Ninghai Institute of Mariculture Breeding and Seed Industry, Zhejiang Wanli University, Ningbo 315604, China; Zhejiang Key Laboratory of Intelligent Food Logistic and Processing, Zhejiang-Malaysia Joint Research Laboratory for Agricultural Product Processing and Nutrition, College of Food Science and Engineering, Ningbo University, Ningbo 315211, China.
Zhejiang Key Laboratory of Intelligent Food Logistic and Processing, Zhejiang-Malaysia Joint Research Laboratory for Agricultural Product Processing and Nutrition, College of Food Science and Engineering, Ningbo University, Ningbo 315211, China.
Food Res Int. 2024 Aug;189:114534. doi: 10.1016/j.foodres.2024.114534. Epub 2024 May 21.
In order to identify the peptides responsible for bitter defects and to understand the mechanism of bitterness in dry-cured ham, the peptides were identified by LC-MS/MS, and the interaction between bitter peptides and receptor proteins were evaluated by molecular docking and molecular dynamics simulation; the signal transduction mechanism of bitter peptides was investigated using the model of HEK-293T cells by calcium imaging and transcriptomics analysis. The results of LC-MS/MS showed that 11 peptides were identified from the high bitterness fraction of defective ham; peptides PKAPPAK, VTDTTR and YIIEK derived from titin showed the highest bitterness values compared with other peptides. The results of molecular docking showed that lower CDOCKER energy was observed in the interaction between these peptides and hT2R16 in comparison with these receptors of hT2R1, hT2R4, hT2R5, hT2R8 and hT2R14, and the interaction of hT2R16 and peptides was stabilized by hydrophobic interaction and hydrogen bond. The average RMSF values of VTDTTR were higher than that of YIIEK and PKAPPAK, while EC values of VTDTTR were lower compared with PKAPPAK and YIIEK. Transcriptomics analysis showed that 529 differentially expressed genes were identified in HEK-293T cells during the stimulating by VTDTTR and were mainly enriched into neuroactive ligand-receptor interaction, MAPK pathway, cAMP pathway and calcium signaling pathway, which were mainly responsible for the bitter signal transduction of VTDTTR. These results could provide evidence for understanding the bitter defects of dry-cured ham and the taste mechanism of bitter peptide.
为了鉴定导致干腌火腿苦味缺陷的肽，并了解苦味的机制，通过 LC-MS/MS 鉴定肽，通过分子对接和分子动力学模拟评估苦味肽与受体蛋白的相互作用；通过钙成像和转录组学分析，使用 HEK-293T 细胞模型研究苦味肽的信号转导机制。LC-MS/MS 的结果表明，从有缺陷的火腿的高苦味部分鉴定出 11 种肽；与其他肽相比，来自肌联蛋白的肽 PKAPPAK、VTDTTR 和 YIIEK 表现出最高的苦味值。分子对接的结果表明，与 hT2R1、hT2R4、hT2R5、hT2R8 和 hT2R14 这些受体相比，这些肽与 hT2R16 的相互作用观察到更低的 CDOCKER 能量，并且 hT2R16 与肽的相互作用通过疏水相互作用和氢键稳定。与 YIIEK 和 PKAPPAK 相比，VTDTTR 的平均 RMSF 值较高，而与 PKAPPAK 和 YIIEK 相比，VTDTTR 的 EC 值较低。转录组学分析表明，在 VTDTTR 刺激 HEK-293T 细胞期间，鉴定出 529 个差异表达基因，主要富集到神经活性配体-受体相互作用、MAPK 途径、cAMP 途径和钙信号转导途径，主要负责 VTDTTR 的苦味信号转导。这些结果可为理解干腌火腿的苦味缺陷和苦味肽的味道机制提供证据。
