylobacteriaceae (Methylobacteriaceae) are preferred. Among them, bacteria belonging to the genus Methylobacterium are known to inhabit a lot of plant leaves and the like, and are particularly preferable for the purpose of the present invention.
In addition, examples of the plant serving as an isolation source of the plant-inhabiting microorganisms of the present invention include, but are not limited to, rice, wheat, barley, tomato, strawberry, leek, komatsuna, dung beetle, and soybean.
本発明の紫外線吸収剤組成物において、植物生息微生物を紫外線吸収剤組成物に使用する態様は当業者が適宜決定することが可能であり、その培養物をそのまま、又は菌体を乾燥して得られた乾燥物を紫外線吸収剤組成物に使用してもよい。あるいは、適宜培養した植物生息微生物を適切な溶媒で抽出した抽出物を紫外線吸収剤組成物に使用してもよい。好ましくは、培養した植物生息微生物を抽出した溶媒からイオン交換クロマトグラフィ等の手段によって紫外線吸収成分を分離し、紫外線吸収剤組成物に配合される。
培養の方法は特に限定されず、液体培養、固体培養のいずれでもよく、微生物の種類等に応じて適宜決定することができる。
In the ultraviolet absorbent composition of the present invention, the mode in which plant-inhabiting microorganisms are used in the ultraviolet absorbent composition can be appropriately determined by those skilled in the art, and the culture can be obtained as it is or by drying the cells. You may use the obtained dried material for a ultraviolet absorber composition. Or you may use for the ultraviolet absorber composition the extract which extracted the plant inhabitant microorganisms cultured suitably with the appropriate solvent. Preferably, the ultraviolet absorbing component is separated from the solvent extracted from the cultured plant-inhabiting microorganisms by means such as ion exchange chromatography and blended into the ultraviolet absorbent composition.
The culture method is not particularly limited, and may be either liquid culture or solid culture, and can be appropriately determined according to the type of microorganism.
また、抽出物を使用する場合、溶媒は、微生物の種類や、吸収剤が使用される最終製品の種類等に応じて当業者が適宜決定することができる。例えば、一般的な溶媒極性の指標として用いられている溶媒極性パラメーターET値(参考文献:Marcus, Y. 1998. The Property of Solvents, Wiley, ISBN: 978-0-471-98369-9)が45.0〜65.0(kcal/mol, 25℃)の溶媒を使用することが可能であり、より好ましくはET値46.0〜64.0、さらに好ましくは、ET値48.0〜63.5の溶媒を使用することができる。
より具体的な溶媒としては、水(ET値63.1)、メタノール(同55.5)、n-ブタノール(同49.7)、2-プロパノール(同48.6)、エタノール(同51.9)が挙げられ、これらを単独又は混合して使用してもよい。例えば、メタノールと水を混合し、50〜80%のメタノールとして使用することができる。
Moreover, when using an extract, the solvent can be suitably determined by those skilled in the art according to the type of microorganism, the type of final product in which the absorbent is used, and the like. For example, the solvent polarity parameter ET value (reference: Marcus, Y. 1998. The Property of Solvents, Wiley, ISBN: 978-0-471-98369-9) used as a general solvent polarity index is 45.0. It is possible to use a solvent of ˜65.0 (kcal / mol, 25 ° C.), more preferably a solvent having an ET value of 46.0 to 64.0, and still more preferably a solvent having an ET value of 48.0 to 63.5.
More specific solvents include water (ET value 63.1), methanol (55.5), n-butanol (49.7), 2-propanol (48.6), and ethanol (51.9). You may mix and use. For example, methanol and water can be mixed and used as 50-80% methanol.
本発明の紫外線吸収剤組成物は、化粧品等に適宜配合することができる。化粧品の種類は特に限定されず、日焼け止めクリーム、化粧用下地、固形又は液体ファンデーション、UV防止用スプレー、乳液、整髪剤、口紅などが挙げられる。 The ultraviolet absorbent composition of the present invention can be appropriately blended in cosmetics and the like. The type of cosmetic is not particularly limited, and examples thereof include sun cream, makeup base, solid or liquid foundation, UV prevention spray, milky lotion, hair conditioner, lipstick and the like.
