JP6288686B2 - Egg shell membrane protein solubilized product and method for producing the same, composition containing the eggshell membrane protein solubilized product, composition for hair cosmetics, and hair treatment method using the composition for hair cosmetics - Google Patents
Egg shell membrane protein solubilized product and method for producing the same, composition containing the eggshell membrane protein solubilized product, composition for hair cosmetics, and hair treatment method using the composition for hair cosmetics Download PDFInfo
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- 229940012466 egg shell membrane Drugs 0.000 title claims description 202
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Description
本発明は、タンパク質を水溶化した卵殻膜タンパク質可溶化物及びその製造方法に関する。また、該卵殻膜タンパク質可溶化物を含有する組成物、毛髪化粧料用組成物及び該毛髪化粧料用組成物を用いた毛髪の処理方法に関する。 The present invention relates to a solubilized egg shell membrane protein obtained by water-solubilizing proteins and a method for producing the same. In addition, the present invention relates to a composition containing the solubilized eggshell membrane protein, a composition for hair cosmetics, and a hair treatment method using the composition for hair cosmetics.
卵殻膜に含まれるタンパク質成分やペプチド成分は、線維芽細胞増殖作用、保湿作用、活性酵素除去作用、細胞賦活作用等を有することが知られており、従来、化粧品原料や医薬品原料、食糧品原料等として広く利用されている。特に、卵殻膜は、システイン及びシスチンの含有量が高く、アミノ酸組成が毛髪と似ているという性質を有するために、毛髪化粧料の原料として利用されている。
例えば、特許文献１や２には、卵殻膜に含まれるタンパク質成分やペプチド成分を配合したパーマネントウェーブ用剤や毛髪処理剤が開示されている。
Protein components and peptide components contained in eggshell membranes are known to have fibroblast proliferation action, moisturizing action, active enzyme removal action, cell activation action, etc., and conventionally, cosmetic raw materials, pharmaceutical raw materials, food raw materials Etc. are widely used. In particular, eggshell membranes are used as a raw material for hair cosmetics because of their high cysteine and cystine contents and amino acid composition similar to that of hair.
For example, Patent Documents 1 and 2 disclose a permanent wave agent and a hair treatment agent in which protein components and peptide components contained in eggshell membranes are blended.
一方、化粧品や毛髪化粧料、医薬品、食糧品等の応用製品に配合される卵殻膜は、原料として配合しやすくするために、一般的に卵殻膜に含まれるタンパク質のペプチド結合を加水分解して、より分子量の小さい可溶性の卵殻膜タンパク質可溶化物として用いられている。 On the other hand, eggshell membranes blended in applied products such as cosmetics, hair cosmetics, pharmaceuticals, and foodstuffs generally hydrolyze peptide bonds of proteins contained in eggshell membranes in order to facilitate blending as raw materials. It is used as a soluble eggshell membrane protein lysate with a lower molecular weight.
タンパク質の加水分解方法として、様々な加水分解処理法等が知られている。
しかしながら、酸を用いて加水分解する方法では、装置が腐食しやすく、また卵殻膜のタンパク質が過剰に分解し、目的とする分子量分布を外れて低分子化しすぎたり、褐色に着色したり、目的とする加水分解物以外にも副生成物が生成しやすいという問題があった。
また、卵殻膜をタンパク質分解酵素で処理する酵素加水分解方法として、例えば、特許文献３には卵殻膜懸濁液中の卵殻膜に対してタンパク質分解酵素を１〜１０重量％の割合で加えてｐＨを６〜１２に調整し、温度３０〜６０℃にして酵素処理を６〜２４時間行い、乾燥させて、卵殻膜を原料とする可溶化卵殻膜の製法が報告されている。しかしながら、分子量の大きな卵殻膜タンパク質の可溶化物は得られにくく、触媒であるタンパク質分解酵素自体が高価であることから、製造コストが高くなるという問題があった。
Various methods for hydrolyzing proteins are known.
However, in the method of hydrolyzing with an acid, the device is easily corroded, the protein of eggshell membrane is excessively decomposed, deviating from the target molecular weight distribution, becoming too low in molecular weight, colored in brown, In addition to the hydrolyzate, there is a problem that by-products are easily generated.
In addition, as an enzyme hydrolysis method for treating an eggshell membrane with a proteolytic enzyme, for example, Patent Document 3 includes adding a proteolytic enzyme at a ratio of 1 to 10% by weight to the eggshell membrane in the eggshell membrane suspension. A method for producing a solubilized eggshell membrane using an eggshell membrane as a raw material has been reported, wherein the pH is adjusted to 6 to 12, the enzyme treatment is carried out at a temperature of 30 to 60 ° C. for 6 to 24 hours, and dried. However, a solubilized product of eggshell membrane protein having a large molecular weight is difficult to obtain, and the proteolytic enzyme itself, which is a catalyst, is expensive.
一方、卵殻膜タンパク質を、アルカリを用いて加水分解する方法も知られている。卵殻膜タンパク質の加水分解方法として、例えば、特許文献４では溶媒としてアルカリ性含水有機溶媒を使用して加水分解を行い、得られた分解液を陰イオン交換樹脂と接触させる可溶化卵殻膜の製法が報告されている。 On the other hand, a method of hydrolyzing eggshell membrane protein using an alkali is also known. As a hydrolysis method of eggshell membrane protein, for example, Patent Document 4 discloses a method for producing a solubilized eggshell membrane in which hydrolysis is performed using an alkaline hydrous organic solvent as a solvent, and the resulting decomposition solution is brought into contact with an anion exchange resin. It has been reported.
しかしながら、従来のアルカリ加水分解法では、卵殻膜タンパク質を、室温〜１００℃程度で長時間処理し、低分子量化させないと、可溶化させることができなかった。そのため、例えば、ダメージの大きい毛髪に対しては、十分な補修、保護効果を付与することができなかった。また、卵殻膜タンパク質加水分解物の着色を抑制するために有機溶媒を使用したりするため、これらの処理に余剰コストがかかり、製造コストも高くなりやすいという問題があった。 However, in the conventional alkaline hydrolysis method, eggshell membrane proteins could not be solubilized unless they were treated at room temperature to about 100 ° C. for a long time to reduce the molecular weight. Therefore, for example, sufficient repair and protection effects could not be imparted to hair with great damage. In addition, since an organic solvent is used to suppress coloring of the eggshell membrane protein hydrolyzate, there is a problem in that these treatments require extra costs and manufacturing costs tend to be high.
かかる状況下、本発明の目的は、従来の卵殻膜タンパク質の可溶化物と比較して、分子量が大きい卵殻膜タンパク質可溶化物及びその製造方法、並びに、該卵殻膜タンパク質可溶化物を含有する組成物を提供することである。 Under such circumstances, an object of the present invention is to contain an egg shell membrane protein lysate having a large molecular weight, a method for producing the same, and the eggshell membrane protein lysate, as compared with a conventional lysate of eggshell membrane protein. It is to provide a composition.
本発明者は、上記課題を解決すべく鋭意研究を重ねた結果、アルカリ条件下においても、急速加熱することにより、低分子量化させず、かつシステイン及びシスチンを残存させたまま、卵殻膜タンパク質を可溶化できることを見出し、下記の発明が上記目的に合致することを見出し、本発明に至った。 As a result of intensive research to solve the above-mentioned problems, the present inventor has found that eggshell membrane proteins can be obtained by rapid heating even under alkaline conditions without reducing the molecular weight and leaving cysteine and cystine remaining. The inventors have found that it can be solubilized, and found that the following inventions meet the above-mentioned purpose, and have reached the present invention.
すなわち、本発明は、以下の発明に係るものである。
＜１＞ 下記条件にて測定されたサイズ排除クロマトグラムにおいて、保持時間１６分間より短い時間にピークＡを有する卵殻膜タンパク質可溶化物。
（測定条件）
測定方法：サイズ排除クロマトグラフィー
カラム：ＧＥ製、商品名「Ｓｕｐｅｒｄｅｘ ｐｅｐｔｉｄｅ ＨＲ １０／３００」１本
溶離液：３０％アセトニトリル―７０％水−０．１％トリフルオロ酢酸
流量：０．５ｍｌ／ｍｉｎ
カラム温度：４０℃
検出器：紫外吸光光度計（測定波長２１４ｎｍ）、
試料濃度：５〜１０ｍｇ／ｍｌ
試料注入量：２５μＬ
＜２＞ 前記条件にて測定されたサイズ排除クロマトグラムにおいて、
総ピーク面積に対する、前記ピークＡの面積の比が９％以上である前記＜１＞に記載の卵殻膜タンパク質可溶化物。
＜３＞ 卵殻膜タンパク質可溶化物の重量平均分子量は、９０００以上１４０００以下である前記＜１＞又は＜２＞に記載の卵殻膜タンパク質可溶化物。
＜４＞ 原料に対し、システイン及びシスチンの含有量の残存率が３０％以上である前記＜１＞から＜３＞のいずれかに記載の卵殻膜タンパク質可溶化物。
＜５＞ 前記＜１＞から＜４＞のいずれかに記載の卵殻膜タンパク質可溶化物を含有する組成物。
＜６＞ 前記＜１＞から＜４＞のいずれかに記載の卵殻膜タンパク質可溶化物を含有する毛髪化粧料用組成物。
＜７＞ 前記＜６＞に記載の毛髪化粧料用組成物を用いた毛髪の処理方法。
＜８＞ アルカリ条件に調整した卵殻膜タンパク質の懸濁液を処理温度１３０℃以上１６０℃以下で、かつ、当該処理温度に５分以内に到達するよう加熱することによって卵殻膜タンパク質を可溶化する工程を有する卵殻膜タンパク質可溶化物の製造方法。
＜９＞ 前記卵殻膜タンパク質の懸濁液の加熱方法が、加圧した水蒸気と前記卵殻膜タンパク質の懸濁液との接触による加熱である前記＜８＞に記載の卵殻膜タンパク質可溶化物の製造方法。
＜１０＞ 原料に対し、システイン及びシスチンの含有量の残存量が３０％以上となるよう卵殻膜タンパク質を可溶化する前記＜８＞又は＜９＞に記載の卵殻膜タンパク質可溶化物の製造方法。
That is, the present invention relates to the following inventions.
<1> An eggshell membrane protein lysate having a peak A at a retention time shorter than 16 minutes in a size exclusion chromatogram measured under the following conditions.
