Advertisement

Cloning, expression and antioxidant activity of a novel collagen from Pelodiscus sinensis

  • Ran Xu
  • Dengfeng Li
  • Jiao Peng
  • Jing Fang
  • Liping Zhang
  • Lianguo Liu
Original Paper

Abstract

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.

Keywords

Collagen Cloning Expression Pelodiscus sinensis Antioxidant activity 

Notes

Acknowledgments

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.

References

  1. Adachi T, Tomita M, Shimizu K, Ogawa S, Yoshizato K (2006) Generation of hybrid transgenic silkworms that express Bombyx mori prolyl-hydroxylase α-subunits and human collagens in posterior silk glands: production of cocoons that contained collagens with hydroxylated proline residues. J Biotechnol 126:205–219CrossRefGoogle Scholar
  2. Bi L (2006) Study on physicochemical characters and biological activities of collagen from the body of sea cucumber (Stichopus japonicus). Dissertation, Ocean University of ChinaGoogle Scholar
  3. Buechter DD, Paolella DN, Leslie BS, Brown MS, Mehos KA, Gruskin EA (2003) Co-translational incorporation of trans-4-hydroxyproline into recombinant proteins in bacteria. J Biol Chem 278:645–650CrossRefGoogle Scholar
  4. Cacciuttolo MA, Trinh L, Lumpkin JA, Rao G (1993) Hyperoxia induces DNA damage in mammalian cells. Free Rad Biol Med 14:267–276CrossRefGoogle Scholar
  5. Chen HH, Yu P, Li JR (2010) Preparation and free radical scavenging ability of collagen polypeptide isolated from Acaudina molpadioides Semper. J Chinese Inst Food Sci Technol 10:19–25Google Scholar
  6. Duan R, Zhang JJ, Du XQ, Yao XC, Konno K (2009) Properties of collagen from skin, scale and bone of carp (Cyprinus carpio). Food Chem 112:702–706CrossRefGoogle Scholar
  7. Hiruma H, Toida H, Hanawa T, Sakuragi H, Suzuki Y (2011) Biocompatibility control of recombinant collagen by ion beam modification. Surf Coat Tech 206:911–915CrossRefGoogle Scholar
  8. Hu WT, Sun M, Wang YJ (2006) Isolation and characterization of antioxidant peptide from scallop Chlamys Farreri. Oceanologia ET Limnlolgia Sinica 37:14–19Google Scholar
  9. Je JY, Qian ZJ, Byun HG, Kim SK (2007) Purification and characterization of an antioxidant peptide obtained from tuna backbone protein by enzymatic hydrolysis. Process Biochem 42:840–846CrossRefGoogle Scholar
  10. Jeong TH, Son YJ, Ryu HB, Koo BK, Jeong SM, Hoang P, Do BH, Song JA, Chong SH, Robinson RC, Choe H (2014) Soluble expression and partial purification of recombinant human erythropoietin from E. coli. Protein Expres Purif 95:211–218CrossRefGoogle Scholar
  11. Li Z, Wang B, Zhang Q, Qu Y, Xu H, Li G (2012) Preparation and antioxidant property of extract and semipurified fractions of Caulerpa racemosa. J Appl Phycol 24:1527–1536CrossRefGoogle Scholar
  12. Li ZR, Wang B, Chi CF, Gong YD, Luo HY, Ding GF (2013) Influence of average molecular weight on antioxidant and functional properties of cartilage collagen hydrolysates from Sphyrna lewini, Dasyatis akjei and Raja porosa. Food Res Int 51:283–293CrossRefGoogle Scholar
  13. Liang QF, Wang L, He YQ, Wang ZB, Xu JM, Ma HL (2014a) Hydrolysis kinetics and antioxidant activity of collagen under simulated gastrointestinal digestion. J Funct Foods 11:493–499CrossRefGoogle Scholar
  14. Liang QF, Wang L, Sun WH, Wang ZB, Xu JM, Ma HL (2014b) Isolation and characterization of collagen from the cartilage of Amur sturgeon (Acipenser schrenckii). Process Biochem 49:318–323CrossRefGoogle Scholar
  15. Lou JW, Zhu L, Wu MB, Yang LR, Lin JP, Cen PL (2014) High-level soluble expression of the hemA gene from Rhodobacter capsulatus and comparative study of its enzymatic properties. J Zhejiang Univ Sci B 15:491–499CrossRefGoogle Scholar
  16. Lu JF, Wan Q, Yin ZM, Lin L, Weng SB, Ye YW, Jiang ST (2010) Extraction and characterization of collagen from calipash of Chinese soft-shelled turtle (Pelodiscus sinensis). J Fish China 34:981–988CrossRefGoogle Scholar
  17. Nagai N, Kobayashi H, Katayama S, Munekata M (2009) Preparation and characterization of collagen from soft-shelled turtle (Pelodiscus Sinensis) skin for biomaterial applications. J Biomat Sci-Polym E 20:567–576CrossRefGoogle Scholar
