3 Biotech

, 8:181 | Cite as

Extraction and characterization of collagen hydrolysates from the skin of Rana chensinensis

  • Yuanyuan Zhao
  • Zhanyong Wang
  • Jing Zhang
  • Tingting Su
Original Article


The production of Hasma generates plentiful non-edible by-products in China and Central Asia. As one of main by-products, the skin of Rana chensinensis is discarded as waste without utilization. In this work, R. chensinensis skin collagen (RCSC) hydrolysates were extracted using pepsin under acidic conditions. The yield of RCSC is 15.1% (w/w). Amino acid analysis revealed that RCSC contained glycine (204.5/1000 residues) and imino acids (182/1000 residues). RCSC exhibited high solubility in acidic pH (1–4) and low NaCl concentrations (< 2%, w/v). Differential scanning calorimetry indicated that the denaturation temperature of RCSC was 33.5 °C. Scanning electron microscopy analysis confirmed their well-defined fibril morphologies. The results indicated that the skin of R. chensinensis is an alternative source of collagen hydrolysates, and RCSC can serve as a potential source applying in foodstuff and medical industry.


Rana chensinensis Skin Collagen hydrolysates Extraction Characterization 



The authors gratefully acknowledge the financial supports by Program for Liaoning Excellent Talents in University (Grant no. LJQ2014040) and Science and Technology Development Foundation of Fushun City (Grant no. 20142101).

Compliance with ethical standards

Conflict of interest

All the authors declare that they have no conflict of interest.