(Measurement condition)
Measurement method: Size exclusion chromatography column: GE, trade name “Superdex peptide HR 10/300” 1 eluent: 30% acetonitrile-70% water-0.1% trifluoroacetic acid flow rate: 0.5 ml / min
Column temperature: 40 ° C
Detector: ultraviolet absorptiometer (measurement wavelength 214 nm),
Sample concentration: 5-10 mg / ml
Sample injection volume: 25 μL
<2> In the size exclusion chromatogram measured under the above conditions,
The eggshell membrane protein lysate according to <1>, wherein the ratio of the area of the peak A to the total peak area is 9% or more.
<3> The eggshell membrane protein lysate according to <1> or <2>, wherein the eggshell membrane protein lysate has a weight average molecular weight of 9000 or more and 14,000 or less.
<4> The eggshell membrane protein solubilized product according to any one of <1> to <3>, wherein the residual ratio of the content of cysteine and cystine is 30% or more with respect to the raw material.
<5> A composition comprising the solubilized eggshell membrane protein according to any one of <1> to <4>.
<6> A hair cosmetic composition containing the eggshell membrane protein solubilized product according to any one of <1> to <4>.
<7> A hair treatment method using the hair cosmetic composition according to <6>.
<8> The eggshell membrane protein is solubilized by heating the suspension of eggshell membrane protein adjusted to an alkaline condition at a treatment temperature of 130 ° C. to 160 ° C. so as to reach the treatment temperature within 5 minutes. The manufacturing method of the eggshell membrane protein lysate which has a process.
<9> The eggshell membrane protein lysate according to <8>, wherein the heating method of the eggshell membrane protein suspension is heating by contact with pressurized water vapor and the eggshell membrane protein suspension. Production method.
<10> The method for producing a solubilized eggshell membrane protein according to <8> or <9>, wherein the eggshell membrane protein is solubilized so that the residual amount of cysteine and cystine is 30% or more with respect to the raw material .
本発明によれば、従来の卵殻膜タンパク質の可溶化物と比較して、分子量が大きい卵殻膜タンパク質可溶化物及びその製造方法が提供される。該卵殻膜タンパク質可溶化物は、例えば、化粧料用組成物や毛髪化粧料用組成物の原料として好適に使用できる。 According to the present invention, an eggshell membrane protein lysate having a large molecular weight and a method for producing the same are provided as compared with a conventional lysate of eggshell membrane protein. The eggshell membrane protein solubilized product can be suitably used, for example, as a raw material for a cosmetic composition or a hair cosmetic composition.
以下、本発明について例示物等を示して詳細に説明するが、本発明は以下の例示物等に限定されるものではなく、本発明の要旨を逸脱しない範囲において任意に変更して実施できる。なお、本明細書において、「〜」とはその前後の数値又は物理量を含む表現として用いるものとする。 Hereinafter, the present invention will be described in detail with reference to examples and the like, but the present invention is not limited to the following examples and the like, and can be arbitrarily modified and implemented without departing from the gist of the present invention. In the present specification, “to” is used as an expression including numerical values or physical quantities before and after.
［１］卵殻膜タンパク質可溶化物
本発明の卵殻膜タンパク質可溶化物（以下、「卵殻膜可溶化物」と称する場合がある。）は、下記条件にて測定されたサイズ排除クロマトグラムにおいて、保持時間１６分間より短い時間にピークＡを有することを特徴とする。
（測定条件）
測定方法：サイズ排除クロマトグラフィー
カラム：ＧＥ製、商品名「Ｓｕｐｅｒｄｅｘ ｐｅｐｔｉｄｅ ＨＲ １０／３００」１本
溶離液：３０％アセトニトリル―７０％水−０．１％トリフルオロ酢酸
流量：０．５ｍｌ／ｍｉｎ
カラム温度：４０℃
検出器：紫外吸光光度計（測定波長２１４ｎｍ）、
試料濃度：５〜１０ｍｇ／ｍｌ
試料注入量：２５μｌ
[1] Eggshell membrane protein solubilized product The eggshell membrane protein solubilized product of the present invention (hereinafter sometimes referred to as “eggshell membrane solubilized product”) is a size exclusion chromatogram measured under the following conditions: It has a peak A at a holding time shorter than 16 minutes.
(Measurement condition)
Measurement method: Size exclusion chromatography column: GE, trade name “Superdex peptide HR 10/300” 1 eluent: 30% acetonitrile-70% water-0.1% trifluoroacetic acid flow rate: 0.5 ml / min
Column temperature: 40 ° C
Detector: ultraviolet absorptiometer (measurement wavelength 214 nm),
Sample concentration: 5-10 mg / ml
Sample injection volume: 25 μl
本明細書において、「卵殻膜タンパク質」は卵殻膜に含まれるタンパク質成分を意味する。卵殻膜とは、卵殻膜は卵殻の内側にある繊維質の薄膜であり、外卵殻膜及び内卵殻膜の二層の網目状構造からなる。卵殻膜の主成分はタンパク質であるが、卵殻から卵殻膜を分離する際に卵殻由来のカルシウム成分を含むことがある。
また、「卵殻膜タンパク質可溶化物」とは、原料の卵殻膜タンパク質が水に可溶化できる状態に変化したものを意味する。ここで、本発明において、「水に可溶化できる状態」には、卵殻膜タンパク質由来の成分が、水に溶解する状態だけでなく、自然沈降せずに水に分散する状態を含む。
In the present specification, “eggshell membrane protein” means a protein component contained in eggshell membrane. The eggshell membrane is a fibrous thin film inside the eggshell, and has a two-layer network structure of an outer eggshell membrane and an inner eggshell membrane. The main component of the eggshell membrane is protein, but when the eggshell membrane is separated from the eggshell, it may contain a calcium component derived from eggshell.
The “eggshell membrane protein solubilized product” means a product in which the raw eggshell membrane protein is changed to a state in which it can be solubilized in water. Here, in the present invention, the “state that can be solubilized in water” includes not only the state in which the eggshell membrane protein-derived component is dissolved in water but also the state in which it is dispersed in water without spontaneous sedimentation.
図１に、上記条件にて測定された本発明の卵殻膜タンパク質可溶化物のサイズ排除クロマトグラムの一例を示す。図１の卵殻膜タンパク質可溶化物の詳細については、実施例１にて後述するが、図１の卵殻膜タンパク質可溶化物は、保持時間１５．０〜１５．２分の間にピークトップを持つピークＡを有する。
ここで、上記測定条件にて測定されたサイズ排除クロマトグラムにおいて、排除限界分子量は２００００であり、分子量２００００以上のものは、保持時間１６分より短い時間にピークＡとして検出される。ピークＡは、いわゆる排除限界分子量のピークである。
すなわち、本発明の卵殻膜タンパク質可溶化物の特徴である、保持時間１６分より短い時間にピークＡを有するということは、本発明の卵殻膜タンパク質可溶化物は、一般的に可溶化しにくい高分子量の成分を含んでいるにもかかわらず可溶化しているということである。
FIG. 1 shows an example of the size exclusion chromatogram of the solubilized eggshell membrane protein of the present invention measured under the above conditions. The details of the eggshell membrane protein lysate of FIG. 1 will be described later in Example 1. The eggshell membrane protein lysate of FIG. 1 has a peak top during the retention time of 15.0 to 15.2 minutes. It has a peak A.
Here, in the size exclusion chromatogram measured under the above measurement conditions, the exclusion limit molecular weight is 20000, and those having a molecular weight of 20000 or more are detected as peak A in a time shorter than the retention time of 16 minutes. Peak A is a so-called exclusion limit molecular weight peak.
That is, having a peak A in a time shorter than the retention time of 16 minutes, which is a characteristic of the eggshell membrane protein lysate of the present invention, the eggshell membrane protein lysate of the present invention is generally difficult to solubilize. It means that it is solubilized despite containing high molecular weight components.
一方、図４に、従来のアルカリ加水分解で製造された卵殻膜タンパク質の可溶化物のサイズ排除クロマトグラムの一例を示す。図４の卵殻膜タンパク質可溶化物の詳細については、比較例１にて後述するが、図４の卵殻膜タンパク質可溶化物は、保持時間１６分より短い時間にピークＡは検出されず、全体的に低分子量化することで可溶化していることがわかる。 On the other hand, FIG. 4 shows an example of a size exclusion chromatogram of a solubilized product of eggshell membrane protein produced by conventional alkaline hydrolysis. The details of the solubilized eggshell membrane protein of FIG. 4 will be described later in Comparative Example 1. However, the solubilized eggshell membrane protein of FIG. It can be seen that solubilization is achieved by lowering the molecular weight.
このように本発明の卵殻膜タンパク質可溶化物は、従来の卵殻膜タンパク質可溶化物と比較して高分子量成分を含むにもかかわらず、水に可溶化することができ、従来の卵殻膜タンパク質可溶化物とは異なる性質を有したものである。 Thus, the eggshell membrane protein solubilized product of the present invention can be solubilized in water despite containing a high molecular weight component as compared with the conventional eggshell membrane protein solubilized product. It has different properties from the solubilized product.
なお、サイズ排除クロマトグラフィーは、例えば、株式会社島津製作所社製高速液体クロマトグラフ、装置名「プロミネンス」等により測定することができる。 The size exclusion chromatography can be measured by, for example, a high performance liquid chromatograph manufactured by Shimadzu Corporation, an apparatus name “Prominence” and the like.
化粧料用組成物や毛髪化粧料用組成物の原料としては、高分子量成分をより含有することが好ましく、上記条件にて測定されたサイズ排除クロマトグラムにおいて、総ピーク面積に対する、ピークＡの面積の比（（ピークＡの面積／総ピーク面積）×１００）が３％以上であることが好ましく、５％以上であることがより好ましく、９％以上であることがさらに好ましい。
高分子量成分をより含有することで、爪、まつげ、まゆげ、毛髪等に対してより高い保護効果が期待できる。
As a raw material for a cosmetic composition or a hair cosmetic composition, it is preferable to further contain a high molecular weight component, and in the size exclusion chromatogram measured under the above conditions, the area of peak A relative to the total peak area Ratio ((Area of peak A / total peak area) × 100) is preferably 3% or more, more preferably 5% or more, and further preferably 9% or more.
By containing a higher molecular weight component, a higher protective effect can be expected for nails, eyelashes, eyebrows, hair and the like.
なお、「総ピーク面積」とは、測定開始から測定終了までの全てのピークの面積の和を意味する。測定終了点は、その保持時間以降のピーク強度及びピークの傾きが実質的に０となる点とする。
また、「ピークＡの面積」は、ピークＡの立ち上がりの点（測定開始後、ピーク強度が実質的に０より大きくなる最初の点）の保持時間から、ピークＡのピークトップ検出後、最初にピークの傾きが０となる点（変曲点）の保持時間までの間の面積とする。
“Total peak area” means the sum of the areas of all peaks from the start of measurement to the end of measurement. The measurement end point is a point at which the peak intensity and the slope of the peak after the holding time become substantially zero.
