Whitening and immunopotentiating activity of polysaccharide produced by marine bacteria

海洋假单胞菌产生的多糖。WAK-1是一种由N-乙酰-D-氨基半乳糖、N-乙酰-L-氨基半乳糖、D-葡萄糖醛酸和丙酮酸组成的高相对分子质量糖胺聚糖，其摩尔比为2：1：1：1，具有美白和免疫增强活性。在本研究中，我们报道了海洋多糖抑制小鼠B-16黑色素瘤细胞系黑素生物合成的机制，以及海洋多糖对小鼠J774.1巨噬细胞样细胞系和原代脾细胞的T辅助细胞(Th)1型细胞免疫应答的促进作用。该多糖对B-16细胞黑色素的生物合成无细胞毒性，但以L多巴为底物，其对蘑菇酪氨酸酶和B-16盘状酪氨酸酶的抑制作用不明显。黑素生物合成相关酶基因表达的RT-PCR结果显示，该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。该多糖可促进J774.1巨噬细胞样细胞和正常脾细胞产生Th-1型细胞因子IL-12和干扰素-γ。此外，还介绍了从假单胞菌培养滤液中分离纯化糖胺多糖的方法。WAK-1也是利用超滤膜柱研制而成。对于本报告中描述的多糖，我们还于2006年5月12日获得了美白剂专利(编号3802011)，并于2006年10月6日获得了Uminoglycan^&lt；TM&gt；的商标名称(编号：4993867)。因此，这种海洋糖胺聚糖可用于美白化妆品和保健食品的开发。

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.