$\gamma$$\gamma$gamma\gamma-Poly Glutamic Acid: A Novel Peptide for Skin Care
$\gamma$$\gamma$gamma\gamma -聚谷氨酸：一種用於皮膚護理的新型肽

$\gamma$$\gamma$gamma\gamma-Poly glutamic acid ( $\gamma$$\gamma$gamma\gamma-PGA) is a novel molecule that is a component of the mucilage of the fermented soybean food product commonly found in Japan called natto. The International Nomenclature Cosmetic Ingredient (INCI) name of $\gamma$$\gamma$gamma\gamma-PGA is natto gum and it is classified as a film-forming agent. The sodium salt of $\gamma$$\gamma$gamma\gamma-PGA, which is more commonly used and thus served as the test sample in this paper, displays the visual appearance of a sticky paste as illustrated in Figure 1.
$\gamma$$\gamma$gamma\gamma -聚谷氨酸 （ $\gamma$$\gamma$gamma\gamma -PGA） 是一種新型分子，是日本常見的發酵大豆食品納豆的粘液成分。-PGA 的國際 $\gamma$$\gamma$gamma\gamma 命名化妝品成分 （INCI） 名稱是納豆膠，它被歸類為成膜劑。-PGA 鈉 $\gamma$$\gamma$gamma\gamma 鹽是更常用的，因此在本文中用作測試樣品，如圖 1 所示，它顯示出粘稠糊狀物的視覺外觀。

The fermented soybean mucilage consists of a mixture of $\gamma$$\gamma$gamma\gamma-PGA and fructan produced by Bacillus natto. ${}^{\text{t.}2}$${}^{\text{t. }2}$^("t. "2){ }^{\text {t. } 2} Further research by Bovarnick revealed $\gamma$$\gamma$gamma\gamma-PGA to be a fermentation by-product freely secreted into growth medium outside of the cell walls of Bacillus subtilis. ${}^3$${}^3$^(3){ }^{3} This discovery advanced collateral investigations to determine whether this bacteriological process could be reproduced in other species of Bacillus. ${}^{411}$${}^{411}$^(411){ }^{411}
發酵大豆粘液由 $\gamma$$\gamma$gamma\gamma -PGA 和納豆芽孢桿菌產生的果聚糖混合物組成。 ${}^{\text{t.}2}$${}^{\text{t. }2}$^("t. "2){ }^{\text {t. } 2} Bovarnick 的進一步研究表明 $\gamma$$\gamma$gamma\gamma ，-PGA 是一種發酵副產物，可自由分泌到枯草芽孢桿菌細胞壁外的生長培養基中。 ${}^3$${}^3$^(3){ }^{3} 這一發現推進了附帶調查，以確定這種細菌學過程是否可以在其他種類的芽孢桿菌中複製。 ${}^{411}$${}^{411}$^(411){ }^{411}

Recent work by Lung and Da-Yeh further illustrates the applications and production of $\gamma$$\gamma$gamma\gamma-PGA from microorganisms by gamma irradiating to promote chemical cross-linking between monomeric units to form a $\gamma$$\gamma$gamma\gamma-PGA hydrogel. ${}^{12}$${}^{12}$^(12){ }^{12}
Lung 和 Da-Yeh 最近的工作進一步說明了通過伽馬輻照從微生物中生產 $\gamma$$\gamma$gamma\gamma -PGA 的應用和生產，以促進單體單元之間的化學交聯形成 $\gamma$$\gamma$gamma\gamma -PGA 水凝膠。 ${}^{12}$${}^{12}$^(12){ }^{12}

The finished gel was lyophilized to assure its physical chemical stability.
將成品凝膠凍乾以確保其物理化學穩定性。