“Area of peak A” is determined from the retention time at the rising point of peak A (the first point where the peak intensity is substantially greater than 0 after the start of measurement), after the peak top of peak A is detected. The area until the holding time of the point (inflection point) at which the peak slope becomes 0 is taken.
また、本発明の卵殻膜可溶化物は、上記の条件において、サイズ排除クロマトグラムにおいて、保持時間１６分より短い時間にピークＡを有すれば、使用目的等に応じて、重量平均分子量は適宜調整することができ、卵殻膜可溶化物の重量平均分子量の上限及び下限は、使用目的に応じて、安全面等を考慮して適宜決定すればよい。例えば、重量平均分子量の上限は、感作性の発現しない範囲で、適宜決定すればよい。
化粧料用途や毛髪化粧料用途においては、保護効果をより高めるために、卵殻膜可溶化物の重量平均分子量が９０００〜１４０００であることが好ましい。なお、本明細書において、卵殻膜タンパク質可溶化物の重量平均分子量とは、上記の条件にて測定されたサイズ排除クロマトグラムより算出した重量平均分子量のことである。
In addition, the solubilized eggshell membranes of the present invention has a weight average molecular weight as appropriate depending on the purpose of use, etc., as long as it has a peak A in a size exclusion chromatogram with a retention time shorter than 16 minutes. The upper limit and lower limit of the weight average molecular weight of the solubilized eggshell membranes can be appropriately determined in consideration of safety and the like according to the purpose of use. For example, the upper limit of the weight average molecular weight may be appropriately determined within a range where sensitization does not occur.
In cosmetic applications and hair cosmetic applications, the eggshell membrane solubilized product preferably has a weight average molecular weight of 9000 to 14000 in order to further enhance the protective effect. In the present specification, the weight average molecular weight of the solubilized eggshell membrane protein refers to the weight average molecular weight calculated from the size exclusion chromatogram measured under the above conditions.
また、化粧料用途や毛髪化粧用組成物の原料として用いた場合、保護効果に加えて補修効果を向上させるためには、卵殻膜タンパク質の可溶化物は、システイン及びシスチンを多く含有することが好ましい。本発明の卵殻膜可溶化物は、高分子量成分を多く含むため、システイン及びシスチンの含有量を高めやすい。システイン及びシスチンの含有量を高めることで、爪、まつげ、まゆげ、毛髪等に対してより高い保護効果や補修効果が期待できる。 In addition, when used as a raw material for cosmetic applications and hair cosmetic compositions, solubilized eggshell membrane proteins may contain a large amount of cysteine and cystine in order to improve the repair effect in addition to the protective effect. preferable. Since the solubilized eggshell membranes of the present invention contains a large amount of high molecular weight components, it is easy to increase the content of cysteine and cystine. By increasing the content of cysteine and cystine, higher protection and repair effects can be expected for nails, eyelashes, eyebrows, hair, and the like.
本発明の卵殻膜可溶化物のシステイン及びシスチンの含有量は、原料に対しシステイン及びシスチンの含有量の残存率が３０％以上であることが好ましく、４０％以上であることがより好ましい。システイン及びシスチンの含有量の算出方法、残存率については、実施例にて後述する。 The content of cysteine and cystine in the solubilized eggshell membranes of the present invention is preferably such that the residual ratio of the content of cysteine and cystine is 30% or more, more preferably 40% or more. The calculation method of the content of cysteine and cystine and the residual rate will be described later in Examples.
また、卵殻膜可溶化物のシステイン及びシスチンの含有量が１．５質量％以上であることが好ましく、より好ましくは２．０質量％以上であり、４．０質量％がさらに好ましい。 Further, the content of cysteine and cystine in the solubilized eggshell membrane is preferably 1.5% by mass or more, more preferably 2.0% by mass or more, and further preferably 4.0% by mass.
本発明の卵殻膜可溶化物は、液状（可溶化した状態）で用いてもよいし、乾燥させて粉末や顆粒などの固形物として使用してもよい。
本発明の卵殻膜可溶化物は、水に可溶であり、着色がほとんどないと共に、人体に対する毒性や刺激性が少ないため、各種化粧料用組成物、医薬品用組成物（外用剤、服用剤）、機能性食品等の用途に使用することができる。詳しくは、［３］本発明の卵殻膜タンパク質可溶化物を含む組成物において、後述する。
The eggshell membrane solubilized product of the present invention may be used in a liquid state (solubilized state) or may be dried and used as a solid material such as powder or granule.
The eggshell membrane solubilized product of the present invention is soluble in water, hardly colored, and has little toxicity and irritation to the human body. Therefore, various cosmetic compositions and pharmaceutical compositions (external preparations, medications) ), Can be used for functional foods. Specifically, in [3] the composition containing the solubilized eggshell membrane protein of the present invention, it will be described later.
また、本発明の卵殻膜可溶化物の製造方法は、後述する本発明の卵殻膜タンパク質可溶化物の製造方法で好適に製造することができる。 Moreover, the manufacturing method of the eggshell membrane solubilized product of this invention can be suitably manufactured with the manufacturing method of the eggshell membrane protein solubilized product of this invention mentioned later.
［２］卵殻膜タンパク質可溶化物の製造方法
本発明の卵殻膜タンパク質可溶化物の製造方法（以下、「本発明の製造方法」と称する場合がある。）は、アルカリ条件に調整した卵殻膜タンパク質懸濁液（以下、単に「懸濁液」と称する場合がある。）を温度１３０℃以上１６０℃以下で、かつ、当該処理温度に５分以内に到達するように加熱することによって卵殻膜タンパク質を可溶化する工程を有することを特徴とする。
[2] Method for Producing Eggshell Membrane Protein Solubilized Product The method for producing the solubilized eggshell membrane protein product of the present invention (hereinafter sometimes referred to as “the production method of the present invention”) is an eggshell membrane adjusted to an alkaline condition. By heating a protein suspension (hereinafter sometimes simply referred to as “suspension”) at a temperature of 130 ° C. or higher and 160 ° C. or lower and reaching the treatment temperature within 5 minutes, the eggshell membrane It has the process of solubilizing protein.
本発明の製造方法により、上述した本発明の卵殻膜タンパク質の可溶化物を好適に製造できる。 By the production method of the present invention, the above-described solubilized eggshell membrane protein of the present invention can be suitably produced.
本発明の製造方法の特徴のひとつは、１３０℃〜１６０℃の設定温度に急速加熱し、卵殻膜タンパク質の可溶化をすることである。１３０℃〜１６０℃の設定温度に急速加熱し、反応系内を急速に高温することで、変性や凝集を抑制しつつ、タンパク質の構造を変化に伴う可溶化反応を優先的に進行させつつ、適度な加水分解をもされるので、短時間であっても、十分に可溶化させることができると推察される。また、アルカリ条件下で急速加熱し、短時間で卵殻膜タンパク質を可溶化することで、タンパク質の変性を抑制しやすく、タンパク質可溶化物同士が電荷によって反発しあうため凝集を抑制しやすい傾向にあるため、従来のアルカリの処理液を室温ないしは８０〜１００℃程度の温度で、長時間、処理することによって得られる卵殻膜タンパク質加水分解物と比較して、分子量の大きなものであっても可溶化できる。 One of the features of the production method of the present invention is that the eggshell membrane protein is solubilized by rapid heating to a set temperature of 130 ° C to 160 ° C. While rapidly heating to a set temperature of 130 ° C. to 160 ° C. and rapidly raising the temperature in the reaction system, while suppressing denaturation and aggregation, while preferentially proceeding solubilization reaction accompanying changes in protein structure, Since it is moderately hydrolyzed, it is presumed that it can be sufficiently solubilized even in a short time. In addition, rapid heating under alkaline conditions and solubilization of eggshell membrane protein in a short time makes it easy to suppress protein denaturation and tends to suppress aggregation because protein lysates repel each other due to charge. Therefore, it may have a higher molecular weight than the eggshell membrane protein hydrolyzate obtained by treating a conventional alkali treatment solution at room temperature or at a temperature of about 80 to 100 ° C. for a long time. Can be solubilized.
また、酸条件下での可溶化方法と比較して、分子量分布の狭いものが得られやすいため、高分子量化成分を多く含む卵殻膜可溶化物が得られやすく、副生成物の生成も抑制することができる。 Compared with the solubilization method under acid conditions, it is easy to obtain a narrow molecular weight distribution, so it is easy to obtain solubilized eggshell membranes containing a large amount of high molecular weight components, and also suppresses the formation of by-products. can do.
また、従来の製法で得られる卵殻膜可溶化物は、可溶化物を得るためにアルカリ条件下で長時間の加熱処理が必要となるために、システイン及びシスチンが分解し、システイン及びシスチンの含有量が低下しやすかった。
本発明の製造方法では、短時間で卵殻膜タンパク質を可溶化させることができるので、従来のアルカリ加水分解法では分解されやすい、卵殻膜の機能性に影響するシスチン及びシステインも分解されにくい。
In addition, the eggshell membrane solubilizate obtained by the conventional method requires heat treatment under alkaline conditions for a long time in order to obtain a solubilizate, so that cysteine and cystine are decomposed and cysteine and cystine are contained. The amount was easy to fall.
Since the eggshell membrane protein can be solubilized in a short time in the production method of the present invention, cystine and cysteine that affect the functionality of the eggshell membrane, which are easily degraded by the conventional alkaline hydrolysis method, are also hardly degraded.
本発明の製造方法で使用される懸濁液はアルカリ条件に調整される。本発明の卵殻膜可溶化物をより効率に得るためには、アルカリ条件として、ｐＨは８．０以上が好ましく、ｐＨ９．０以上がより好ましく、ｐＨ１０．０以上がさらに好ましく、ｐＨ１１．０以上が最も好ましい。なお、本発明におけるｐＨは、卵殻膜タンパク質懸濁液をｐＨメータで測定した値である。 The suspension used in the production method of the present invention is adjusted to alkaline conditions. In order to obtain the eggshell membrane lysate of the present invention more efficiently, the pH is preferably 8.0 or higher, more preferably pH 9.0 or higher, further preferably pH 10.0 or higher, and pH 11.0 or higher as alkaline conditions. Is most preferred. The pH in the present invention is a value obtained by measuring an eggshell membrane protein suspension with a pH meter.
