Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed Peptides
In this study, we systematically investigated the regulatory effects of two hempseed peptides (APAM and RLPA) on pancreatic lipase (PL) and cholesterol esterase (CE) activities and the molecular binding mechanism using enzyme kinetics, synergistic inhibition, fluorescence spectroscopy, isothermal ti...
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China Food Publishing Company
2025-01-01
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| Online Access: | https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-2-005.pdf |
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| author | YIN Hao, ZHU Jiangxiong, ZHAO Haiyun, ZHONG Yu, WANG Danfeng, DENG Yun |
| author_facet | YIN Hao, ZHU Jiangxiong, ZHAO Haiyun, ZHONG Yu, WANG Danfeng, DENG Yun |
| author_sort | YIN Hao, ZHU Jiangxiong, ZHAO Haiyun, ZHONG Yu, WANG Danfeng, DENG Yun |
| collection | DOAJ |
| description | In this study, we systematically investigated the regulatory effects of two hempseed peptides (APAM and RLPA) on pancreatic lipase (PL) and cholesterol esterase (CE) activities and the molecular binding mechanism using enzyme kinetics, synergistic inhibition, fluorescence spectroscopy, isothermal titration calorimetry and molecular docking. The results showed that RLPA had stronger inhibitory activity on PL and CE with half maximal inhibitory concentration (IC50) values of (79.62 ± 3.20) and (301.27 ± 14.40) μmol/L, respectively. Kinetic analysis indicated that the inhibition mechanism of APAM on the two enzymes was competitive inhibition, whereas RLPA was a mixed-type inhibitor. Both APAM and RLPA synergistically inhibited PL and CE with orlistat at low concentrations, but the synergistic effect was weakened or even antagonized at high concentrations. Fluorescence spectroscopy results showed that both peptides could produce static quenching by binding to PL and CE, which changed the hydrophobic environment of aliphatic amino acids in the enzymes. Isothermal titration calorimetry demonstrated that the binding process between the peptides and the enzymes was spontaneous and exothermic, which was achieved mainly through hydrogen bonding and electrostatic interaction. Further analysis using molecular docking simulations showed that hydrogen bonding, salt bridges, and cation-π interactions played crucial roles in peptide-enzyme binding. This study enriches our understanding of the interaction mechanism of peptides with PL and CE, and provides a scientific basis for the development of functional foods based on hempseed protein. |
| format | Article |
| id | doaj-art-a8f8a475910c4a9f9b7bbb509d5ef9ce |
| institution | Kabale University |
| issn | 1002-6630 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | China Food Publishing Company |
| record_format | Article |
| series | Shipin Kexue |
| spelling | doaj-art-a8f8a475910c4a9f9b7bbb509d5ef9ce2025-02-05T09:08:22ZengChina Food Publishing CompanyShipin Kexue1002-66302025-01-01462384910.7506/spkx1002-6630-20240705-061Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed PeptidesYIN Hao, ZHU Jiangxiong, ZHAO Haiyun, ZHONG Yu, WANG Danfeng, DENG Yun0(1. School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China;2. Yunnan Dali Research Institute of Shanghai Jiao Tong University, Dali 671000, China;3. Yunnan Dali Bai Autonomous Prefecture Inspection and Testing Institute, Dali 671000, China)In this study, we systematically investigated the regulatory effects of two hempseed peptides (APAM and RLPA) on pancreatic lipase (PL) and cholesterol esterase (CE) activities and the molecular binding mechanism using enzyme kinetics, synergistic inhibition, fluorescence spectroscopy, isothermal titration calorimetry and molecular docking. The results showed that RLPA had stronger inhibitory activity on PL and CE with half maximal inhibitory concentration (IC50) values of (79.62 ± 3.20) and (301.27 ± 14.40) μmol/L, respectively. Kinetic analysis indicated that the inhibition mechanism of APAM on the two enzymes was competitive inhibition, whereas RLPA was a mixed-type inhibitor. Both APAM and RLPA synergistically inhibited PL and CE with orlistat at low concentrations, but the synergistic effect was weakened or even antagonized at high concentrations. Fluorescence spectroscopy results showed that both peptides could produce static quenching by binding to PL and CE, which changed the hydrophobic environment of aliphatic amino acids in the enzymes. Isothermal titration calorimetry demonstrated that the binding process between the peptides and the enzymes was spontaneous and exothermic, which was achieved mainly through hydrogen bonding and electrostatic interaction. Further analysis using molecular docking simulations showed that hydrogen bonding, salt bridges, and cation-π interactions played crucial roles in peptide-enzyme binding. This study enriches our understanding of the interaction mechanism of peptides with PL and CE, and provides a scientific basis for the development of functional foods based on hempseed protein.https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-2-005.pdflipid-digesting enzymes; inhibition mechanism; fluorescence spectroscopy; isothermal titration calorimetry; molecular modeling |
| spellingShingle | YIN Hao, ZHU Jiangxiong, ZHAO Haiyun, ZHONG Yu, WANG Danfeng, DENG Yun Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed Peptides Shipin Kexue lipid-digesting enzymes; inhibition mechanism; fluorescence spectroscopy; isothermal titration calorimetry; molecular modeling |
| title | Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed Peptides |
| title_full | Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed Peptides |
| title_fullStr | Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed Peptides |
| title_full_unstemmed | Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed Peptides |
| title_short | Molecular Mechanism of Targeted Inhibition of Pancreatic Lipase and Cholesterol Esterase by Hempseed Peptides |
| title_sort | molecular mechanism of targeted inhibition of pancreatic lipase and cholesterol esterase by hempseed peptides |
| topic | lipid-digesting enzymes; inhibition mechanism; fluorescence spectroscopy; isothermal titration calorimetry; molecular modeling |
| url | https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-2-005.pdf |
| work_keys_str_mv | AT yinhaozhujiangxiongzhaohaiyunzhongyuwangdanfengdengyun molecularmechanismoftargetedinhibitionofpancreaticlipaseandcholesterolesterasebyhempseedpeptides |