Whitening and immunopotentiating activity of polysaccharide produced by marine bacteria

由海洋假单胞菌sp. WAK-1产生的多糖是由n -乙酰- d -半乳糖胺、n -乙酰- l-半乳糖胺、d -葡萄糖醛酸和丙酮酸按2:1:1:1的摩尔比组成的高分子量糖胺聚糖。多糖具有美白和免疫增强活性。本研究报道了海洋多糖抑制小鼠B-16黑色素瘤细胞系黑色素生物合成的机制，以及T辅助（Th） 1型细胞在小鼠J774.1巨噬细胞样细胞系和原代脾细胞中促进细胞介导的免疫反应。多糖对B-16细胞的黑色素合成有抑制作用，但以L-DOPA为底物的糖胺聚糖对蘑菇和B-16的酪氨酸酶无抑制作用。RT-PCR检测黑色素生物合成酶基因表达结果表明，多糖抑制酪氨酸酶的表达，但不抑制酪氨酸酶相关蛋白-1和-2的表达。多糖分别促进J774.1巨噬细胞样细胞和正常脾细胞产生白细胞介素(IL)-12和干扰素(IFN)-γ作为Th-1型细胞因子。此外，还建立了利用超滤膜柱纯化假单胞菌WAK-1培养滤液中糖胺聚糖的大雄性方法。本报告中所描述的多糖，我们也于2006年5月12日获得了增白剂专利（No.3802011），并于2006年10月6日获得了商标名（No.4993867） uminglycan ^<TM>。因此，该海洋糖胺聚糖可用于开发美白化妆品和保健食品。

The polysaccharide produced by a marine Pseudomonas sp. WAK-1 was found to be a high molecular-weigh glycosaminoglycan composed of N-acetyl-D-galactosamine, N-acetyl-L-galactosamine, D-glucuronic acid and pyruvate in a molar ratio of 2:1:1:1. The polysaccharide has whitening and immunopotentiating activity. In this research, we report the mechanism of the inhibition of melanin biosynthesis in mouse B-16 melanoma cell line and the cell-mediated immune response promoted by T helper (Th) 1 type cells in mouse J774.1 macrophage-like cell line and primary spleen cells by the marine polysaccharide. The polysaccharide suppressed melanin biosynthesis in B-16 cells without cytotoxicity However using L-DOPA as the substrate no inhibitory activity of tyrosinases from mushroom and B-16 coils by the glycosaminoglycan was observed. RT-PCR for gene expression of enzymes in melanin biosynthesis demonstrated that the polysaccharide inhibited the expression of tyrosinase, but not of tyrosinase related protein-1 and -2. The polysaccharide enhanced productions of interleukin (IL)-12 and interferon (IFN)-γ as Th-1 type cytokines by J774.1 macrophage-like cells and normal splenocytes respectively In addition, large male purification method of the glycosaminoglycan from culture filtrate of Pseudomonas sp. WAK-1 was also developed using column of ultrafiltration membrane. As for the polysaccharide descried in this repot, we also obtained a patent (No.3802011) as a whitening agent in May 12, 2006 and a brand name (No.4993867) as Uminoglycan^<TM> in October 6, 2006. Therefore, this marine glycosaminoglycan can be useful for development of whitening cosmetics and health foods.