処理温度は、使用目的に応じて１３０℃〜１６０℃の間で適宜選択され、処理温度への到達時間は、５分以内である。処理温度が温度１３０℃未満の場合には可溶化反応が徐々に起こり、反応に時間を有し、低分子量化しやすい。また、１６０℃を超えると、可溶化反応より卵殻膜タンパク質の変性及び過度の加水分解が優先的に起こりやすく、本発明の卵殻膜タンパク質を得にくい。
また、目的とする温度までに到達する時間が長すぎると、可溶化反応が徐々に起こるために、卵殻膜タンパク質の変性及び過度の加水分解が起こりやすくなり、本発明の卵殻膜タンパク質を得るのが困難である。
The treatment temperature is appropriately selected between 130 ° C. and 160 ° C. according to the purpose of use, and the time to reach the treatment temperature is within 5 minutes. When the treatment temperature is less than 130 ° C., the solubilization reaction occurs gradually, the reaction takes time, and the molecular weight tends to be lowered. Moreover, when it exceeds 160 degreeC, the modification | denaturation and excessive hydrolysis of eggshell membrane protein will occur preferentially over solubilization reaction, and it will be difficult to obtain the eggshell membrane protein of this invention.
Further, if the time to reach the target temperature is too long, the solubilization reaction occurs gradually, so that the eggshell membrane protein is easily denatured and excessively hydrolyzed, and the eggshell membrane protein of the present invention is obtained. Is difficult.
ここで、卵殻膜タンパク質の可溶化の程度は、懸濁液のｐＨと処理温度に依存して、好適な処理温度範囲が異なる。例えば、ｐＨ８．０以上１２以下の範囲における好適な条件は、処理温度が１５０〜１６０℃である。また、ｐＨ１２以上範囲における好適な条件は、処理温度が１３０〜１５０℃である。 Here, the degree of solubilization of the eggshell membrane protein varies depending on the pH of the suspension and the treatment temperature, and the preferred treatment temperature range is different. For example, as for the suitable conditions in the range of pH 8.0 or more and 12 or less, process temperature is 150-160 degreeC. Moreover, as for the suitable conditions in pH12 or more range, process temperature is 130-150 degreeC.
また、本発明の卵殻膜可溶化物をより効率に得るためには、処理温度への到達時間は１分以内が好適である。 Moreover, in order to obtain the eggshell membrane solubilizate of the present invention more efficiently, the time to reach the treatment temperature is preferably within 1 minute.
なお、「処理温度」とは、急速加熱のときの最高到達温度のことであり、装置により試料が加熱される箇所における試料の測定温度である。本発明においては、処理温度は、１３０℃〜１６０℃の間から適宜決定される。また、昇温の速度は、処理温度への到達時間は５分以内となるように、目的等を考慮して適宜調節可能である。 The “treatment temperature” refers to the highest temperature reached at the time of rapid heating, and is the measurement temperature of the sample at the location where the sample is heated by the apparatus. In the present invention, the treatment temperature is appropriately determined from 130 ° C to 160 ° C. In addition, the rate of temperature rise can be appropriately adjusted in consideration of the purpose and the like so that the time to reach the processing temperature is within 5 minutes.
また、処理温度に到達後、速やかに１００℃以下に冷却されることが好ましい。冷却は、熱交換器や減圧装置の装置を利用してもよい。 Moreover, it is preferable that it is cooled to 100 ° C. or less immediately after reaching the treatment temperature. For the cooling, a heat exchanger or a decompression device may be used.
また、本発明の目的を損なわない範囲で、１３０〜１６０℃の処理温度へ到達後、処理温度で保持してもよい。なお、「保持時間」には、処理時間到達後の処理温度で保持する時間のことであり、昇温時間は含まない。 Moreover, you may hold | maintain at processing temperature after reaching | attaining 130-160 degreeC processing temperature in the range which does not impair the objective of this invention. The “holding time” is the time for holding at the processing temperature after the processing time is reached, and does not include the temperature raising time.
加熱手段は上記処理条件に懸濁液を加熱できれば特に制限されず、加圧した水蒸気と前記卵殻膜タンパク質の懸濁液との接触による加熱や電磁誘導による加熱、マイクロウェーブ照射により加熱、熱交換機を介した間接的な加熱等が挙げられる。
このような加熱方法には、例えば、超高温瞬間殺菌装置（ＵＨＴ殺菌装置）を用いたり、マイクロ波加熱装置、誘導加熱装置等を用いることができる。なお、上述の処理条件を満たし、急速に卵殻膜タンパク質の懸濁液を加熱できるものであれば、例示した装置に限定されず、使用する設備に特に限定はなく、従来公知の装置を使用することができ、卵殻膜可溶化物の生産量を考慮して適宜好適な装置を選択すればよい。
The heating means is not particularly limited as long as the suspension can be heated to the above processing conditions. Heating by contact with pressurized water vapor and the eggshell membrane protein suspension, heating by electromagnetic induction, heating by microwave irradiation, heat exchanger Indirect heating through the like.
For such a heating method, for example, an ultra-high temperature instantaneous sterilizer (UHT sterilizer), a microwave heater, an induction heater, or the like can be used. In addition, as long as the above-described processing conditions are satisfied and the eggshell membrane protein suspension can be rapidly heated, the apparatus is not limited to the exemplified apparatus, and the equipment to be used is not particularly limited, and a conventionally known apparatus is used. In view of the production amount of the solubilized eggshell membranes, a suitable apparatus may be selected as appropriate.
この中でも、加熱手段は、加圧した水蒸気と前記卵殻膜タンパク質の懸濁液との接触による加熱であると、短時間で効率的に熱が伝わり、可溶化できるので、分子量の大きな卵殻膜可溶化物を得ることができ好ましい。また、多量の懸濁液を連続的に可溶化することができる点でも、加圧した水蒸気と前記卵殻膜タンパク質の懸濁液との接触による加熱が好適である。
加圧した水蒸気と前記卵殻膜タンパク質の懸濁液との接触による加熱の場合、加圧した水蒸気を懸濁液に吹き込んで加圧した水蒸気を懸濁液に接触させても、懸濁液を加圧した水蒸気中に吹き込んで加圧した水蒸気を懸濁液に接触させてもよい。
Among these, when the heating means is heating by contact with pressurized water vapor and the eggshell membrane protein suspension, heat can be efficiently transferred and solubilized in a short time, so that eggshell membranes having a large molecular weight can be used. A solvate can be obtained, which is preferable. In addition, heating by contact between pressurized water vapor and the eggshell membrane protein suspension is also preferable in that a large amount of suspension can be continuously solubilized.
In the case of heating by contact of pressurized water vapor with the eggshell membrane protein suspension, even if pressurized water vapor is blown into the suspension and the pressurized water vapor is brought into contact with the suspension, the suspension The pressurized water vapor blown into the pressurized water vapor may be brought into contact with the suspension.
加圧した水蒸気と前記卵殻膜タンパク質の懸濁液との接触による加熱ができる装置としては、例えば、超高温瞬間殺菌装置（ＵＨＴ殺菌装置）を用いることができる。
一般的に、超高温瞬間殺菌装置（ＵＨＴ殺菌装置）は、液状食品の殺菌に用いられ、液状食品に蒸気を吹き込む又は上記に液状食品を吹き込むことにより、食品の温度を極めて短時間で殺菌温度（１５０℃前後）まで加熱し、数秒保持することで、熱による食品のダメージを抑えつつ殺菌することができる装置である。本発明の製造方法は、一般的に殺菌のために用いられる超高温瞬間殺菌装置（ＵＨＴ殺菌装置）を転用し、卵殻膜可溶化物の製造のために用いることができる。
また、超高温瞬間殺菌装置の方式は特に限定されず、例えば、スチームインジェクション式でもよく、スチームインフュージョン式でもよい。このような装置として、例えば、イズミフードマシナリー製スチームインジェクションＳＤＩ―１５０殺菌テスト機等が挙げられる。
For example, an ultra-high temperature instantaneous sterilization apparatus (UHT sterilization apparatus) can be used as an apparatus capable of heating by contact with pressurized water vapor and the eggshell membrane protein suspension.
In general, an ultra-high temperature instant sterilizer (UHT sterilizer) is used for sterilization of liquid food, and the temperature of the food can be reduced to a sterilization temperature in a very short time by blowing steam into the liquid food or by blowing the liquid food above. It is an apparatus that can be sterilized while suppressing damage to food by heat by heating to (around 150 ° C.) and holding for several seconds. The production method of the present invention can be used for the production of solubilized eggshell membranes by diverting an ultra-high temperature instantaneous sterilization device (UHT sterilization device) generally used for sterilization.
Further, the method of the ultra-high temperature instantaneous sterilization apparatus is not particularly limited, and for example, a steam injection type or a steam infusion type may be used. Examples of such a device include a steam injection SDI-150 sterilization test machine manufactured by Izumi Food Machinery.
このような直接懸濁液に水蒸気を接触させて急速に温度を上げる装置は、懸濁液を連続的に処理できる点と、蒸気を直接接触させることにより懸濁液を効率よく撹拌、可溶化できる点で好適である。 Such a device that rapidly raises the temperature by bringing water vapor into contact with the suspension makes it possible to process the suspension continuously and efficiently agitates and solubilizes the suspension by bringing the vapor into direct contact. It is preferable in that it can be performed.
また、本発明の製造方法では、短時間で卵殻膜タンパク質を可溶化させることができるので、シスチン及びシステインが分解されにくいが、原料（可溶化処理前の卵殻膜タンパク質）に対し、システイン及びシスチンの含有量の残存量が３０％以上となるよう卵殻膜タンパク質を可溶化することが好ましい。このような製造方法は、化粧料用組成物や毛髪化粧料用組成物の原料となる卵殻膜可溶化物の製造に特に好適である。 Further, in the production method of the present invention, eggshell membrane proteins can be solubilized in a short time, so that cystine and cysteine are difficult to be decomposed. It is preferable to solubilize the eggshell membrane protein so that the residual amount of the content of is 30% or more. Such a production method is particularly suitable for the production of a solubilized eggshell membrane that is a raw material for a cosmetic composition or a hair cosmetic composition.
ｐＨ調整に使用するアルカリとしては、本発明の目的が達成される限り、水酸化ナトリウム、水酸化カリウム、水酸化カルシウム等の無機アルカリであっても、有機アルカリであってもよいが、通常、無機アルカリが用いられ、特に水酸化ナトリウムは好適なアルカリのひとつである。 The alkali used for pH adjustment may be an inorganic alkali such as sodium hydroxide, potassium hydroxide, calcium hydroxide, or an organic alkali as long as the object of the present invention is achieved. An inorganic alkali is used, and sodium hydroxide is one of the preferred alkalis.
