Zefen Wang,Siyuan Liu,Qian Zhou,Pengru Liu,Dankui Liao,Ping Lan,Bo Peng,Xiongdiao Lan
Immobilized angiotensin converting enzyme (ACE) utilizes specific interaction between ACE and ACE inhibitors to screen drugs for hypertension therapy, but traditional immobilized ACE is insufficient to achieve satisfactory effect for the poor immobilization efficiency and stability. In this work, mesoporous ZIF-8 (mZIF-8) coated with polydopamine (PDA) was developed as an enzyme-immobilization platform for the first time. Molecular docking was used to study the mechanism of ACE induced mZIF-8 synthesis. The immobilized enzyme ACE@mZIF-8/PDA were shown to possess mesoporous structures with ACE encapsulation efficiency of ∼90%. Compared with the free enzyme, ACE@mZIF-8/PDA exhibited enhanced tolerance to temperature, pH, unfolding and proteolytic agents, with excellent reusability (maintained 82.29% ± 4.11% of initial activity after being reused 10 times) and storage stability (retaining 93.12% ± 3.271% residual activity after 30 days). Molecular docking research conformed ACE induced the triggered nucleation of MOFs via binding with MOFs precursors. Moreover, ACE@mZIF-8/PDA was used as high-performance adsorbent for enrichment of ACE inhibitory peptides. The noncompetitive inhibitory peptide GF-6 and competitive inhibitory peptide VPP could be captured synchronously, which avoided the loss of active substances. These results indicated that the ACE@mZIF-8/PDA provides an evolutionary platform for rapid screening of inhibitory peptides.
Fangchao Cui,Liqing Xi,Guiqin Zhao,Dangfeng Wang,Xiqian Tan,Jianrong Li,Tingting Li
Gout, as the second major metabolic disease after cardiovascular disease. Peptides have the advantages of natural and no side effects in the treatment and prevention of gout. This study aimed to screen xanthine oxidase (XOD) inhibitory peptides from Scophthalmus maximus protein as a drug basis for the treatment of gout. The specific results are as follows: Immobilization of xanthine oxidase 25.564 μg/mg by modified magnetic beads, and modified magnetic beads can be reused three times. Three decapeptides (FSLVHYAGTV, FTNEKLQQFF and WDDMEKIWHH) were screened, the IC 50 were 3.741, 4.046 and 2.203 mg/mL. Circular dichroism and three-dimensional fluorescence spectra showed that the inhibitory peptide would change the helical structure of XOD and the internal hydrophobic environment. Molecular docking showed that the inhibitory effect of inhibitory peptides depends on the number of unique amino acid residues in the active center of XOD enzyme . In conclusion, this study provides a method for screening anti-gout peptides, which not only solves the problem of treating gout, but also realizes the high-value utilization of marine products. In addition, this method has substantial application value because of its low cost and simple operation.
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