Chen GuoXing, Ge Xiaofan, Sun Yuting, Sui Wenjie, Jin Yan, Geng Jieting, Zhang Min, Wu Tao
State Key Laboratory of Food Nutrition and Safety, Engineering Research Center of Food Biotechnology, Ministry of Education, Tianjin University of Science & Technology, Tianjin 300457, China.
Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan.
Int J Biol Macromol. 2025 Mar;295:139499. doi: 10.1016/j.ijbiomac.2025.139499. Epub 2025 Jan 3.
This study aimed to identify novel α-amylase inhibitory peptides from Russian sea cucumbers and elucidate their inhibitory mechanisms. Among the 52 identified sea cucumber peptide (SCP), two peptides with potential α-amylase inhibitory activity, FPSPPLVA (SCP1) and GPPMPPPPLP (SCP2), were selected from the sequences researched. The results showed that both SCP1 and SCP2 exhibited α-amylase inhibitory activity with IC of 0.92 ± 0.03 mg/mL (SCP1) and 2.01 ± 0.02 mg/mL (SCP2), respectively. Molecular docking studies revealed the potential interaction mechanism between these inhibitors and α-amylase. Two peptides interacted with Gln63, Tyr151, Thr163, Lys200, His201, His305, and Ala307 within the α-amylase active site. The formation of hydrogen bonds and hydrophobic interactions between the inhibitors and α-amylase impedes the formation of enzyme-substrate complexes and subsequent glycosylation. Lineweaver-Burk plot indicated that both SCP1 and SCP2 act as reversible mixed inhibitors of α-amylase. Furthermore, the results of infrared and circular dichroism spectroscopy confirmed the formation of non-covalent binding complexes between SCP and amylase, leading to alterations in the secondary structure of the enzyme.
本研究旨在从俄罗斯海参中鉴定新型α-淀粉酶抑制肽，并阐明其抑制机制。在鉴定出的52种海参肽（SCP）中，从研究序列中选出了两种具有潜在α-淀粉酶抑制活性的肽，即FPSPPLVA（SCP1）和GPPMPPPPLP（SCP2）。结果表明，SCP1和SCP2均表现出α-淀粉酶抑制活性，其IC50分别为0.92±0.03mg/mL（SCP1）和2.01±0.02mg/mL（SCP2）。分子对接研究揭示了这些抑制剂与α-淀粉酶之间潜在的相互作用机制。两种肽与α-淀粉酶活性位点内的Gln63、Tyr151、Thr163、Lys200、His201、His305和Ala307相互作用。抑制剂与α-淀粉酶之间形成的氢键和疏水相互作用阻碍了酶-底物复合物的形成及随后的糖基化。Lineweaver-Burk图表明，SCP1和SCP2均作为α-淀粉酶的可逆混合型抑制剂起作用。此外，红外光谱和圆二色光谱的结果证实了SCP与淀粉酶之间形成了非共价结合复合物，导致酶的二级结构发生改变。
