Chen Daoyou, Rong Mingli, Tang Shuting, Zhang Chuanxi, Wei Hao, Yuan Zhaoting, Miao Tingwei, Song Hucheng, Jiang Haiming, Yang Yan, Zhang Lujia
Shanghai Engineering Research Center of Molecular Therapeutics & New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200062, China.
School of Food Science and Technology, Shihezi University, Shihezi 832000, China.
Food Chem. 2025 Mar 15;468:142385. doi: 10.1016/j.foodchem.2024.142385. Epub 2024 Dec 5.
This study aimed to explore the effect of directed enzymolysis on the umami characteristics of S. rugosoannulata, clarify the flavour formation mechanism of umami peptides. We expressed a new aminopeptidase (DNPEP) and obtained the umami peptides of S. rugosoannulata by alkaline protease and DNPEP. The optimal enzymolysis conditions were temperature 55 °C, solid-liquid ratio 1:20 (g/mL), alkaline protease enzymolysis (60 min, 0.5 %, pH 9.0), and DNPEP enzymolysis (80 min, 0.3 %, pH 8.0). The umami peptide components were separated by ultrafiltration and gel filtration chromatography. Six umami peptides (EEAKFN, KAELDLH, LADVEEDK, LKEAHDVA, AHLDYGDGK, and LGKSEDDVSK) were identified by LC-MS/MS and virtual screening, and the umami thresholds of the peptides were 0.15-1.09 mmol/L. Molecular simulations revealed that the amino acid residues Glu301, Ser217, Asp218, and Arg277 were crucial in the binding of the umami peptide to the T1R1/T1R3. Therefore, this study provides a theoretical basis for the development of mushroom condiments.
本研究旨在探讨定向酶解对皱环盖菇鲜味特性的影响，阐明鲜味肽的风味形成机制。我们表达了一种新的氨肽酶（DNPEP），并通过碱性蛋白酶和DNPEP获得了皱环盖菇的鲜味肽。最佳酶解条件为温度55℃、固液比1:20（g/mL）、碱性蛋白酶酶解（60分钟，0.5%，pH 9.0）和DNPEP酶解（80分钟，0.3%，pH 8.0）。通过超滤和凝胶过滤色谱分离鲜味肽成分。通过LC-MS/MS和虚拟筛选鉴定出六种鲜味肽（EEAKFN、KAELDLH、LADVEEDK、LKEAHDVA、AHLDYGDGK和LGKSEDDVSK），这些肽的鲜味阈值为0.15 - 1.09 mmol/L。分子模拟表明，氨基酸残基Glu301、Ser217、Asp218和Arg277在鲜味肽与T1R1/T1R3的结合中起关键作用。因此，本研究为蘑菇调味品的开发提供了理论依据。