ｐＨ調整に使用するアルカリの量は、懸濁液のｐＨが所望のアルカリ条件となるように調整され、原料となる未処理卵殻膜に含まれるカルシウム成分の量を考慮して適宜決定される。アルカリの濃度が高すぎると塩が残留しやすいため、アルカリ濃度３％以下の処理液を用いて卵殻膜タンパク質の懸濁液を調整することがより好ましい。 The amount of alkali used for pH adjustment is appropriately determined in consideration of the amount of calcium component contained in an untreated eggshell membrane as a raw material, so that the pH of the suspension is adjusted to a desired alkaline condition. If the alkali concentration is too high, the salt tends to remain, so it is more preferable to adjust the eggshell membrane protein suspension using a treatment solution having an alkali concentration of 3% or less.
また、本発明の卵殻膜可溶化物の製造方法は、上記工程を含めばよく、その前後に別の工程を設けてもよい。例えば、上記工程のあとに、脱塩処理等の精製を行ってもよい。 Moreover, the manufacturing method of the eggshell membrane solubilizate of this invention should just include the said process, and may provide another process before and after that. For example, purification such as desalting may be performed after the above step.
卵殻から卵殻膜を分離する際に卵殻由来のカルシウム成分を含むことがある。これをそのまま使用すると、カルシウム成分が残留し、得られる卵殻膜可溶化物の品質が劣化するおそれがある。そこで、このカルシウム成分を除去した卵殻膜タンパク質を使用することで卵殻膜可溶化物の品質が向上する。 When separating the eggshell membrane from the eggshell, it may contain calcium components derived from eggshell. If this is used as it is, the calcium component remains and the quality of the solubilized eggshell membranes may be deteriorated. Therefore, the quality of the solubilized eggshell membrane is improved by using the eggshell membrane protein from which the calcium component has been removed.
より高品質な卵殻膜可溶化物を得ることができる点で、原料の（未処理）卵殻膜タンパク質は、卵殻膜に含まれるカルシウム成分を酸性溶液で溶解させた後に、固液分離して得られる卵殻膜タンパク質であってもよい。すなわち、卵殻膜タンパク質を可溶化する工程の前工程として、原料である（未処理）卵殻膜タンパク質を酸性溶液で処理し、含有するカルシウム成分を溶解させ、固液分離することによりカルシウム成分を除去した卵殻膜タンパク質を得る工程を含んでもよい。 The raw (shelled) eggshell membrane protein is obtained by dissolving the calcium component contained in the eggshell membrane with an acidic solution and then separating it into solid and liquid in that a higher quality eggshell membrane solubilizate can be obtained. It may be an eggshell membrane protein. That is, as a pre-process of the step of solubilizing eggshell membrane protein, the raw (untreated) eggshell membrane protein is treated with an acidic solution, the calcium component contained is dissolved, and the calcium component is removed by solid-liquid separation. A step of obtaining a processed eggshell membrane protein may be included.
例えば、卵殻膜タンパク質を可溶化する工程の前工程として、原料である（未処理）卵殻膜タンパク質を乳酸で処理し、含有するカルシウム成分を溶解させ、固液分離することによりカルシウム成分を除去した卵殻膜タンパク質を得る工程を含んでもよい。
このようにすることで、カルシウム成分は乳酸カルシウムとして可溶化し、比重の低い不溶性のカルシウム塩は乳酸溶液に浮き、卵殻膜は乳酸溶液に沈降するために、容易に卵殻膜からカルシウム成分を除去することができる。
For example, as a pre-process of the process of solubilizing eggshell membrane protein, the raw (untreated) eggshell membrane protein is treated with lactic acid, the calcium component contained is dissolved, and the calcium component is removed by solid-liquid separation. You may include the process of obtaining eggshell membrane protein.
By doing so, the calcium component is solubilized as calcium lactate, the insoluble calcium salt with low specific gravity floats in the lactic acid solution, and the eggshell membrane settles in the lactic acid solution, so the calcium component is easily removed from the eggshell membrane. can do.
使用する酸としては、卵殻膜タンパク質の品質劣化を惹起するものでなく、カルシウム成分を溶解することができるものであればよく、塩酸等の無機酸であっても、乳酸やクエン酸、酢酸等の有機酸であってもよい。 The acid to be used does not cause deterioration in the quality of the eggshell membrane protein, and may be any acid that can dissolve the calcium component. Even if it is an inorganic acid such as hydrochloric acid, lactic acid, citric acid, acetic acid, etc. Organic acids may be used.
［３］本発明の卵殻膜タンパク質可溶化物を含む組成物
本発明の卵殻膜タンパク質可溶化物は、上述のように水溶性であり、着色がほとんどないと共に、人体に対する毒性や刺激性が少ない。そのため、本発明の卵殻膜可溶化物を含む組成物（以下、「本発明の組成物」と称する場合がある。）は、各種化粧料用組成物、医薬品用組成物（外用剤、服用剤）、機能性食品等の用途に使用することができ、その用途は特に限定されないが、好適な用途の一つとして、毛髪化粧料用組成物が挙げられる。
なお、本発明の卵殻膜可溶化物は、その用途に合わせて所望の分子量分布に調整して使用してもよい。
[3] Composition containing eggshell membrane protein solubilized product of the present invention The eggshell membrane protein solubilized product of the present invention is water-soluble as described above, hardly colored, and less toxic and irritating to the human body. . Therefore, the composition containing the solubilized eggshell membranes of the present invention (hereinafter sometimes referred to as “the composition of the present invention”) includes various cosmetic compositions and pharmaceutical compositions (external preparations, medications). ), And can be used for functional foods and the like, and the use thereof is not particularly limited, but one suitable application is a composition for hair cosmetics.
The solubilized eggshell membranes of the present invention may be used after adjusting to a desired molecular weight distribution according to the intended use.
また、本発明の組成物は、目的を損なわない範囲で、本発明の卵殻膜可溶化物のみならず、加水分解が進行した、より低分子化した成分を含んでいてもよい。 In addition, the composition of the present invention may contain not only the eggshell membrane solubilizate of the present invention but also a component having a lower molecular weight that has undergone hydrolysis as long as the object is not impaired.
（化粧料用組成物）
本発明の組成物は化粧料用組成物の用途に使用してもよい。本発明の卵殻膜可溶化物は、従来の卵殻膜可溶化物に比べて、分子量が大きいため、皮膚やまゆげ、まつげ、ネイル、毛髪の表面で膜を形成しやすく、保湿効果や保護効果を発揮しやすい。各種化粧料基材及び化粧料添加物に対する安定性も高いため、化粧料用組成物に好適に配合することができる。なお、本発明において、化粧料用組成物は、化粧品のみならず医薬部外品も含む。
(Cosmetic composition)
You may use the composition of this invention for the use of the composition for cosmetics. Since the solubilized eggshell membrane of the present invention has a higher molecular weight than conventional solubilized eggshell membranes, it is easy to form a film on the surface of the skin, eyelashes, eyelashes, nails and hair, and has a moisturizing effect and a protective effect. Easy to demonstrate. Since stability with respect to various cosmetic base materials and cosmetic additives is also high, it can be suitably blended in a cosmetic composition. In the present invention, the cosmetic composition includes not only cosmetics but also quasi drugs.
化粧料用組成物への卵殻膜可溶化物の配合割合は任意であるが、化粧料用組成物が、保湿作用等の卵殻膜可溶化物に由来する作用を発現する範囲で適宜選択される。卵殻膜可溶化物の化粧料用組成物に対する割合は、卵殻膜可溶化物における溶媒の割合や溶媒の種類等にもよるが、例えば、化粧料用組成物の形態がジェルの場合は１〜５０重量％程度、形態が乳液の場合は１〜５０重量％程度、形態がクリームの場合は１〜５０重量％程度である。 The mixing ratio of the eggshell membrane solubilized product to the cosmetic composition is arbitrary, but the cosmetic composition is appropriately selected within a range in which the effects derived from the eggshell membrane solubilized product such as a moisturizing action are expressed. . The ratio of the solubilized eggshell membrane to the cosmetic composition depends on the ratio of the solvent in the solubilized eggshell membrane, the type of the solvent, etc., for example, 1 to 3 when the cosmetic composition is gel. When the form is an emulsion, it is about 1 to 50% by weight. When the form is a cream, it is about 1 to 50% by weight.
本発明の化粧料用組成物は、慣用の化粧料基材を適宜配合し、所望の剤型とすることができる。その形態は特に制限はないが、ジェル、乳液、クリーム等の形態が挙げられる。 The cosmetic composition of the present invention can be formulated into a desired dosage form by appropriately blending a conventional cosmetic base material. The form is not particularly limited, and examples thereof include gels, emulsions, and creams.
化粧料基材としては、目的とする剤型に合わせて、水、水溶性有機溶媒、油脂、ロウ、高級脂肪酸、高級アルコール、エステル類等の従来公知の化粧料基材が適宜選択される。 As the cosmetic base material, conventionally known cosmetic base materials such as water, water-soluble organic solvents, fats and oils, waxes, higher fatty acids, higher alcohols, and esters are appropriately selected according to the intended dosage form.
また、本発明の化粧料用組成物には、本発明の効果を損なわない範囲で通常化粧料や皮膚外用医薬で使用される任意の成分を添加することができる。かかる任意成分の具体例としては、増粘・ゲル化剤、酸化防止剤、紫外線吸収剤、色素剤、金属封鎖剤、防腐剤、ｐＨ調整剤、香料、ミツロウ等が挙げられる。これら任意成分の配合割合は、その目的に応じて適宜選択して決定することができる。 Moreover, the arbitrary components normally used by cosmetics and a skin external medicine can be added to the composition for cosmetics of this invention in the range which does not impair the effect of this invention. Specific examples of such optional components include thickening / gelling agents, antioxidants, ultraviolet absorbers, pigments, metal sequestering agents, preservatives, pH adjusters, fragrances, beeswax and the like. The blending ratio of these optional components can be selected and determined as appropriate according to the purpose.
本発明の化粧料用組成物は、当業者が通常用いる方法によって製造することができ、卵殻膜可溶化物の配合量、配合方法、配合時期は適宜選択することができる。 The cosmetic composition of the present invention can be produced by a method commonly used by those skilled in the art, and the blending amount, blending method, and blending time of the solubilized eggshell membrane can be appropriately selected.
本発明の化粧料用組成物は、スキンケア化粧料用、浴用化粧料用、毛髪化粧料用又はメイクアップ化粧料用等に用いることができるが、好適な用途としては、まゆげ用化粧品、まつげ用化粧品、ネイル用品、毛髪化粧料用組成物（ヘアケア製品）等が挙げられる。 The cosmetic composition of the present invention can be used for skin care cosmetics, bath cosmetics, hair cosmetics, makeup cosmetics, and the like. Suitable applications include eyelash cosmetics and eyelashes. Cosmetics, nail products, compositions for hair cosmetics (hair care products) and the like can be mentioned.