The lyophilized product was characterized by Lung and Da-yeh by nuclear
凍幹產物以 Lung 和 Da-yeh 為特徵
magnetic resonance ( ${\mathrm{C}}^{13}$${\mathrm{C}}^{13}$C^(13)\mathrm{C}^{13} and ${\mathrm{H}}^1$${\mathrm{H}}^1$H^(1)\mathrm{H}^{1} ), Fourier Transform Infrared Spectroscopy and thermal gravimetric analysis in its protonated form, as well as the salts of ${\mathrm{Na}}^{+},{\mathrm{K}}^{+},{\mathrm{NH}}_{+}^{-},{\mathrm{Ca}}^{2+}$${\mathrm{Na}}^{+},{\mathrm{K}}^{+},{\mathrm{NH}}_{+}^{-},{\mathrm{Ca}}^{2+}$Na^(+),K^(+),NH_(+)^(-),Ca^(2+)\mathrm{Na}^{+}, \mathrm{K}^{+}, \mathrm{NH}_{+}^{-}, \mathrm{Ca}^{2+} and ${\mathrm{Mg}}^{2+}$${\mathrm{Mg}}^{2+}$Mg^(2+)\mathrm{Mg}^{2+}.
質子化形式的磁共振 （ ${\mathrm{C}}^{13}$${\mathrm{C}}^{13}$C^(13)\mathrm{C}^{13} 和 ${\mathrm{H}}^1$${\mathrm{H}}^1$H^(1)\mathrm{H}^{1} ）、傅里葉變換紅外光譜和熱重分析，以及和 ${\mathrm{Mg}}^{2+}$${\mathrm{Mg}}^{2+}$Mg^(2+)\mathrm{Mg}^{2+} 的 ${\mathrm{Na}}^{+},{\mathrm{K}}^{+},{\mathrm{NH}}_{+}^{-},{\mathrm{Ca}}^{2+}$${\mathrm{Na}}^{+},{\mathrm{K}}^{+},{\mathrm{NH}}_{+}^{-},{\mathrm{Ca}}^{2+}$Na^(+),K^(+),NH_(+)^(-),Ca^(2+)\mathrm{Na}^{+}, \mathrm{K}^{+}, \mathrm{NH}_{+}^{-}, \mathrm{Ca}^{2+} 鹽。

The lyophilized product is a linear homopolypeptide consisting of only glutamic acid residuals composed of both dextro and levo optical isomers. The $\gamma$$\gamma$gamma\gamma-peptide bond linkages exhibit a range of polymerization from 1000-12000 units. Figure 2 depicts the repeating units of $\gamma$$\gamma$gamma\gamma-PGA.
凍幹產物是一種線性同多肽，僅由右旋和左旋光學異構體組成的谷氨酸殘餘物組成。 $\gamma$$\gamma$gamma\gamma -肽鍵的聚合範圍為 1000-12000 個單位。圖 2 描述了 -PGA 的 $\gamma$$\gamma$gamma\gamma 重複單元。

The molecular weight ranged from 100,000 to $4,000,000$$4,000,000$4,000,0004,000,000 Daltons with an assay value above $95.0\mathrm{\%}$$95.0\mathrm{\%}$95.0%95.0 \%. Physicalchemical properties such as the pH
分子量範圍為 100,000 至 $4,000,000$$4,000,000$4,000,0004,000,000 Daltons，測定值高於 $95.0\mathrm{\%}$$95.0\mathrm{\%}$95.0%95.0 \% 。物理化學性質，如 pH 值

Figure 1. The sodium salt of $\gamma -PGA$$\gamma -PGA$gamma-PGA\gamma-P G A displays the visual appearance of a sticky paste.
圖 1.的鈉鹽 $\gamma -PGA$$\gamma -PGA$gamma-PGA\gamma-P G A 顯示粘稠糊狀物的視覺外觀。

Figure 2. Repeating unit of $\gamma$$\gamma$gamma\gamma-PGA structure
圖 2.-PGA 結構的 $\gamma$$\gamma$gamma\gamma 重複單元
buffering capacity are exhibited in a range of 4.0 to 5.0 .
緩衝能力在 4.0 到 5.0 的範圍內顯示。

