Chum salmon myofibrillar protein (Mf) digested by pepsin-trypsin (dMf) was conjugated to alginate oligosaccharide (AO) through the Maillard reaction between the lysine residues in the dMf and the reducing end-groups of AO. The dMf-AO conjugate strongly suppressed the production of inflammatory cytokines, particularly tumor necrosis factor-α and interleukin-6, in lipopolysaccharide-stimulated murine macrophages. The anti-inflammatory effect was enhanced with increasing amounts of AO bound to dMf through the Maillard reaction, and the subsequent process of the Maillard reaction was involved in the enhancement of the anti-inflammatory activity. AO-conjugated peptides, the major anti-inflammatory components contained in the dMf-AO conjugate, were successfully recovered using isoelectric focusing without the use of carrier ampholytes (autofocusing); they were effectively concentrated in acidic peptide fractions, with the negative charge of the carboxyl groups of AO contributing to this separation. In conclusion, the combined use of AO conjugation and autofocusing is a useful method for producing novel anti-inflammatory peptides from fish Mf.

通过胃蛋白酶-胰蛋白酶（dMf）消化的鲑鱼肌原纤维蛋白（Mf）通过dMf中赖氨酸残基与AO还原端基之间的美拉德反应与藻酸盐寡糖（AO）结合。dMf-AO共轭物在脂多糖刺激的鼠巨噬细胞中强烈抑制了炎性细胞因子的产生，特别是肿瘤坏死因子-α和白介素-6的产生。通过美拉德反应，与dMf结合的AO量增加，抗炎作用增强，而美拉德反应的后续过程参与了抗炎活性的增强。dMf-AO缀合物中主要的抗炎成分是AO缀合肽，它是通过等电聚焦成功回收的，而无需使用载体两性电解质（自动聚焦）。它们有效地浓缩在酸性肽馏分中，而AO羧基的负电荷有助于这种分离。总之，结合使用AO结合和自动聚焦是从鱼类Mf生产新型抗炎肽的有用方法。