A Hybrid Peptide DEFB-TP5 Expressed in Methylotrophic Yeast Neutralizes LPS With Potent Anti-inflammatory Activities

Abstract

DEFB-TP5 is a novel auspicious health-beneficial peptide derivative from two naturally occurring peptides, b-Defensin (DEFB) and thymopentin (TP5), and shows strong antiinflammatory activity and binds to LPS without cytotoxicity and hemolytic effect. Furthermore, the application of DEFB-TP5 peptide is inadequate by its high cost. In the current study, we developed a biocompatible mechanism for expression of the DEFB-TP5 peptide in Pichia pastoris. The transgenic strain of hybrid DEFB-TP5 peptide with a molecular weight of 6.7kDa as predictable was obtained. The recombinant DEFB-TP5 peptide was purified by Ni-NTA chromatography, estimated 30.41 mg/L was obtained from the cell culture medium with 98.2% purity. Additionally, The purified DEFB-TP5 peptide significantly (p< 0.05) diminished the release of nitric oxide (NO), TNF-a, IL-6, IL-1b in LPS-stimulated RAW264.7 macrophages in a dose-dependent manner. This study will not only help to understand the molecular mechanism of expression that can potentially be used to develop an anti-endotoxin peptide but also to serve as the basis for the development of antimicrobial and anti-inflammatory agents as well, which also provides a potential source for the production of recombinant bioactive DEFB-TP5 at the industrial level.

Yeast Expressed Hybrid Peptide CLP Abridged Pro-Inflammatory Cytokine Levels by Endotoxin Neutralization

“…In this study, the LAL test demonstrated that CLP neutralized its endotoxin activity, thereby blocking the endotoxin-induced activation of downstream signaling pathways. Furthermore, the effect on LPS inhibition was better than the previously expressed peptides DEFB-TP5 [ 12 ] and CATH-2TP5 [ 16 ].…”

“…AMPs can disrupt bacterial cell membranes through the carpet, toroidal-pore, and barrel-stave disruption mechanisms, which is quite different from conventional drugs [ 7 ]. In addition to antibacterial activities, many AMPs also have significant anti-inflammatory activities [ 8 , 9 , 10 , 11 , 12 ]. This is because some AMPs can directly interact with LPS, thereby effectively alleviating LPS-induced inflammation [ 13 , 14 , 15 ].…”