Viscosity can vary as a function of temperature, pH and concentration to influence performance characteristics. The protonated form of $\gamma$$\gamma$gamma\gamma-PGA is completely soluble in trifluorozincate, dimethyl sulfoxide, hydrazidine and a high concentration of sulfuric acid. The sodium form, which is soluble in water, can assume different molecular conformations depending on pH conditions and binding to other counter ions. Figure 3 illustrates the elucidated structure of the protonated form of $\gamma$$\gamma$gamma\gamma-PGA and Figure 4 depicts the elucidated structure of the sodium salt of $\gamma$$\gamma$gamma\gamma-PGA.
粘度會隨溫度、pH 值和濃度而變化，從而影響性能特徵。-PGA 的 $\gamma$$\gamma$gamma\gamma 質子化形式完全溶於三氟鋅酸鹽、二甲基亞硯、肼和高濃度硫酸。溶於水的鈉形式可以根據 pH 條件呈現不同的分子構象，並與其他反離子結合。圖 3 說明瞭 $\gamma$$\gamma$gamma\gamma -PGA 質子化形式的闡明結構，圖 4 描述了 -PGA 鈉 $\gamma$$\gamma$gamma\gamma 鹽的闡明結構。

The chemistry of $\gamma -\mathrm{PGA}$$\gamma -\mathrm{PGA}$gamma-PGA\gamma-\mathrm{PGA} allows it to be homogenously miscible as well as chemically stable in a matrix of ingredients typically used in facial creams, such as: glycerin, butylene glycol, alcohol, beta-glucan, hydrogenated lecithin, PEG-50, triethanolamine, disodium EDTA, methylparaben, ethylparaben, propylparaben, fragrance (parfum), vitamin E, carbomer, beta carotene, cetyl alcohol and disodium EDTA.
其 $\gamma -\mathrm{PGA}$$\gamma -\mathrm{PGA}$gamma-PGA\gamma-\mathrm{PGA} 化學性質使其在面霜中常用的成分基質中具有均質混溶性和化學穩定性，例如：甘油、丁二醇、酒精、β-葡聚糖、氫化卵磷脂、PEG-50、三乙醇胺、EDTA 二鈉、對羥基苯甲酸甲酯、對羥基苯甲酸乙酯、對羥基苯甲酸丙酯、香料（香水）、維生素 E、卡波姆、β-胡蘿蔔素、鯨蠟醇和 EDTA 二鈉。

Additional qualities of $\gamma -\mathrm{PGA}$$\gamma -\mathrm{PGA}$gamma-PGA\gamma-\mathrm{PGA} include the ability to form a smooth, elastic, self-moisturizing and soft film on the skin, resulting in improved sensory perception and protection of the outer layer of skin. ${}^{13}$${}^{13}$^(13){ }^{13} It is an excellent hydrophilic humectant and is capable of increasing the production of such natural moisturizing factors as pyr-
其他品質 $\gamma -\mathrm{PGA}$$\gamma -\mathrm{PGA}$gamma-PGA\gamma-\mathrm{PGA} 包括能夠在皮膚上形成光滑、有彈性、自保濕和柔軟的薄膜，從而改善感官感知和保護皮膚外層。 ${}^{13}$${}^{13}$^(13){ }^{13} 它是一種極好的親水性保濕劑，能夠增加 pyr-
rolidone carboxylic acid (PCA), lactic acid and urocanic acid, as compared to hyaluronic acid and soluble collagen. ${}^{13}$${}^{13}$^(13){ }^{13}
羅利酮羧酸 （PCA）、乳酸和尿酸，與透明質酸和可溶性膠原蛋白相比。 ${}^{13}$${}^{13}$^(13){ }^{13}

Additional work by Ichimara further supports these findings as illustrated in Figure 5. ${}^{14}$${}^{14}$^(14){ }^{14}
Ichimara 的其他工作進一步支持了這些發現，如圖 5 所示。 ${}^{14}$${}^{14}$^(14){ }^{14}

Figure 3. The elucidated structure of the protonated $\gamma -PGA$$\gamma -PGA$gamma-PGA\gamma-P G A, where $n=H,Na+,Ca++$$n=H,Na+,Ca++$n=H,Na+,Ca++n=H, N a+, C a++, $\mathrm{Mg}++,\mathrm{K}+$$\mathrm{Mg}++,\mathrm{K}+$Mg++,K+\mathrm{Mg}++, \mathrm{K}+.
圖 3.質子化 $\gamma -PGA$$\gamma -PGA$gamma-PGA\gamma-P G A 的闡明結構 ，其中 $n=H,Na+,Ca++$$n=H,Na+,Ca++$n=H,Na+,Ca++n=H, N a+, C a++ ， $\mathrm{Mg}++,\mathrm{K}+$$\mathrm{Mg}++,\mathrm{K}+$Mg++,K+\mathrm{Mg}++, \mathrm{K}+ 。