まゆげ用化粧品としては、例えば、アイブロウペンシル、アイブロウマスカラ、アイブロウパウダー等が挙げられる。 Examples of eyebrow cosmetics include eyebrow pencil, eyebrow mascara, eyebrow powder and the like.
まつげ用化粧品としては、例えば、まつげエクステ用グルー、まつげエクステ用リムーバー、まつげエクステ用前処理剤、まつげ美容液、まつげエクステ用コーティング剤、マスカラ、マスカラ下地、吸着剤、クレンジング、アイライナー等が挙げられる。 Examples of cosmetics for eyelashes include eyelash extension glue, eyelash extension remover, eyelash extension pre-treatment agent, eyelash cosmetic, eyelash extension coating, mascara, mascara base, adsorbent, cleansing, eyeliner, etc. It is done.
ネイル用品としては、例えば、プレプライマー、クリアジェル、カラージェル、クレンザー、リムーバー、ベースジェル、トップジェル、キューティクルオイル、キューティクルリムーバー、ポリッシュ、ネイルトリートメント等が挙げられる。 Examples of the nail products include pre-primer, clear gel, color gel, cleanser, remover, base gel, top gel, cuticle oil, cuticle remover, polish, and nail treatment.
以下、本発明の化粧料用組成物の好適な用途の一つである毛髪化粧料用組成物について説明する。 Hereinafter, the composition for hair cosmetics which is one of the suitable uses of the cosmetic composition of the present invention will be described.
（毛髪化粧料用組成物）
本発明の卵殻膜可溶化物組成物は、毛髪化粧料組成物として好適に使用できる。本発明の卵殻膜可溶化物は、従来の卵殻膜可溶化物と比較して分子量が大きいため、低分子化した卵殻膜可溶化物では十分に補修、保護効果が発揮されないようなダメージの大きい毛髪に対しても、十分な補修、保護効果が発揮されやすい。
(Composition for hair cosmetics)
The eggshell membrane solubilized composition of the present invention can be suitably used as a hair cosmetic composition. The eggshell membrane solubilized product of the present invention has a large molecular weight as compared with the conventional eggshell membrane solubilized product, so that the low molecular weight eggshell membrane solubilized product is sufficiently damaged and has a large damage that does not exert its protective effect. Sufficient repair and protection effects are easily exerted on the hair.
毛髪化粧料用組成物としては、例えば、染毛剤やパーマ剤、シャンプー、ヘアリンス、ヘアコンディショナー、ヘアトリートメント、ヘアクリーム、ヘアジェル、ヘアムース、ヘアオイル、スタイリング剤、カーリング剤、縮毛矯正剤、頭髪用カールジェル、育毛剤等が挙げられる。なお、本発明において、毛髪化粧料用組成物には、化粧品のみならず医薬部外品も含む。
すなわち、本発明の毛髪化粧料用組成物は、染色やパーマ、トリートメント、毛髪ケア等の毛髪処理に用いることができる。
Examples of hair cosmetic compositions include hair dyes, permanents, shampoos, hair rinses, hair conditioners, hair treatments, hair creams, hair gels, hair mousses, hair oils, styling agents, curling agents, hair straighteners, and head hair. Examples include curl gel and hair restorer. In the present invention, the composition for hair cosmetics includes not only cosmetics but also quasi drugs.
That is, the composition for hair cosmetics of the present invention can be used for hair treatment such as dyeing, perm, treatment, and hair care.
毛髪化粧料用組成物への卵殻膜可溶化物の配合割合は任意であるが、毛髪化粧料用組成物が、髪のダメージの保護、補修効果等の卵殻膜可溶化物に由来する作用を発現する範囲で適宜選択される。卵殻膜可溶化物の毛髪化粧料用組成物に対する割合は、卵殻膜可溶化物における溶媒の割合や溶媒の種類等に応じて適宜決定されるが、卵殻膜可溶化物の含有量は、毛髪化粧料用組成物の全体に対して１〜５０重量％程度である。 The composition ratio of the solubilized eggshell membrane to the composition for hair cosmetics is arbitrary, but the composition for hair cosmetics has the effects derived from the solubilized eggshell membrane such as protection of hair damage and repair effect. It is appropriately selected within the range of expression. The ratio of the solubilized eggshell membrane to the composition for hair cosmetics is appropriately determined according to the ratio of the solvent in the solubilized eggshell membrane, the type of the solvent, etc. The content of the solubilized eggshell membrane is the hair It is about 1-50 weight% with respect to the whole composition for cosmetics.
本発明の毛髪化粧料用組成物は、慣用の化粧料基材を適宜配合し、所望の剤型とすることができる。その形態は特に制限はないが、例えば、液状、乳液状、クリーム状、ジェル状、ペースト状、ムース状、エアゾール等の形態が挙げられる。 The composition for hair cosmetics of the present invention can be formulated into a desired dosage form by appropriately blending conventional cosmetic base materials. The form is not particularly limited, and examples thereof include forms such as liquid, emulsion, cream, gel, paste, mousse, and aerosol.
化粧料基材としては、目的とする剤型に合わせて、水、水溶性有機溶媒、油脂、ロウ、高級脂肪酸、高級アルコール、エステル類等の従来公知の化粧料基材が適宜選択される。 As the cosmetic base material, conventionally known cosmetic base materials such as water, water-soluble organic solvents, fats and oils, waxes, higher fatty acids, higher alcohols, and esters are appropriately selected according to the intended dosage form.
なお、毛髪化粧料用組成物は、本発明の卵殻膜タンパク質可溶化物以外の卵殻膜タンパク質可溶化物を含有してもよいし、卵殻膜タンパク質を含有してもよい。
また、本発明の化粧料用組成物には、本発明の効果を損なわない範囲で通常使用される任意の成分を添加することができる。かかる任意成分の具体例としては、還元剤、界面活性剤、増粘・ゲル化剤、アルカリ剤、酸化剤、乳化安定剤、酸化防止剤、紫外線吸収剤、色素剤、金属封鎖剤、防腐剤、ｐＨ調整剤、香料、保湿剤、油剤、ミツロウ、シリコーン、キレート剤、着色料等が挙げられる。これら任意成分の配合割合は、その目的に応じて適宜選択して決定することができる。
In addition, the composition for hair cosmetics may contain eggshell membrane protein solubilized products other than the eggshell membrane protein solubilized product of this invention, and may contain eggshell membrane protein.
Moreover, the arbitrary components normally used can be added to the cosmetic composition of this invention in the range which does not impair the effect of this invention. Specific examples of such optional components include reducing agents, surfactants, thickening / gelling agents, alkali agents, oxidizing agents, emulsion stabilizers, antioxidants, ultraviolet absorbers, coloring agents, sequestering agents, preservatives. , PH adjusters, fragrances, moisturizers, oils, beeswax, silicone, chelating agents, coloring agents and the like. The blending ratio of these optional components can be selected and determined as appropriate according to the purpose.
本発明の毛髪化粧料用組成物は、当業者が通常用いる方法によって製造することができ、本発明の卵殻膜タンパク質可溶化物の配合量、配合方法、配合時期は適宜選択することができる。 The composition for hair cosmetics of the present invention can be produced by a method commonly used by those skilled in the art, and the blending amount, blending method, and blending time of the eggshell membrane protein solubilized product of the present invention can be appropriately selected.
（医薬品用組成物）
本発明の組成物は、医薬品用組成物の用途に使用してもよい。なお、本発明の医薬品用組成物は、医薬品のみならず、医薬部外品に配合してもよい。
本発明の医薬品用組成物の形態としては特に限定はないが、一般に皮膚に塗布する形の皮膚外用剤として用いられる場合には、液状やクリーム状である。この場合、医薬品用組成物は、必要に応じて、通常医薬品、医薬部外品に配合される、油性成分、可溶化剤、保湿剤、色素、乳化剤、増粘剤、香料等の任意の成分を含有することができる。
(Pharmaceutical composition)
You may use the composition of this invention for the use of the composition for pharmaceuticals. In addition, you may mix | blend the pharmaceutical composition of this invention not only with a pharmaceutical but a quasi-drug.
The form of the pharmaceutical composition of the present invention is not particularly limited, but when used as a skin external preparation generally applied to the skin, it is liquid or creamy. In this case, the pharmaceutical composition is an optional component such as an oily component, a solubilizer, a moisturizer, a dye, an emulsifier, a thickener, a fragrance, etc. Can be contained.
また、本発明の医薬品用組成物は、入浴剤、ボディーソープ、シャンプー等の入浴用組成物に配合して用いてもよい。剤型としては、一般に用いられる、水溶液、Ｗ／Ｏ型又はＯ／Ｗ型エマルション、適当な賦形剤等を用いて顆粒剤その他の粉末、錠剤等としてもよい。 Further, the pharmaceutical composition of the present invention may be used by blending with a bath composition such as a bath agent, body soap, shampoo and the like. The dosage form may be a granule or other powder, tablet or the like using a commonly used aqueous solution, W / O type or O / W type emulsion, an appropriate excipient and the like.
また、本発明の医薬品用組成物は、人体に対する毒性や刺激性が少なく、安全であるため、外用剤としてのみならず、服用剤として経口投与とすることもできる。
経口投与に利用される剤形としては、具体的には、固形製剤として、粉末剤、顆粒剤、錠剤、カプセル剤、トローチ等が挙げられる。また、液状製剤として内用液剤、外用液剤、懸濁剤、乳剤、シロップ剤等が例示され、これら剤形やその他の剤形が目的に応じて適宜選択される。また、経口投与用の医薬品組成物として、薬学的に許容される通常の担体、結合剤、安定化剤、賦形剤、希釈剤、ｐＨ緩衝剤、崩壊剤、可溶化剤、溶解補助剤、等張剤等の各種調剤用配合成分をさらに適宜含有していてもよい。
Moreover, since the pharmaceutical composition of the present invention is safe and less toxic and irritating to the human body, it can be administered not only as an external preparation but also as an oral administration.
Specific examples of dosage forms utilized for oral administration include powders, granules, tablets, capsules, troches and the like as solid preparations. Examples of liquid preparations include internal liquids, external liquids, suspensions, emulsions, syrups and the like, and these and other dosage forms are appropriately selected according to the purpose. In addition, as pharmaceutical compositions for oral administration, pharmaceutically acceptable ordinary carriers, binders, stabilizers, excipients, diluents, pH buffers, disintegrants, solubilizers, solubilizers, Various preparation blending components such as isotonic agents may be further contained as appropriate.
