Seok-Chun Ko has completed his PhD from Jeju National University and Post-doctoral studies from Pukyong National University School of Biomedical Enginnering. He is the Research Professor of marine-integrated bionics Center. He has published more than 35 papers in SCI journals and has been serving as an Editorial Board Member of repute.
To prepare antioxidative peptide from Styela plicata, nine proteases were employed for enzymatic hydrolysis, and the antioxidative activities of the hydrolysates were investigated using electron spin resonance (ESR) spectrometer. Among the hydrolysates, tryptic hydrolysate exhibited the highest antioxidative activities than those of other enzymatic hydrolysates. In order to purity a peptide having potent antioxidative properties, tryptic hydrolysate was separated using consecutive chromatographic methods, and andtioxidative peptide was identifi ed to be Leu-Pro-His-Pro-Ser-Phe (696.3 Da) by Q-TOF ESI mass spectroscopy. It scavenged peroxyl, hydrolxyl and DPPH radicals at the IC50 values of 0.05, 1.98 and 0.17 mM, respectively. Pretreatment with the purifi ed peptide decreased the death of AAPH-treated cells, and reduced the generation of intracellular reactive oxygen species (ROS) in a dose-dependent manner in AAPH-treated cells. Furthermore, the purifi ed peptide signifi cantly reduced ROS generation and cell death in zebrafi sh model. Th ese results indicate that enzymatic hydrolysates of S. plicata protein possess potent antioxidative activity.
Van-Tinh Nguyen received his PhD from the Department of Biomedical Engineering, Pukyong National University, Korea. He graduated in 2012 from the University of Chosun and his current research interests include the isolation, safety and bioavailability of bioactive materials; development of marine-integrated cells and tissue regenerative biomedical substances.
This study examined the eff ects of Ciona intestinalis calcitonin-like peptide (CCLP) on osteoblast diff erentiation and mineralization in the culture system of MC3T3-E1 cells. Th e primary structures of the CCLP containing Cys-Asp-Gly-Val- Ser-Th r-Cys-Trp-Leu-His-Glu-Leu-Gly-Asn-Ser-Val-His-Ala-Th r-Ala-Gly-Gly-Lys-Gln-Asn-Val-Gly-Phe-Gly-Pro-NH2 was synthesized automatically using the solid phase method with fl uorenylmethoxycarbonyl (Fmoc) resin. Pre-osteoblast MC3T3-E1 cells were cultured with various concentrations of CCLP (7.5, 15, and 30 μM) during the osteoblast diff erentiation period. To examine osteoblast diff erentiation, alkaline phosphatase (ALP) activity was determined by reading the absorbance at 405 nm using a spectrophotometer, and mineralization was evaluated by staining with Alizalin red S. Moreover, the expression of diff erentiation markers such as ALP, osteocalcin (OSC), and osteopontin (OPN) were measured using RT-PCR and Western blot analysis. Th e results showed that CCLP did not exhibit any cytotoxic eff ect on MC3T3-E1 cells even at the highest concentration (30 μM) at 2 and 5 days. CCLP also enhanced MC3T3-E1 cells proliferation, diff erentiation, and mineralization demonstrated by the increased expression of several osteoblast phenotype markers such as ALP, and Alizarin red S staining. In addition, the CCLP induced mitogenactivated protein kinase (MAPK) pathway in MC3T3-E1 cells. Th ese results suggest that CCLP exerts positive eff ects on osteoblast diff erentiation and may represent a potential target for pharmaceutical development.