本発明の紫外線吸収剤組成物はUV−A吸収活性を有し、その吸収スペクトルを分光光度計で測定した場合、320-400nmの波長領域に吸収ピークを有する。本発明の紫外線吸収剤組成物は、好ましくは、紫外線の波長領域における最大吸収ピークを320-400nm、より好ましくは340-380nm、さらに好ましくは355-365nmの波長範囲内に有する。 The ultraviolet absorbent composition of the present invention has UV-A absorption activity, and has an absorption peak in the wavelength region of 320 to 400 nm when the absorption spectrum is measured with a spectrophotometer. The ultraviolet absorbent composition of the present invention preferably has a maximum absorption peak in the ultraviolet wavelength region within the wavelength range of 320 to 400 nm, more preferably 340 to 380 nm, and even more preferably 355 to 365 nm.
さらに、本発明の紫外線吸収剤組成物は、紫外線吸収スペクトル[340-380nmのピーク面積]/[320-400nmのピーク面積]の比が、0.5以上、好ましくは0.55以上、より好ましくは0.60以上である。上記の吸収スペクトル面積は、紫外線吸収ピークを半値幅法やSWIFT II-METHOD等の各種スペクトル解析ソフトの積分プログラム等により面積値に換算することにより求めることができる。
以下、本発明の実施例を示すが、本発明はこれらに限定されるものではない。
Further, the ultraviolet absorbent composition of the present invention has a ratio of ultraviolet absorption spectrum [peak area of 340-380 nm] / [peak area of 320-400 nm] of 0.5 or more, preferably 0.55 or more, more preferably 0.60 or more. is there. The above-mentioned absorption spectrum area can be obtained by converting the ultraviolet absorption peak into an area value by an integration program of various spectrum analysis software such as half width method or SWIFT II-METHOD.
Examples of the present invention will be described below, but the present invention is not limited thereto.
(1)植物生息微生物の選抜
イネ葉鞘、各種植物の花弁、コムギ穂、イチゴ葉、チンゲンサイ葉から分離された細菌計239菌株を供試し、ジャガイモ・ペプトン・グルコース(PPG: ジャガイモ、200g; ペプトン、5g;グルコース、5g;リン酸二ナトリウム、3g;リン酸一カリウム、0.5g;塩化ナトリウム、3g;蒸留水、1000ml)の寒天平板培地上に塗抹後、25℃で5日間培養した。培養後、平板上の菌体を掻き取り、凍結乾燥等により、約10〜45mgの各菌株の菌体乾燥物を得た。菌体乾燥物に50%メタノール液を4ml入れ、5分間の超音波処理後20分間振とう(160rpm)抽出した。振とう後、遠心分離(10000rpm、10分)により抽出液を回収し、さらにこの抽出工程を2回繰り返して、最終的に約12mlの抽出液を得た。得られた抽出液の100ulを96穴のマイクロプレートに入れ、マルチスペクトロフォトメーター(Viento、大日本製薬株式会社製)および解析ソフトKcJunior(大日本製薬株式会社製)を用いて、抽出液の紫外線吸光スペクトルを測定した。測定は、200〜500nmの波長域を10nmの間隔で連続的に測定するスペクトルスキャン法で行った。その結果、W-213株を含む17菌株の抽出液で、いずれも360nm付近に最大吸収ピークをもつ特徴的スペクトルを示すことを見いだした(図1はW213株のスペクトルを示す)。なお、これは既存の化学合成した吸収剤であるアボベンゾンと同等のスペクトルであった。
(1) Selection of plant-inhabiting microorganisms A total of 239 strains of bacteria isolated from rice leaf sheath, petals of various plants, wheat ears, strawberry leaves, and chingensai leaves were tested and potato peptone glucose (PPG: potato, 200 g; peptone, 5 g; glucose, 5 g; disodium phosphate, 3 g; monopotassium phosphate, 0.5 g; sodium chloride, 3 g; distilled water, 1000 ml) were smeared and cultured at 25 ° C. for 5 days. After culturing, the cells on the plate were scraped off, and about 10 to 45 mg of dried cells of each strain was obtained by freeze-drying or the like. 4 ml of a 50% methanol solution was added to the dried microbial cell, and extracted by shaking (160 rpm) for 20 minutes after sonication for 5 minutes. After shaking, the extract was collected by centrifugation (10000 rpm, 10 minutes), and this extraction process was repeated twice to finally obtain about 12 ml of extract. Put 100 ul of the obtained extract in a 96-well microplate, and use the multi-spectrophotometer (Viento, manufactured by Dainippon Pharmaceutical Co., Ltd.) and analysis software KcJunior (produced by Dainippon Pharmaceutical Co., Ltd.) Absorption spectrum was measured. The measurement was performed by a spectrum scan method in which a wavelength range of 200 to 500 nm was continuously measured at intervals of 10 nm. As a result, it was found that the extract of 17 strains including the W-213 strain showed a characteristic spectrum having a maximum absorption peak around 360 nm (FIG. 1 shows the spectrum of the W213 strain). This spectrum was equivalent to that of avobenzone, an existing chemically synthesized absorbent.