本発明の医薬品組成物は、当業者が通常用いる方法によって製造することができ、卵殻膜タンパク質可溶化物の配合量、配合方法、配合時期は適宜選択することができる。医薬品組成物の投与量は、対象者の性別や年齢、体重、投与形態等に応じて適宜選定することができる。 The pharmaceutical composition of the present invention can be produced by a method commonly used by those skilled in the art, and the blending amount, blending method, and blending time of the eggshell membrane protein solubilized product can be appropriately selected. The dosage of the pharmaceutical composition can be appropriately selected according to the sex, age, weight, dosage form, etc. of the subject.
（機能性食品）
一方、日常的に飲食することで、本発明の卵殻膜タンパク質可溶化物を摂取したい場合には、該食品、飲料に含有させて機能性食品としてもよい。
ここでいう「機能性食品」とは、一般食品に加えて、健康食品、栄養補助食品、栄養機能食品、栄養保険食品等、健康の維持の目的で摂取する食品及び／又は飲料を意味している。なお、機能性食品として製品化する場合には、食品に用いられる様々な添加剤、具体的には、着色料、保存料、増粘安定剤、酸化防止剤漂白剤、防菌防黴剤、酸味料、調味料、乳化剤、強化剤、製造用剤、香料等を添加していてもよい。
(Functional food)
On the other hand, when it is desired to ingest the eggshell membrane protein solubilized product of the present invention by eating and drinking on a daily basis, it may be contained in the food or beverage to obtain a functional food.
The term “functional food” as used herein means foods and / or beverages ingested for the purpose of maintaining health, such as health foods, dietary supplements, nutritional functional foods, nutrition insurance foods, in addition to general foods. Yes. In addition, when commercializing as a functional food, various additives used in food, specifically, coloring agents, preservatives, thickening stabilizers, antioxidant bleaching agents, antibacterial and antifungal agents, A sour agent, a seasoning, an emulsifier, a reinforcing agent, a manufacturing agent, a fragrance and the like may be added.
本発明の機能性食品の対象となる、食品、飲料は特に限定されるものではない。例えば、食品として、ソーセージ、ハム、魚介加工品、ゼリー、キャンディー、チューインガム等の食品類が挙げられる。また、飲料としては、各種の茶類、清涼飲料水、酒類、栄養ドリンク等が挙げられる。本発明の卵殻膜可溶化物は、このような食品、飲料に添加することにより、簡易に経口摂取することができる。また、卵殻膜可溶化物の配合量は、食品、飲料の種類、目的とする作用に応じて適宜決定すればよい。 The foods and beverages that are the targets of the functional food of the present invention are not particularly limited. Examples of food include foods such as sausage, ham, processed seafood, jelly, candy, and chewing gum. Examples of the beverage include various teas, soft drinks, alcoholic beverages, and energy drinks. The eggshell membrane solubilized product of the present invention can be easily taken orally by adding to such foods and beverages. Moreover, what is necessary is just to determine the compounding quantity of eggshell membrane solubilized material suitably according to the kind of food and drink, and the objective effect | action.
以下に実施例を挙げて本発明をより具体的に説明するが、本発明はこれらに限定されるものではない。 Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples.
「実施例１」
（１．卵殻膜の乳酸による前処理）
水９４．４重量部及びＬ−乳酸（和光純薬工業（株）、濃度８５〜９２％）５．６重量部の乳酸溶液に、粉末状の卵殻膜１０重量部を添加し、常温（約２５℃）下、１２時間撹拌し、卵殻膜に含まれるカルシウム成分を溶解させた。なお、撹拌後の混合液のｐＨをｐＨメーター（東亜ディーケーケー製、ＨＭ−３０Ｒ）で測定したところ、ｐＨ３．１９であった。次いで、撹拌後の混合液を、ろ過器を用い、固形物と液体に分離した。得られた固形物を４０℃で減圧乾燥し、乾燥物を得た。最終的に得られた固形物（卵殻膜タンパク質）は、３．１重量部であった。この固形物のタンパク質量をケルダール法により評価したところ、タンパク質の割合が９４．２重量％であった。得られた卵殻膜は、カルシウム顔料にして０．５％以下の高純度であった。
"Example 1"
(1. Pretreatment of eggshell membranes with lactic acid)
To a lactic acid solution of 94.4 parts by weight of water and 5.6 parts by weight of L-lactic acid (Wako Pure Chemical Industries, Ltd., concentration 85-92%), 10 parts by weight of a powdered eggshell membrane is added at room temperature (about The mixture was stirred for 12 hours at 25 ° C. to dissolve the calcium component contained in the eggshell membrane. In addition, it was pH 3.19 when pH of the liquid mixture after stirring was measured with the pH meter (the product made by Toa DK, HM-30R). Next, the mixed liquid after stirring was separated into a solid and a liquid using a filter. The obtained solid was dried under reduced pressure at 40 ° C. to obtain a dried product. The finally obtained solid (eggshell membrane protein) was 3.1 parts by weight. When the protein amount of this solid was evaluated by the Kjeldahl method, the protein ratio was 94.2% by weight. The obtained eggshell membrane was a calcium pigment having a high purity of 0.5% or less.
（２．卵殻膜タンパク質の可溶化処理）
前処理後の卵殻膜タンパク質の可溶化処理は、以下の手順で行った。
まず、純水に水酸化ナトリウムを添加して混合し、水酸化ナトリウム濃度１重量％の処理溶液を調整した。次いで、水酸化ナトリウム濃度１重量％の処理溶液９５重量部に対し、卵殻膜タンパク質５重量部を添加し、均一になるように撹拌し、懸濁液を得た。この懸濁液のｐＨをｐＨメータ（東亜ディーケーケー製、ＨＭ−３０Ｒ）で測定したところ、ｐＨ１２以上であった。
さらに、１４０℃まで１分以内に到達するように直接蒸気吹き込み式連続殺菌装置（イズミフードマシナリー製スチームインジェクションＳＤＩ―１５０殺菌テスト機）を用いて、１４０℃まで加熱を行い、同温度で２０秒間保持し、卵殻膜タンパク質を可溶化し、卵殻膜タンパク質可溶化物を得た。可溶化処理後、室温まで冷却し、実施例１の卵殻膜タンパク質の可溶化物を得た。
(2. Solubilization of eggshell membrane protein)
The solubilization treatment of the eggshell membrane protein after the pretreatment was performed according to the following procedure.
First, sodium hydroxide was added to pure water and mixed to prepare a treatment solution having a sodium hydroxide concentration of 1% by weight. Next, 5 parts by weight of eggshell membrane protein was added to 95 parts by weight of the treatment solution having a sodium hydroxide concentration of 1% by weight, and the mixture was stirred uniformly to obtain a suspension. The pH of this suspension was measured with a pH meter (manufactured by Toa DKK Co., HM-30R) and found to be pH 12 or higher.
Furthermore, using a direct steam blow type continuous sterilization device (Izumi Food Machinery's steam injection SDI-150 sterilization test machine) to reach 140 ° C. within 1 minute, it is heated to 140 ° C. and kept at the same temperature for 20 seconds. The eggshell membrane protein was solubilized to obtain a solubilized eggshell membrane protein. After the solubilization treatment, the mixture was cooled to room temperature to obtain a solubilized product of eggshell membrane protein of Example 1.
「実施例２」
水酸化ナトリウム濃度１重量％の処理溶液に代えて、水酸化ナトリウム濃度２重量％の処理溶液を用いた以外は、実施例１と同様にして卵殻膜タンパク質の可溶化物を得た。
"Example 2"
A solubilized egg shell membrane protein was obtained in the same manner as in Example 1 except that a treatment solution having a sodium hydroxide concentration of 2% by weight was used instead of the treatment solution having a sodium hydroxide concentration of 1% by weight.
「実施例３」
水酸化ナトリウム濃度１重量％の処理溶液に代えて、水酸化ナトリウム濃度３重量％の処理溶液を用いた以外は、実施例１と同様にして卵殻膜タンパク質の可溶化物を得た。
"Example 3"
A solubilized egg shell membrane protein was obtained in the same manner as in Example 1 except that a treatment solution having a sodium hydroxide concentration of 3% by weight was used instead of the treatment solution having a sodium hydroxide concentration of 1% by weight.
「比較例１」
水酸化ナトリウム濃度３重量％の処理を用いて、卵殻膜タンパク質の懸濁液を調整した。この懸濁液のｐＨをｐＨメータ（東亜ディーケーケー製、ＨＭ−３０Ｒ）で測定したところ、ｐＨ１２以上であった。
さらに、１００℃まで２０分かけて昇温し、同温度で３時間保持し、卵殻膜タンパク質を可溶化し、卵殻膜タンパク質可溶化物を得た。可溶化処理後、室温まで冷却し、比較例１の卵殻膜タンパク質の可溶化物を得た。
"Comparative Example 1"
A suspension of eggshell membrane protein was prepared using a treatment with a sodium hydroxide concentration of 3% by weight. The pH of this suspension was measured with a pH meter (manufactured by Toa DKK Co., HM-30R) and found to be pH 12 or higher.
Further, the temperature was raised to 100 ° C. over 20 minutes and held at the same temperature for 3 hours to solubilize the eggshell membrane protein and obtain a solubilized eggshell membrane protein. After the solubilization treatment, the mixture was cooled to room temperature to obtain a solubilized egg shell membrane protein of Comparative Example 1.
「評価」
（１）卵殻膜タンパク質可溶化物の重量平均分子量
実施例１〜３の卵殻膜タンパク質可溶化物及び比較例１の卵殻膜タンパク質可溶化物について、サイズ排除クロマトグラフィーを測定した。なお、卵殻膜タンパク質可溶化物は、下記試料濃度に調整後、精密ろ過フィルター（ザルトリウス製ミニザルトＲＣ１５、孔径０．２μｍ）を通して、装置に注入した。得られたサイズ排除クロマトグラムを図１〜４に示す。
また、サイズ排除クロマトグラフィーで得られた分子量分布から重量平均分子量を算出した。結果を表１に示す。
"Evaluation"
(1) Weight average molecular weight of solubilized eggshell membrane protein Size exclusion chromatography was measured for the solubilized eggshell membrane protein of Examples 1 to 3 and the solubilized eggshell membrane protein of Comparative Example 1. In addition, the eggshell membrane protein lysate was adjusted to the following sample concentration, and then injected into the apparatus through a microfiltration filter (Sartorius Mini-Salto RC15, pore size 0.2 μm). The obtained size exclusion chromatograms are shown in FIGS.
