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Enrichment of Extracellular Matrix Proteins from Tissues and Digestion into Peptides for Mass Spectrometry Analysis
Published on: July 23, 2015
Low-Molecular-Weight Salmon Collagen Peptides Promote Extracellular Matrix Gene (ECM) Expression in BJ Fibroblasts
Soottawat Benjakul1, Krisana Nilsuwan1, Umesh Patil1
1International Center of Excellence in Seafood Science and Innovation, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai 90110, Songkhla, Thailand.
Abstract:
Salmon skin, a high-volume by-product of seafood processing, offers a circular-bioeconomy route to sustainable, value-added ingredients. This study aimed to generate low-molecular-weight collagen peptides (LMWCPs) from salmon skin using a stepwise enzymatic process and to evaluate their safety and pro-extracellular-matrix (ECM) activity in human BJ fibroblasts. LMWCPs were produced by sequential hydrolysis (alcalase/papain, then collagenase) and characterized as low-molecular-weight peptide preparations with a mean dispersed particle diameter of approximately 117 nm. LMWCPs display negatively charged peptide dispersions with a mass centered around ~1 kDa. Cytocompatibility (MTT) showed no toxicity up to 1.5 mg/mL over 48 h. Gene expression by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) revealed a robust, dose-dependent ECM response: collagen type I alpha 1 chain (COL1A1) increased by approximately 7-fold, with additional rises of 5-6-fold in versican (VCAN) levels and modest increases in elastin (ELN) and transforming growth factor-β (TGF-β). These findings provide an exploratory process-to-phenotype link between the two-step hydrolysis process, physicochemical characteristics of the resulting LMWCPs, and changes in ECM-related gene expression in BJ fibroblasts. Overall, this study demonstrates that salmon skin LMWCPs were cytocompatible within the tested concentration range and modulated several ECM-related genes, providing a preliminary basis for future protein-level, functional, and translational evaluation.
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