これらの17菌株をrRNA遺伝子の塩基配列に基づき同定した結果、いずれもMethylobacterium属細菌と同定した。特徴的なUV吸収スペクトルを示したMethylobacterium sp. W-213株の写真を図2に示す。 As a result of identifying these 17 strains based on the base sequence of the rRNA gene, all of them were identified as Methylobacterium bacteria. A photograph of Methylobacterium sp. W-213 showing a characteristic UV absorption spectrum is shown in FIG.
(2)抽出溶媒
上述(1)と同じ方法で得たMethylobacterium sp.W-213株の乾燥菌体を出発材料とし、(1)と同様の手順で各種抽出溶媒による抽出を行い、最終的に菌体乾燥重量1mgあたり1mlの量になるように抽出液の濃度を調整した。濃度調整した各溶媒抽出液100μlについて、波長360nmにおける吸光度を分光光度計(Ultrospec 2000、ファルマシア製)および解析ソフトSWIFT II-METHOD(ファルマシア製;Wavescanプログラム、1.0nmステップ、スキャンスピードは6200nm/min)により測定した。各抽出溶媒で得られた抽出液の吸光度(360nm)及び溶媒のET値(kcal/mol, 25℃)を以下に示す。
(2) Extraction solvent Using the dried cells of Methylobacterium sp. W-213 obtained by the same method as in (1) above as a starting material, extraction with various extraction solvents is performed in the same procedure as in (1). The concentration of the extract was adjusted to 1 ml per 1 mg of dry cell weight. Absorbance at a wavelength of 360 nm for 100 μl of each solvent-extracted solution was measured using a spectrophotometer (Ultrospec 2000, Pharmacia) and analysis software SWIFT II-METHOD (Pharmacia; Wavescan program, 1.0 nm step, scan speed 6200 nm / min) It was measured by. The absorbance (360 nm) of the extract obtained with each extraction solvent and the ET value (kcal / mol, 25 ° C.) of the solvent are shown below.
以上より、溶媒極性パラメーターET値が48.6〜63.1である溶媒で植物生息微生物を抽出した場合に、360nmにおける吸収が確認されることが分かる。 From the above, it can be seen that absorption at 360 nm is confirmed when plant inhabiting microorganisms are extracted with a solvent having a solvent polarity parameter ET value of 48.6 to 63.1.
(3)植物生息微生物からの、紫外線吸収成分の抽出
Methylobacterium sp. W-213株を用い、PPG液体培地での振とう培養およびPPG寒天平板上での塗抹培養により菌体を得た(図3)。菌体を乾燥して得られた乾燥物8.169 gを約3リットルの80%メタノールで抽出して活性のある抽出産物(1.13g)を得た。その抽出産物を固相カートリッジカラム(PoraPak, Waters製)を用い、カラムに添付された用法に従いメタノール、次いで濃アンモニア:メタノール=5:95液でステップワイズ溶出し、濃アンモニア:メタノール=5:95液画分である81.4mgの黄褐色の塩基性の吸収成分を活性画分として得た(図4)。
(3) Extraction of UV-absorbing components from plant-inhabiting microorganisms
Using Methylobacterium sp. W-213 strain, cells were obtained by shaking culture on PPG liquid medium and smear culture on PPG agar plate (FIG. 3). An active extract (1.13 g) was obtained by extracting 8.169 g of the dried product obtained by drying the cells with about 3 liters of 80% methanol. The extracted product was stepwise eluted with methanol and then concentrated ammonia: methanol = 5: 95 using a solid phase cartridge column (PoraPak, Waters) according to the usage attached to the column, and concentrated ammonia: methanol = 5: 95. A liquid fraction, 81.4 mg of a yellowish brown basic absorption component, was obtained as the active fraction (FIG. 4).
