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To clarify the key flavor peptides that account for the cooked taste of puffer fish, this study was performed to examine flavor peptides extracted from the flesh of puffer fish (Takifugu obscurus).Peptides fractions (P1, P2, P3, P4 and P5) were purified from an aqueous extract of Takifugu obscurus muscle by ultrafiltration and Sephadex G-15 gel filtration chromatography (GFC).P2 was further fractionated into P2a,P2b, and P2c by reverse phase high performance liquid chromatography (RP-HPLC).Fraction P2b elicited umami and sweet taste.The amino acid sequence of P2b subfraction was identified as Tyr-Gly-Gly-Thr-Pro-Pro-Phe-Val (836.4 Da) by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/TOF MS/MS).Hydrophilic amino acids residues Tyr, Gly, Gly, Thr, and Phe are likely to contribute to the umami and sweet taste of this oetapeptide.The results of this study suggest this peptide is one of important components of the mellowness and tenderness taste of the Takifugu obscurus.