Figure 4. The elucidated structure of the sodium salt of $\gamma -PGA$$\gamma -PGA$gamma-PGA\gamma-P G A, where $n=H,Na+Ca++$$n=H,Na+Ca++$n=H,Na+Ca++n=H, N a+C a++, Mg++, K+.
圖 4.鈉鹽 $\gamma -PGA$$\gamma -PGA$gamma-PGA\gamma-P G A 的闡明結構，其中 $n=H,Na+Ca++$$n=H,Na+Ca++$n=H,Na+Ca++n=H, N a+C a++ ， Mg++， K+.

Figure 5. Comparison of $\gamma -$$\gamma -$gamma-\gamma--PGA, hyaluronic acid (HA) and soluble collagen in the increase of NMF in the stratum corneum
圖 5. $\gamma -$$\gamma -$gamma-\gamma- -PGA、透明質酸 （HA） 和可溶性膠原蛋白在角質層 NMF 增加中的比較
$\gamma$$\gamma$gamma\gamma-PGA also offers antiaging benefits. A study conducted by Meiji Seika examined the ingredient’s potential as a natural replacement for collagen or hyaluronic acid. ${}^{15}$${}^{15}$^(15){ }^{15} The origin of $\gamma$$\gamma$gamma\gamma-PGA from fermented soybeans and its composition of enzymes, oligosaccharides and vitamins was shown in cosmetic formulations to favorably augment the natural moisturizing factor and elasticity of the skin more than collagen and hyaluronic acid during a 28 -day period, as detailed in Figure 6. The study suggests that $\gamma$$\gamma$gamma\gamma-PGA could successfully replace collagen and hyaluronic acid in skin care products.
$\gamma$$\gamma$gamma\gamma - PGA 還提供抗衰老益處。明治精華進行的一項研究考察了該成分作為膠原蛋白或透明質酸天然替代品的潛力。 ${}^{15}$${}^{15}$^(15){ }^{15} -PGA 來自發酵大豆的 $\gamma$$\gamma$gamma\gamma 來源及其酶、低聚糖和維生素的組成在化妝品配方中顯示，在 28 天內，比膠原蛋白和透明質酸更能增強皮膚的天然保濕因數和彈性，如圖 6 所示。研究表明， $\gamma$$\gamma$gamma\gamma -PGA 可以成功替代護膚品中的膠原蛋白和透明質酸。