(4)抽出成分のHPLC解析
吸収成分を分離するため、HPLC(Waters製、600 Controller、2414 Detecter)による解析を行った。カラムはAtlantis T3(3μm、 4.6×150mm、 Waters製)を用い、溶出条件は、0→8分まではH2Oを流量0.5ml/分→0.7ml/分のリニアグラジエントで溶出、8→12分までは0.7ml/分の流量でH2O→100%メタノールのリニアグラジエントで溶出、12→30分までは0.7ml/分の流量で100%メタノールによる溶出とした。その結果、大きく5本のピークが検出され、いずれのピークも吸光度360nmの波長に対し最大吸収ピークを示したことから、本吸収成分は少なくとも5種類の吸収物質で構成されることが判明した(図5)。
(4) HPLC analysis of extracted components In order to separate the absorbing components, analysis by HPLC (Waters, 600 Controller, 2414 Detecter) was performed. The column is Atlantis T3 (3μm, 4.6 x 150mm, manufactured by Waters). Elution conditions are 0 to 8 minutes, H 2 O is eluted with a linear gradient of 0.5 ml / min → 0.7 ml / min, 8 → 12 Elution was performed with a linear gradient of H 2 O → 100% methanol at a flow rate of 0.7 ml / min until min, and elution with 100% methanol at a flow rate of 0.7 ml / min until 12 → 30 min. As a result, five large peaks were detected, and all the peaks showed the maximum absorption peak with respect to the wavelength of absorbance of 360 nm. Therefore, it was found that the present absorption component is composed of at least five kinds of absorbing substances ( FIG. 5).
(5)植物生息微生物の培養物を用いた紫外線吸収剤組成物
コムギの穂から分離したコムギの穂およびイチゴの葉からそれぞれ分離したMethylobacterium sp. W-213株、W-182株、および対照としてトマト葉から分離したBacillus sp. No.20株を用い、ポテト・ペプトン・グルコース(PPG)寒天平板上での塗抹培養により、それぞれの菌体を得た。菌体を乾燥して得られた乾燥物を滅菌水で0.1mg/ml濃度に懸濁し、懸濁液(100ul)の紫外線吸光スペクトルおよび320-400nmの波長域におけるピーク面積を分光光度計(Ultrospec 2000、ファルマシア製)および解析ソフトSWIFT II-METHOD(ファルマシア製;Wavescanプログラム、1.0nmステップ、スキャンスピードは6200nm/min)により測定した。その結果、Bacillus sp. No.20株の菌体懸濁液では320-400nmの波長域での吸収ピークは確認されなかったのに対し、Methylobacterium sp. W-213株、W-182株では確認され、そのピーク面積はそれぞれ、205.9および204.4であった(図6)。
(5) UV absorber composition using a culture of plant-inhabiting microorganisms Methylobacterium sp. W-213 and W-182 strains isolated from wheat ears and strawberry leaves isolated from wheat ears, and as controls Using Bacillus sp. No. 20 strain isolated from tomato leaves, each cell was obtained by smear culture on a potato, peptone, glucose (PPG) agar plate. The dried product obtained by drying the cells is suspended in sterile water at a concentration of 0.1 mg / ml, and the UV absorption spectrum of the suspension (100 ul) and the peak area in the wavelength range of 320-400 nm are measured with a spectrophotometer (Ultrospec 2000, manufactured by Pharmacia) and analysis software SWIFT II-METHOD (Pharmacia; Wavescan program, 1.0 nm step, scan speed 6200 nm / min). As a result, absorption peaks in the wavelength range of 320-400 nm were not confirmed in the cell suspension of Bacillus sp. No. 20, whereas it was confirmed in Methylobacterium sp. W-213 and W-182. The peak areas were 205.9 and 204.4, respectively (FIG. 6).
