Cloning, expression and antioxidant activity of a novel collagen from Pelodiscus sinensis
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Collagen is the main structural protein of various connective tissues in animals and naturally plays an important role within the body. It is increasingly used within certain areas, such as medicine, citology and cosmetology. The soft-shelled turtle (Pelodiscus sinensis) is a commercially important aquatic species rich in collagen. In this study, a novel collagen gene fragment of 756 bp, which encodes 252 deduced amino acid residues, including 25 conserved Gly-X-Y motifs, was cloned from a soft-shelled turtle. Recombinant soft-shelled turtle collagen (rSTC) was stably expressed in Escherichia coli Rosetta and purified by His GraviTrap affinity columns. The antioxidant activities of rSTC were measured using hydroxyl and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. The results showed that rSTC quenched the free radicals in a dose-dependent manner. The hydroxyl radical scavenging activity (HRSA) of rSTC was 98.9 % at a concentration of 3 mg/mL. At a concentration of 5 mg/mL, rSTC exhibited a DPPH radical scavenging activity of 32.7 %. At the tested concentrations, rSTC exhibited higher HRSA and lower DPPH radical scavenging activity.
KeywordsCollagen Cloning Expression Pelodiscus sinensis Antioxidant activity
This work was financially supported by the Spark Program of China (2014GA701059, 2014GA701018), the Open Foundation of Key Disciplines of Zhejiang Province (E02009124200), the Natural Science Foundation of Zhejiang Province (LY12C19004), the Science Foundation of the Department of Education of Zhejiang Province (Y201327815), and the Agricultural Program of Ningbo, Zhejiang, China (2011C10004), and was sponsored by the K. C. Wong Magna Fund of Ningbo University.
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