As Figure 6 demonstrates, products containing $\gamma$$\gamma$gamma\gamma-PGA reinforce the skin’s support structure to energize cells, stimulating the production of lipids and renewal of the epidermis. It has been shown to replenish and nourish the skin, leaving it smoother and plumper; it also is nontoxic to humans, natural and environmentally friendly. The general range of use level is from $0.3-2.5\mathrm{\%}$$0.3-2.5\mathrm{\%}$0.3-2.5%0.3-2.5 \%, depending on the product form being deployed for end product use. The acute oral toxicity test for $\gamma$$\gamma$gamma\gamma-PGA in the form of its sodium salt gave an ${\mathrm{LD}}_{50}>5,000$${\mathrm{LD}}_{50}>5,000$LD_(50) > 5,000\mathrm{LD}_{50}>5,000 $\mathrm{mg}/\mathrm{kg}/$$\mathrm{mg}/\mathrm{kg}/$mg//kg//\mathrm{mg} / \mathrm{kg} / day. Biodegradability tests of the sodium salt of $\gamma$$\gamma$gamma\gamma-PGA revealed that $90\mathrm{\%}$$90\mathrm{\%}$90%90 \% degraded in 180 days. Table 1 compares characteristics of $\gamma$$\gamma$gamma\gamma-PGA to other peptides.
如圖 6 所示，含有 $\gamma$$\gamma$gamma\gamma -PGA 的產品可增強皮膚的支撐結構，為細胞提供能量，刺激脂質的產生和表皮的更新。它已被證明可以補充和滋養皮膚，使其更光滑、更豐滿;它還對人類無毒，自然且環保。一般使用級別範圍為 ， $0.3-2.5\mathrm{\%}$$0.3-2.5\mathrm{\%}$0.3-2.5%0.3-2.5 \% 具體取決於為最終產品使用而部署的產品形式。鈉鹽形式的 $\gamma$$\gamma$gamma\gamma -PGA 的急性口服毒性試驗給出了 ${\mathrm{LD}}_{50}>5,000$${\mathrm{LD}}_{50}>5,000$LD_(50) > 5,000\mathrm{LD}_{50}>5,000 $\mathrm{mg}/\mathrm{kg}/$$\mathrm{mg}/\mathrm{kg}/$mg//kg//\mathrm{mg} / \mathrm{kg} / 一天。 $\gamma$$\gamma$gamma\gamma -PGA 鈉鹽的生物降解性測試表明， $90\mathrm{\%}$$90\mathrm{\%}$90%90 \% 在180天內降解。表 1 比較了 $\gamma$$\gamma$gamma\gamma -PGA 與其他肽的特性。
$\gamma$$\gamma$gamma\gamma-PGA contains a robust amount of vitamins, oligiosaccharides and beneficial enzymes currently not found in the nonfermented soybean product. ${}^{12}$${}^{12}$^(12){ }^{12} It can be described as a white to off-white, granulated, free-flowing powder. Purity by HPLC is a minimum of $70\mathrm{\%}$$70\mathrm{\%}$70%70 \%, and the minimum assay value determined by gravimetric analysis of the salt of
$\gamma$$\gamma$gamma\gamma -PGA 含有大量維生素、寡糖和有益酶，目前在非發酵大豆產品中不存在。 ${}^{12}$${}^{12}$^(12){ }^{12} 它可以被描述為白色至灰白色、顆粒狀、自由流動的粉末。HPLC 的純度是 $70\mathrm{\%}$$70\mathrm{\%}$70%70 \% 的最小值，通過鹽的重量分析確定的最小測定值
the $\gamma$$\gamma$gamma\gamma-PGA is no less than $90\mathrm{\%}\mathrm{wt}/\mathrm{wt}$$90\mathrm{\%}\mathrm{wt}/\mathrm{wt}$90%wt//wt90 \% \mathrm{wt} / \mathrm{wt}. The loss on drying is no more than $10\mathrm{\%}\mathrm{wt}/\mathrm{wt}\gamma$$10\mathrm{\%}\mathrm{wt}/\mathrm{wt}\gamma$10%wt//wtgamma10 \% \mathrm{wt} / \mathrm{wt} \gamma-PGA. The heavy metals composition is no more than $0.002\mathrm{\%}$$0.002\mathrm{\%}$0.002%0.002 \% and the total bacterial count test for
- $\gamma$$\gamma$gamma\gamma PGA 不小於 $90\mathrm{\%}\mathrm{wt}/\mathrm{wt}$$90\mathrm{\%}\mathrm{wt}/\mathrm{wt}$90%wt//wt90 \% \mathrm{wt} / \mathrm{wt} 。乾燥損失不超過 $10\mathrm{\%}\mathrm{wt}/\mathrm{wt}\gamma$$10\mathrm{\%}\mathrm{wt}/\mathrm{wt}\gamma$10%wt//wtgamma10 \% \mathrm{wt} / \mathrm{wt} \gamma -PGA。重金屬成分不多 $0.002\mathrm{\%}$$0.002\mathrm{\%}$0.002%0.002 \% 於和總細菌計數測試
microorganisms is not greater than 500 CFU/g for a cosmetic-grade material. Salmonella and Escherichia coli are absent in $\gamma$$\gamma$gamma\gamma-PGA. Its particle size is $100\mathrm{\%}$$100\mathrm{\%}$100%100 \% through 100 mesh screen.
對於化妝品級材料，微生物不大於 500 CFU/g。-PGA 中 $\gamma$$\gamma$gamma\gamma 不存在沙門氏菌和大腸桿菌。其粒徑通過 $100\mathrm{\%}$$100\mathrm{\%}$100%100 \% 100目篩網。

Figure 6. Comparison of collagen, HA and $\gamma -$$\gamma -$gamma-\gamma--PGA in various parameters over a 28-day period
圖 6.28 天內各種參數的膠原蛋白、HA 和 $\gamma -$$\gamma -$gamma-\gamma- -PGA 的比較