(6)市販の化粧品と本発明の紫外線吸収剤組成物の吸収スペクトルの相違
上述(1)と同じ方法で得たMethylobacterium sp.W-213株の乾燥菌体および市販の日焼け止めクリームA及びBの乾燥物を出発材料とし、(1)と同様の手順でメタノールによる抽出を行い、最終的に菌体またはクリームの乾燥重量1mgあたり1mlの量になるようにそれぞれの抽出液の濃度を調整した。濃度調整した各抽出液100μlを供試し、それらの紫外線吸収スペクトルおよび320-400nmおよび340-380nmの波長域のピーク面積を(5)と同じ分光光度計及び解析ソフトで測定・比較した(図7)。
ここで、各サンプルについて、320-400nmのピーク面積、340-380nmのピーク面積、及び、[340-380nmのピーク面積]/[320-400nmのピーク面積]の比を以下に示す。
(6) Difference in absorption spectrum between commercially available cosmetics and ultraviolet absorbent composition of the present invention Dry cells of Methylobacterium sp. W-213 obtained by the same method as in (1) above and commercially available sunscreen creams A and B Extracted with methanol in the same procedure as in (1), and adjusted the concentration of each extract to 1 ml per 1 mg of dry weight of cells or cream. . 100 μl of each concentration-adjusted extract was tested, and the ultraviolet absorption spectrum and peak areas in the wavelength range of 320-400 nm and 340-380 nm were measured and compared using the same spectrophotometer and analysis software as in (5) (FIG. 7). ).
Here, for each sample, the peak area of 320-400 nm, the peak area of 340-380 nm, and the ratio of [peak area of 340-380 nm] / [peak area of 320-400 nm] are shown below.
上記の通り、本発明成分はUV−Aの領域に特異的吸収能を持つため、例えば、UV−Aの領域に吸収能を持たない廉価品の市販日焼け止め製品に添加すること等により、天然由来成分として優れた効果を発揮することができる。 As described above, since the component of the present invention has a specific absorption ability in the UV-A region, for example, by adding it to an inexpensive commercial sunscreen product having no absorption ability in the UV-A region, An excellent effect can be exhibited as a derived component.
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WO2017051875A1 (en) * | 2015-09-24 | 2017-03-30 | 昭和電工株式会社 | Compound exhibiting ultraviolet absorption ability or salt thereof, production method therefor, external preparation for skin, and cosmetic |
WO2017170713A1 (en) * | 2016-03-31 | 2017-10-05 | 国立研究開発法人農業・食品産業技術総合研究機構 | Anti-photoaging agent |
WO2017170722A1 (en) * | 2016-03-31 | 2017-10-05 | 国立研究開発法人農業・食品産業技術総合研究機構 | Anti-inflammatory agent |
JP2019043859A (en) * | 2017-08-30 | 2019-03-22 | 株式会社 ナチュラル | Ultraviolet-blocking cosmetic composition |
FR3143982A1 (en) | 2022-12-22 | 2024-06-28 | Dic Corporation | SUN PROTECTION AGENT |
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WO2017051875A1 (en) * | 2015-09-24 | 2017-03-30 | 昭和電工株式会社 | Compound exhibiting ultraviolet absorption ability or salt thereof, production method therefor, external preparation for skin, and cosmetic |
JPWO2017051875A1 (en) * | 2015-09-24 | 2018-07-26 | 国立研究開発法人農業・食品産業技術総合研究機構 | UV-absorbing compound or salt thereof, method for producing the same, external preparation for skin, cosmetic |
WO2017170713A1 (en) * | 2016-03-31 | 2017-10-05 | 国立研究開発法人農業・食品産業技術総合研究機構 | Anti-photoaging agent |
WO2017170722A1 (en) * | 2016-03-31 | 2017-10-05 | 国立研究開発法人農業・食品産業技術総合研究機構 | Anti-inflammatory agent |
JP2017186258A (en) * | 2016-03-31 | 2017-10-12 | 昭和電工株式会社 | Anti-photoaging agent |
JP2017186259A (en) * | 2016-03-31 | 2017-10-12 | 昭和電工株式会社 | Antiinflammatory agent |
JP2019043859A (en) * | 2017-08-30 | 2019-03-22 | 株式会社 ナチュラル | Ultraviolet-blocking cosmetic composition |
FR3143982A1 (en) | 2022-12-22 | 2024-06-28 | Dic Corporation | SUN PROTECTION AGENT |
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