Rational sequence optimization is essential for transforming food-derived peptides into more potent angiotensin-converting enzyme (ACE) inhibitors. LYPVK and its sequence-designed derivative YWLKP exhibit potent activity; however, their binding mechanisms, transport routes, and in vivo antihypertensive effects remain unclear. Here, they were systematically investigated by biotechnological preparation, biolayer interferometry (BLI), molecular dynamics (MD) simulation, Caco-2 transport assays, and studies on spontaneously hypertensive rats (SHRs). BLI revealed that YWLKP bound to ACE with a subnanomolar affinity constant KD of (1.08 ± 0.40) × 10-9 M, consistent with MD results showing a more stable peptide-ACE complex. Caco-2 monolayer transport showed that LYPVK mainly permeated via the paracellular pathway, whereas YWLKP exhibited PepT1-mediated transport. In SHRs, both peptides significantly reduced blood pressure after single and long-term administration, accompanied by renin-angiotensin system modulation, reduced renal and cardiac impairment, and improved endothelial dysfunction. Collectively, these findings support YWLKP as a promising antihypertensive candidate for functional food applications.
Qingping Liang, Nana Sun, Changliang Zhu et al.· Journal of Agricultural and...· 0 citations
Recombinant humanized type III collagen has attracted increasing interest for biomedical and tissue engineering applications due to its roles in extracellular matrix remodeling and tissue repair. However, the production of long-continuous, structurally stable type III collagen fragment in Pichia pastoris is limited by host-derived proteolysis of the recombinant α1 chain, resulting in fragmentation and loss of structural integrity. In this study, a long-continuous type III collagen fragment was produced in P. pastoris by proteolytic selection and identified as a continuous 585-amino acid sequence (N611-P1195) by LC-MS/MS. Secretion efficiency was improved by signal peptide optimization, enabling the development of high-producing strains (6.22 g/L). rColIII was purified to > 90% purity with low endotoxin levels (< 10 EU/mg) using multimodal chromatography and exhibited enhanced thermal stability. Although CD analysis indicated the absence of a canonical triple-helix conformation, FITR spectroscopy confirmed preservation of key peptide backbone structures. Biological assays demonstrated favorable cytocompatibility, with rColIII promoting HSF cell viability, adhesion, and migration. This study provides a scalable strategy for producing long, continuous type III collagen fragment with improved stability in P. pastoris, and supports their potential applications in cosmetic, tissue engineering, regenerative medicine, and other biomedical fields.
Lina Zhu, Qingping Liang, Guijun Zhang et al.· Biotechnology and Bioenginee...· 0 citations
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