Peng Pai, Yu Hui, Xian Meiting, Qu Caiye, Guo Zhiqiang, Li Shuyi, Zhu Zhenzhou, Xiao Juan
Hainan Engineering Research Center of Aquatic Resources Efficient Utilization in South China Sea, Key Laboratory of Seafood Processing of Haikou, School of Food Science and Engineering, Hainan University, Haikou 570228, China.
School of Marine Science and Engineering, Hainan University, Haikou 570228, China.
Mar Drugs. 2025 Feb 9;23(2):75. doi: 10.3390/md23020075.
Bioactive peptides represent a promising therapeutic approach for Alzheimer's disease (AD) by maintaining cholinergic system homeostasis through the inhibition of acetylcholinesterase (AChE) activity. This study focused on extracting AChE inhibitory peptides from yellowfin tuna pancreas using moderate ultrasound-assisted enzymatic hydrolysis (MUE). Firstly, papain and MUE stood out from five enzymes and four enzymatic hydrolysis methods, respectively, by comparing the degree of hydrolysis and AChE inhibitory activity of different pancreatic protein hydrolysates. Subsequently, the optimal MUE conditions were obtained by single-factor, Plackett-Burman, and response surface methodologies. The pancreatic protein hydrolysate prepared under optimal MUE conditions was then purified by ultrafiltration followed by RP-HPLC, from which a novel AChE inhibitory peptide (LLDF) was identified by LC-MS/MS and virtual screening. LLDF effectively inhibited AChE activity by a competitive inhibition mechanism, with an IC of 18.44 ± 0.24 μM. Molecular docking and molecular dynamic simulation revealed that LLDF bound robustly to the active site of AChE via hydrogen bonds. These findings provided a theoretical basis for the valuable use of yellowfin tuna pancreas and introduced a new viewpoint on the potential therapeutic advantages of AChE inhibitory peptides for future AD treatment.
生物活性肽通过抑制乙酰胆碱酯酶（AChE）活性来维持胆碱能系统的稳态，是一种很有前景的阿尔茨海默病（AD）治疗方法。本研究聚焦于利用适度超声辅助酶解（MUE）从黄鳍金枪鱼胰腺中提取AChE抑制肽。首先，通过比较不同胰腺蛋白水解物的水解程度和AChE抑制活性，木瓜蛋白酶和MUE分别在五种酶和四种酶解方法中脱颖而出。随后，通过单因素法、Plackett-Burman法和响应面法获得了最佳的MUE条件。然后，在最佳MUE条件下制备的胰腺蛋白水解物通过超滤继以反相高效液相色谱（RP-HPLC）进行纯化，通过液相色谱-串联质谱（LC-MS/MS）和虚拟筛选从中鉴定出一种新型AChE抑制肽（LLDF）。LLDF通过竞争性抑制机制有效抑制AChE活性，其半数抑制浓度（IC）为18.44±0.24μM。分子对接和分子动力学模拟表明，LLDF通过氢键与AChE的活性位点紧密结合。这些发现为黄鳍金枪鱼胰腺的宝贵利用提供了理论依据，并为AChE抑制肽在未来AD治疗中的潜在治疗优势引入了新的观点。
