地龙胰脂肪酶抑制肽的制备、结构鉴定及活性分析OA

The advantages of endogenous enzymes and commercial enzymes were combined to obtain earthworm protein peptides with efficient pancreatic lipase inhibition activity using a two-step enzymatic hydrolysis method to hydrolyze earthworm protein.The pancreatic lipase inhibition rate was used as the evaluation index,and single-factor experiments and orthogonal experiments were used to optimize the second-step enzymatic hydrolysis process.In addition,the hydrolysates were purified using ultrafiltration and Sephadex G-15 gel chromatography,and peptide segments with high pancreatic lipase inhibitory activity were screened by combining LC-MS/MS,molecular docking,and other technologies with the PeptideRanker database.The results showed that the optimal preparation process for the second-step enzymatic hydrolysis involved a solid:liquid ratio of 1:2(50%,m/m),alkaline protease dosage of 6.0×104 U,temperature of 50℃,pH value of 9.5,and enzymatic hydrolysis time of 3 hours.Under these conditions,the pancreatic lipase inhibition activity reached 53.92%,the hydrolysis degree was 41.98%,and the soluble peptide yield reached 89.17%.A total of 10 peptide segments with high affinity,good biological activity,and non-toxicity were screened,namely GDAPPFFP,YPFDPGP,WGPDLP,FDFP,DDFFR,FLDF,SDDGFF,FDDF,SDDFF,and WDDFF.Small molecule peptides and amino acids are thought to be the primary substances causing the inhibitory effect of earthworm protein on pancreatic lipase.Overall,this study provides a new perspective for understanding the lipid-lowering activity of earthworm protein.It establishes a foundation for further research on the structure-activity relationship of earthworm protein functional peptides and the mechanisms of pancreatic lipase inhibition.