  18. Ren JY, Zhao MM, Shi J, Wang JS, Jiang YM, Cui C, Kakuda Y, Xue SJ (2008) Purification and identification of antioxidant peptides from grass carp muscle hydrolysates by consecutive chromatography and electrospray ionization-mass spectrometry. Food Chem 108:727–736CrossRefGoogle Scholar
  19. Ruan Z, Liu G, Wang B, Zhou Y, Lu J, Wang Q, Zhao J, Zhang L (2014) First report of a peroxiredoxin homologue in jellyfish: molecular cloning, expression and functional characterization of CcPrx4 from Cyanea capillata. Mar Drugs 12:214–231CrossRefGoogle Scholar
  20. Song W, Chen W, Yang YN, Li CY, Qian GY (2014) Extraction optimization and characterization of collagen from the lung of soft-shelled turtle Pelodiscus sinensis. Int J Nutr Food Sci 3:270–278CrossRefGoogle Scholar
  21. Suetsuna K, Ukeda H, Ochi H (2000) Isolation and characterization of free radical scavenging activities peptides derived from casein. J Nutr Biochem 11:128–131CrossRefGoogle Scholar
  22. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 10:2731–2739. doi: 10.1093/molbev/msr121 CrossRefGoogle Scholar
  23. Tsai IJ, Zarowiecki M, Holroyd N, Garciarrubio A, Sanchez-Flores A, Brooks KL, Tracey A, Bobes RJ, Fragoso G, Sciutto E, Aslett M, Beasley H, Bennett HM, Cai J, Camicia F, Clark R, Cucher M, De Silva N, Day TA, Deplazes P, Estrada K, Fernández C, Holland PW, Hou J, Hu S, Huckvale T, Hung SS, Kamenetzky L, Keane JA, Kiss F, Koziol U, Lambert O, Liu K, Luo X, Luo Y, Macchiaroli N, Nichol S, Paps J, Parkinson J, Pouchkina-Stantcheva N, Riddiford N, Rosenzvit M, Salinas G, Wasmuth JD, Zamanian M, Zheng Y, Taenia solium Genome Consortium, Cai X, Soberón X, Olson PD, Laclette JP, Brehm K, Berriman M (2013) The genomes of four tapeworm species reveal adaptations to parasitism. Nature 496:57–63CrossRefGoogle Scholar
  24. Veeruraj A, Arumugam M, Balasubramanian T (2013) Isolation and characterization of thermostable collagen from the marine eel-fish (Evenchelys macrura). Process Biochem 48:1592–1602CrossRefGoogle Scholar
  25. Wang B, Wang YM, Chi CF, Luo HY, Deng SG, Ma JY (2013) Isolation and characterization of collagen and antioxidant collagen peptides from scales of Croceine Croaker (Pseudosciaena crocea). Mar Drugs 11:4641–4661CrossRefGoogle Scholar
  26. Wang J, Qi ZH, Yang ZC (2014) Evaluation of the protein requirement of juvenile Chinese soft-shelled turtle (Pelodiscus sinensis, Wiegmann) fed with practical diets. Aquaculture 433:252–255CrossRefGoogle Scholar
  27. Wu J, Lei JY, Zhang LF, Hua H, Jin J (2008) Improving production and bioactivity of recombinant human disintegrin domain of ADAM15 (rhADAM15) in Escherichia Coli. Acta Microbiologica Sinica 48:1067–1074Google Scholar
  28. Young ND, Jex AR, Li B, Liu S, Yang L, Xiong Z, Li Y, Cantacessi C, Hall RS, Xu X, Chen F, Wu X, Zerlotini A, Oliveira G, Hofmann A, Zhang G, Fang X, Kang Y, Campbell BE, Loukas A, Ranganathan S, Rollinson D, Rinaldi G, Brindley PJ, Yang H, Wang J, Wang J, Gasser RB (2012) Whole-genome sequence of Schistosoma haematobium. Nat Genet 44:221–225CrossRefGoogle Scholar
  29. Zeng MY, Guo Y, Liu ZY, Dong SY, Zhao YH, Cui WX (2008) Preparation of radical scavenging peptides from Oreochromis niloticus skin gelatin and its antioxidant activity. J Fish China 32:117–123Google Scholar
  30. Zheng H, Zhang W, Zhang L, Zhang Z, Li J, Lu G, Zhu Y, Wang Y, Huang Y, Liu J, Kang H, Chen J, Wang L, Chen A, Yu S, Gao Z, Jin L, Gu W, Wang Z, Zhao L, Shi B, Wen H, Lin R, Jones MK, Brejova B, Vinar T, Zhao G, McManus DP, Chen Z, Zhou Y, Wang S (2013) The genome of the hydatid tapeworm Echinococcus granulosus. Nat Genet 45:1168–1175CrossRefGoogle Scholar
  31. Zhu XF (2008) The experimental guide for gene engineering. Higher Education Press, BeijingGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Ran Xu
    • 1
  • Dengfeng Li
    • 1
  • Jiao Peng
    • 1
  • Jing Fang
    • 1
  • Liping Zhang
    • 1
  • Lianguo Liu
    • 1
  1. 1.Department of Marine SciencesNingbo UniversityNingboChina

Personalised recommendations