Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic Hydrolysis

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 modera...

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Main Authors: Pai Peng, Hui Yu, Meiting Xian, Caiye Qu, Zhiqiang Guo, Shuyi Li, Zhenzhou Zhu, Juan Xiao
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Marine Drugs
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Online Access:https://www.mdpi.com/1660-3397/23/2/75
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author Pai Peng
Hui Yu
Meiting Xian
Caiye Qu
Zhiqiang Guo
Shuyi Li
Zhenzhou Zhu
Juan Xiao
author_facet Pai Peng
Hui Yu
Meiting Xian
Caiye Qu
Zhiqiang Guo
Shuyi Li
Zhenzhou Zhu
Juan Xiao
author_sort Pai Peng
collection DOAJ
description 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<sub>50</sub> 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.
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spelling doaj-art-4befa9f85b36495cb4592891a789cfb72025-08-20T02:03:28ZengMDPI AGMarine Drugs1660-33972025-02-012327510.3390/md23020075Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic HydrolysisPai Peng0Hui Yu1Meiting Xian2Caiye Qu3Zhiqiang Guo4Shuyi Li5Zhenzhou Zhu6Juan Xiao7Hainan 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, ChinaHainan 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, ChinaHainan 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, ChinaHainan 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, ChinaSchool of Marine Science and Engineering, Hainan University, Haikou 570228, ChinaNational R&D Center for Se-Rich Agricultural Products, Processing, Hubei Engineering Research Center for Deep Processing of Green, Se-Rich Agricultural Products, School of Modern Industry for Selenium Science and Engineering, Wuhan Polytechnic University, Wuhan 430023, ChinaNational R&D Center for Se-Rich Agricultural Products, Processing, Hubei Engineering Research Center for Deep Processing of Green, Se-Rich Agricultural Products, School of Modern Industry for Selenium Science and Engineering, Wuhan Polytechnic University, Wuhan 430023, ChinaHainan 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, ChinaBioactive 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<sub>50</sub> 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.https://www.mdpi.com/1660-3397/23/2/75yellowfin tuna pancreasAlzheimer’s diseaseAChE inhibitory peptidesmoderate ultrasound-assisted enzymatic hydrolysispeptide-enzyme interaction
spellingShingle Pai Peng
Hui Yu
Meiting Xian
Caiye Qu
Zhiqiang Guo
Shuyi Li
Zhenzhou Zhu
Juan Xiao
Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic Hydrolysis
Marine Drugs
yellowfin tuna pancreas
Alzheimer’s disease
AChE inhibitory peptides
moderate ultrasound-assisted enzymatic hydrolysis
peptide-enzyme interaction
title Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic Hydrolysis
title_full Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic Hydrolysis
title_fullStr Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic Hydrolysis
title_full_unstemmed Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic Hydrolysis
title_short Preparation of Acetylcholinesterase Inhibitory Peptides from Yellowfin Tuna Pancreas Using Moderate Ultrasound-Assisted Enzymatic Hydrolysis
title_sort preparation of acetylcholinesterase inhibitory peptides from yellowfin tuna pancreas using moderate ultrasound assisted enzymatic hydrolysis
topic yellowfin tuna pancreas
Alzheimer’s disease
AChE inhibitory peptides
moderate ultrasound-assisted enzymatic hydrolysis
peptide-enzyme interaction
url https://www.mdpi.com/1660-3397/23/2/75
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