Further, the weight average molecular weight was calculated from the molecular weight distribution obtained by size exclusion chromatography. The results are shown in Table 1.
なお、サイズ排除クロマトグラフィーの条件は以下のとおりである；
株式会社島津製作所社製高速液体クロマトグラフ、装置名「プロミネンス」
カラム：ＧＥ製、商品名「Ｓｕｐｅｒｄｅｘ ｐｅｐｔｉｄｅ ＨＲ １０／３００」１本
溶離液：３０％アセトニトリル―７０％水−０．１％トリフルオロ酢酸
流量：０．５ｍｌ／ｍｉｎ
カラム温度：４０℃
検出器：紫外吸光光度計（測定波長２１４ｎｍ）、
試料濃度：５〜１０ｍｇ／ｍｌ
試料注入量：２５μＬ
The size exclusion chromatography conditions are as follows:
High-performance liquid chromatograph manufactured by Shimadzu Corporation, apparatus name “Prominence”
Column: GE, trade name “Superdex peptide HR 10/300” 1 eluent: 30% acetonitrile-70% water-0.1% trifluoroacetic acid flow rate: 0.5 ml / min
Column temperature: 40 ° C
Detector: ultraviolet absorptiometer (measurement wavelength 214 nm),
Sample concentration: 5-10 mg / ml
Sample injection volume: 25 μL
（２）システイン及びシスチンの含有量
実施例１〜３の卵殻膜タンパク質可溶化物及び比較例１の卵殻膜タンパク質可溶化物について、アミノ酸組成分析によりシステイン及びシスチンの含有量を算出した。
なお、アミノ酸組成分析は、酸化剤であるアジ化ナトリウム存在下塩酸加水分解してシステイン及びシスチンについてはシステイン酸の形にし、AccQTag誘導体化（日本ウォーターズ株式会社）して超高圧高速液体クロマトグラフィー（日本ウォーターズ株式会社製UPLC H class）で分析した。
また、システイン及びシスチンの含有量は、原料となる可溶化処理前の卵殻膜タンパク質のシステイン及びシスチンの含有量に対する残存率として求めた。すなわち、残存率は以下の式で表される。
残存率（％） ＝ （卵殻膜タンパク質可溶化物のシステイン及びシスチンの含有量／可溶化処理前の卵殻膜タンパク質のシステイン及びシスチンの含有量）×１００
結果を表２に示す。
(2) Content of cysteine and cystine About the eggshell membrane protein solubilized product of Examples 1-3 and the eggshell membrane protein solubilized product of Comparative Example 1, the content of cysteine and cystine was calculated by amino acid composition analysis.
The amino acid composition analysis was performed by hydrolyzing hydrochloric acid in the presence of an oxidizing agent, sodium azide, to form cysteine and cystine in the form of cysteic acid, AccQTag derivatization (Nippon Waters Co., Ltd.), and ultrahigh pressure high performance liquid chromatography ( Analysis was carried out with UPLC H class manufactured by Nippon Waters Co., Ltd.
The cysteine and cystine contents were determined as the residual ratio of the eggshell membrane protein before the solubilization treatment as a raw material to the cysteine and cystine contents. That is, the remaining rate is expressed by the following formula.
Residual rate (%) = (content of cysteine and cystine in solubilized eggshell membrane protein / content of cysteine and cystine in eggshell membrane protein before solubilization) × 100
The results are shown in Table 2.
図１に示すように、実施例１の卵殻膜タンパク質可溶化物は、上記測定条件でサイズ排除クロマトグラフィーを測定し得られたサイズ排除クロマトグラムおいて、保持時間１５分にピークＡを有し、また、総ピーク面積(保持時間０分から、実質的にピーク強度及びピークの傾きが０となる保持時間４５分までの間のピークの面積の和）に対する、前記ピークＡの面積の比が１５．５％である。
また、図２に示すように、実施例２の卵殻膜タンパク質可溶化物は、上記測定条件でのサイズ排除クロマトグラムおいて、保持時間１５分にピークＡを有し、また、総ピーク面積に対する、前記ピークＡの面積の比が１０．８％である。
また、図３に示すように、実施例３の卵殻膜タンパク質可溶化物は、上記測定条件でのサイズ排除クロマトグラムおいて、保持時間１５分にピークＡを有し、また、総ピーク面積に対する、前記ピークＡの面積の比が９．４％である。
As shown in FIG. 1, the solubilized eggshell membrane protein of Example 1 has a peak A at a retention time of 15 minutes in a size exclusion chromatogram obtained by measuring size exclusion chromatography under the above measurement conditions. Further, the ratio of the area of the peak A to the total peak area (the sum of the areas of the peaks from the holding time of 0 minutes to the holding time of 45 minutes where the peak intensity and the peak slope are substantially zero) is 15 .5%.
Moreover, as shown in FIG. 2, the eggshell membrane protein lysate of Example 2 has a peak A at a retention time of 15 minutes in the size exclusion chromatogram under the above-described measurement conditions, and it corresponds to the total peak area. The area ratio of the peak A is 10.8%.
Moreover, as shown in FIG. 3, the eggshell membrane protein lysate of Example 3 has a peak A at a retention time of 15 minutes in the size exclusion chromatogram under the above measurement conditions, The area ratio of the peak A is 9.4%.
一方、図４に示すように、比較例１の卵殻膜可溶化物は、上記測定条件でのサイズ排除クロマトグラムにおいて、保持時間１６分より短い時間にピークは検出されなかった。 On the other hand, as shown in FIG. 4, no peak was detected in the solubilized eggshell membrane of Comparative Example 1 in a time shorter than the retention time of 16 minutes in the size exclusion chromatogram under the above measurement conditions.
また、実施例１〜３の卵殻膜タンパク質可溶化物と比較例１の卵殻膜タンパク質可溶化物の重量平均分子量の比較からも、本発明の卵殻膜タンパク質の可溶化物は高分子量成分を多く含むことがわかる。 Moreover, from the comparison of the weight average molecular weights of the eggshell membrane protein lysate of Examples 1 to 3 and the eggshell membrane protein lysate of Comparative Example 1, the lysate of eggshell membrane protein of the present invention has many high molecular weight components. I understand that it contains.
また、表２に示すように、実施例１〜３の卵殻膜タンパク質可溶化物は、比較例１の卵殻膜タンパク質可溶化物と比較して、原料に対するシステイン及びシスチンの含有量の残存率が多く、本製造方法では、システイン及びシスチンが分解しにくいことが分かる。そのため、得られた卵殻膜タンパク質可溶化物は、まゆげ用化粧品やまつげ用化粧品、ネイル用品、毛髪化粧料用組成物の原料として用いることで、高い保護効果が期待できる。 Moreover, as shown in Table 2, compared with the eggshell membrane protein solubilized product of Comparative Example 1 in the eggshell membrane protein solubilized products of Examples 1 to 3, the residual ratio of the content of cysteine and cystine relative to the raw material In many cases, it can be seen that cysteine and cystine are difficult to decompose in this production method. Therefore, the obtained eggshell membrane protein solubilized product can be expected to have a high protective effect when used as a raw material for eyebrow cosmetics, eyelash cosmetics, nail products, and hair cosmetic compositions.
このように、実施例１〜３の卵殻膜タンパク質の可溶化物は、従来の卵殻膜タンパク質可溶化物と比較して高分子量を多く含むにもかかわらず、水に可溶化することができ、システイン及びシスチンも多く含有しており、従来の卵殻膜タンパク質可溶化物とは異なる性質を有したものである。 Thus, the solubilized eggshell membrane protein of Examples 1 to 3 can be solubilized in water, despite the high molecular weight compared to the conventional eggshell membrane protein solubilized product, It contains a large amount of cysteine and cystine, and has properties different from those of the conventional solubilized eggshell membrane protein.
本発明によれば、化粧料用組成物、医療用組成物、機能性食品の原料として好適に用いられる卵殻膜タンパク質可溶化物及び、該卵殻膜タンパク質を低コストに生産することができる製造方法が提供されるので、工業的に有望である。 According to the present invention, an eggshell membrane protein solubilized product suitably used as a raw material for cosmetic compositions, medical compositions, and functional foods, and a production method capable of producing the eggshell membrane protein at low cost Is promising industrially.
Claims (8)
ピークトップを保持時間１５．０〜１５．２分の間に持つピークＡを有し、総ピーク面積に対する、前記ピークＡの面積の比が９％以上１５．５％以下であり、
かつ、卵殻膜タンパク質可溶化物の重量平均分子量が、９０００以上１４０００以下であり、システイン及びシスチンの含有量が１．５質量％以上３．３質量％以下である、
卵殻膜タンパク質可溶化物。
（測定条件）
測定方法：サイズ排除クロマトグラフィー
カラム：ＧＥ製、商品名「Ｓｕｐｅｒｄｅｘ ｐｅｐｔｉｄｅ ＨＲ １０／３００」１本
溶離液：３０％アセトニトリル―７０％水−０．１％トリフルオロ酢酸
流量：０．５ｍｌ／ｍｉｎ
カラム温度：４０℃
検出器：紫外吸光光度計（測定波長２１４ｎｍ）
試料濃度：５〜１０ｍｇ／ｍｌ
試料注入量：２５μＬ In the size exclusion chromatogram measured under the following conditions:
It has a peak A having a peak top with a retention time of 15.0 to 15.2 minutes, and the ratio of the area of the peak A to the total peak area is 9% to 15.5%,
And, the weight average molecular weight of the solubilized eggshell membrane protein is 9000 or more and 14000 or less, and the content of cysteine and cystine is 1.5 mass% or more and 3.3 mass% or less.
Egg shell membrane protein lysate.
(Measurement condition)
Measurement method: Size exclusion chromatography column: GE, trade name “Superdex peptide HR 10/300” 1 eluent: 30% acetonitrile-70% water-0.1% trifluoroacetic acid flow rate: 0.5 ml / min
Column temperature: 40 ° C
Detector: UV absorptiometer (measurement wavelength 214 nm)
Sample concentration: 5-10 mg / ml
Sample injection volume: 25 μL
アルカリ条件に調整した卵殻膜タンパク質の懸濁液を処理温度１３０℃以上１６０℃以下で、かつ、当該処理温度に５分以内に到達するよう加熱する工程を有する卵殻膜タンパク質可溶化物の製造方法。 A method for producing a solubilized eggshell membrane protein according to claim 1,
A method for producing a solubilized eggshell membrane protein comprising a step of heating a suspension of eggshell membrane protein adjusted to an alkaline condition at a treatment temperature of 130 ° C to 160 ° C so as to reach the treatment temperature within 5 minutes. .
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