Miso, a fermented paste made from soybeans, is used traditionally for seasoning of food. It has been a protein and nitrogen source since ancient times in Japan because of its high nutritional value. Furthermore, it has important health functions such as the estrogen-like activity of isoflavones, anti-oxidation, and angiotensin-converting-enzyme inhibition activity. Moreover, it has activity for neutralization of lipopolysaccharide (LPS) from Escherichia coli. Nevertheless, the mechanisms of that activity remain unclear. For this study, we purified and identified the proteins responsible for LPS-neutralization. After proteins were isolated from a miso extract using Blue native polyacrylamide gel electrophoresis, a protein found at 10 to 30 kDa on the polyacrylamide gel was identified using nano LC-MS/MS as 2S albumin in soybean (Glycine max). The protein had two LPS binding motifs: SKWQHQ (22 amino acid residues) and EKQKKKMEKE (131 amino acid residues). The protein in miso was found to have LPS neutralization activity, as assayed by prostaglandin D2 (PGD2 ) production from macrophage cells. The PGD2 production by macrophage cells was inhibited by LPS-neutralizing protein (LNP) from miso. Particularly, 50 mg/mL of LNP solution and LPS (10 µg/mL) inhibited production of PGD2 from the cells. The data were inferred as significantly different (P < 0.05) from statistical analyses by analysis of variance testing and Tukey tests. The 2S albumin in soybean is LNP, an LPS-neutralizing protein, produced in miso. PRACTICAL APPLICATION: A protein from miso fermented soy paste neutralizes an Escherichia coli intestinal bacterial product, lipopolysaccharide (LPS), which causes intestinal inflammation. Miso and its protein are anticipated for use as a probiotic agent to prevent intestinal inflammation in humans and domestic animals. Miso is useful not only as a seasoning for food, but also as a health-functional food because it is an LPS-neutralizing agent.

中文翻译：

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日本发酵豆酱味噌中的脂多糖中和蛋白

味噌是一种由大豆制成的发酵酱，传统上用于调味食物。由于营养价值高，日本自古以来就是蛋白质和氮的来源。此外，它还具有重要的保健功能，如异黄酮的雌激素样活性、抗氧化和血管紧张素转换酶抑制活性。此外，它还具有中和来自大肠杆菌的脂多糖 (LPS) 的活性。然而，该活动的机制仍不清楚。在这项研究中，我们纯化并鉴定了负责 LPS 中和的蛋白质。使用 Blue 天然聚丙烯酰胺凝胶电泳从味噌提取物中分离蛋白质后，使用纳米 LC-MS/MS 将聚丙烯酰胺凝胶上 10 至 30 kDa 的蛋白质鉴定为大豆中的 2S 白蛋白 (Glycine max)。该蛋白质具有两个 LPS 结合基序：SKWQHQ（22 个氨基酸残基）和 EKQKKKMEKE（131 个氨基酸残基）。发现味噌中的蛋白质具有 LPS 中和活性，正如通过巨噬细胞产生的前列腺素 D2 (PGD2) 所测定的。来自味噌的 LPS 中和蛋白 (LNP) 抑制巨噬细胞产生 PGD2。特别是，50 mg/mL 的 LNP 溶液和 LPS (10 µg/mL) 抑制了细胞中 PGD2 的产生。通过方差检验和 Tukey 检验分析，数据被推断为与统计分析显着不同 (P < 0.05)。大豆中的 2S 白蛋白是 LNP，一种 LPS 中和蛋白，在味噌中产生。实际应用：来自味噌发酵豆酱的蛋白质中和大肠杆菌肠道细菌产物脂多糖 (LPS)，从而引起肠道炎症。味噌及其蛋白质有望用作益生菌剂以预防人类和家畜的肠道炎症。味噌不仅可用作食品调味品，而且还可用作保健功能食品，因为它是一种 LPS 中和剂。