  1. Asghar A, Henrickson RL (1982) Chemical, biochemical, functional and nutritional characteristics of collagen in food systems. Adv Food Res 28:231–372CrossRefGoogle Scholar
  2. Doyle BB, Bendit EG, Blout ER (1975) Infrared spectroscopy of collagen and collagen-like polypeptides. Biopolymers 14:937–957CrossRefGoogle Scholar
  3. Duan R, Zhang J, Du X, Yao X, Konno K (2009) Properties of collagen from skin, scale and bone of carp (Cyprinus carpio). Food Chem 112:702–706CrossRefGoogle Scholar
  4. Foegeding EA, Lanier TC, Hultin HO (1996) Collagen. In: Fennema OR (ed) Food chemistry. Marcel Dekker, New York, pp 902–906Google Scholar
  5. Jongjareonrak A, Benjakul S, Visessanguan W, Nagai T, Tanaka M (2005a) Isolation and characterisation of acid and pepsin-solubilised collagens from the skin of brownstripe red snapper (Lutjanus vitta). Food Chem 93:475–484CrossRefGoogle Scholar
  6. Jongjareonrak A, Benjakul S, Visessanguan W, Tanaka M (2005b) Isolation and characterisation of collagen from bigeye snapper (Priacanthus marcracanthus) skin. J Sci Food Agr 85:1203–1210CrossRefGoogle Scholar
  7. Kaewdang O, Benjakul S, Kaewmanee T, Kishimura H (2014) Characteristics of collagens from the swim bladders of yellowfin tuna (Thunnus albacares). Food Chem 155:264–270CrossRefGoogle Scholar
  8. Kittiphattanabawon P, Benjakul S, Visessanguan W, Nagai T, Tanaka M (2005) Characterisation of acid-soluble collagen from skin and bone of bigeye snapper (Priacanthus tayenus). Food Chem 89:363–372CrossRefGoogle Scholar
  9. Kittiphattanabawon P, Benjakul S, Visessanguan W, Shahidi F (2010) Isolation and characterization of collagen from the cartilages of brown banded bamboo shark (Chiloscyllium punctatum) and blacktip shark (Carcharhinus limbatus). LWT Food Sci Technol 43:792–800CrossRefGoogle Scholar
  10. Komsa-Penkova R, Koyonava R, Kostov G, Tenchov B (1999) Discrete reduction of type I collagen thermal stability upon oxidation. Biophys Chem 83:185–195CrossRefGoogle Scholar
  11. Kumar B, Rani S (2017) Technical note on the isolation and characterization of collagen from fish waste material. J Food Sci Tech 54:276–278CrossRefGoogle Scholar
  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  13. Li H, Liu BL, Gao LZ, Chen HL (2004) Studies on bullfrog skin collagen. Food Chem 84:65–69CrossRefGoogle Scholar
  14. Liang Q, Wang L, Sun W, Wang Z, Xu J, Ma H (2014) Isolation and characterization of collagen from the cartilage of Amur sturgeon (Acipenser schrenckii). Process Biochem 49:318–323CrossRefGoogle Scholar
  15. Liu D, Zhou P, Li T, Regenstein JM (2014) Comparison of acid-soluble collagens from the skins and scales of four carp species. Food Hydrocoll 41:290–297CrossRefGoogle Scholar
  16. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Bio Chem 193:265–275Google Scholar
  17. Muyonga JH, Cole CGB, Duodu KG (2004) Characterisation of acid soluble collagen from skins of young and adult Nile perch (Lates niloticus). Food Chem 85:81–89CrossRefGoogle Scholar
  18. Nagai T, Suzuki N, Nagashima T (2008) Collagen from common minke whale (Balaenoptera acutorostrata). Food Chem 111:296–301CrossRefGoogle Scholar
  19. Nalinanon S, Benjakul S, Visessanguan W, Kishimura H (2007) Use of pepsin for collagen extraction from the skin of bigeye snapper (Priacanthus tayenus). Food Chem 104:593–601CrossRefGoogle Scholar
  20. Nalinanon S, Benjakul S, Kishimura H, Osako K (2011) Type I collagen from the skin of ornate threadfin bream (Nemipterus hexodon): characteristics and effect of pepsin hydrolysis. Food Chem 125:500–507CrossRefGoogle Scholar
  21. Pal GK, Suresh PV (2016) Sustainable valorisation of seafood by-products: recovery of collagen and development of collagen-based novel functional food ingredients. Innov Food Sci Emerg 37:201–215CrossRefGoogle Scholar
  22. Pal GK, Suresh PV (2017a) Comparative assessment of physico-chemical characteristics and fibril formation capacity of thermostable carp scales collagen. Mat Sci Eng C 70:32–40CrossRefGoogle Scholar
  23. Pal GK, Suresh PV (2017b) Physico-chemical characteristics and fibril-forming capacity of carp swim bladder collagens and exploration of their potential bioactive peptides by in silico approaches. Int J Biol Macromol 101:304–313CrossRefGoogle Scholar
  24. Pal GK, Nidheesh T, Suresh PV (2015) Comparative study on characteristics and in vitro fibril formation ability of acid and pepsin soluble collagen from the skin of catla (Catla catla) and rohu (Labeo rohita). Food Res Int 76:804–812CrossRefGoogle Scholar
  25. Pal GK, Nidheesh T, Govindaraju K, Jyoti Suresh PV (2017) Enzymatic extraction and characterisation of a thermostable collagen from swim bladder of rohu (Labeo rohita). J Sci Food Agric 97:1451–1458CrossRefGoogle Scholar
  26. Payne KJ, Veis A (1988) Fourier transform IR spectroscopy of collagen and gelatin solutions: deconvolution of the amide I band for conformational studies. Biopolymers 27:1749–1760CrossRefGoogle Scholar
  27. Purna Sai K, Babu M (2001) Studies on Rana tigerina skin collagen. Comp Biochem Physiol B Biochem Mol Biol 128:81–90CrossRefGoogle Scholar
  28. Rochdi A, Foucat L, Renou JP (2000) NMR and DSC studies during thermal denaturation of collagen. Food Chem 69:295–299CrossRefGoogle Scholar
  29. Schuetz T, Richmond N, Harmon ME, Schuetz J, Castaneda L, Slowinska K (2013) The microstructure of collagen type I gel cross-linked with gold nanoparticles. Colloids Surf B Biointerfaces 101:118–125CrossRefGoogle Scholar
  30. Singh P, Benjakul S, Maqsood S, Kishimura H (2011) Isolation and characterisation of collagen extracted from the skin of striped catfish (Pangasianodon hypophthalmus). Food Chem 124:97–105CrossRefGoogle Scholar
  31. Surewicz WK, Mantsch HH (1988) New insight into protein secondary structure from resolution enhanced infrared spectra. Biochim Biophys Acta 952:115–130CrossRefGoogle Scholar
  32. Uriarte-Montoya MH, Arias-Moscoso JL, Plascencia-Jatomea M, Santacruz-Ortega H, Rouzaud-Sández O, Cardenas-Lopez JL, Marquez-Rios E, Ezquerra-Brauer JM (2010) Jumbo squid (Dosidicus gigas) mantle collagen: extraction, characterization, and potential application in the preparation of chitosan–collagen biofilms. Bioresour Technol 101:4212–4219CrossRefGoogle Scholar
  33. Veeruraj A, Arumugam M, Ajithkumar T, Balasubramanian T (2012) Isolation and characterization of drug delivering potential of type-I collagen from eel fish Evenchelys macrura. J Mater Sci Med 23:1729–1738CrossRefGoogle Scholar
  34. Yin YG, Han YZ, Han Y (2006) Pulsed electric field extraction of polysaccharide from Rana temporaria chensinensis David. Int J Pharm 312:33–36CrossRefGoogle Scholar
  35. Zeng SK, Yin JJ, Yang SQ, Zhang CH, Yang P, Wu WL (2012) Structure and characteristics of acid and pepsin-solubilized collagens from the skin of cobia (Rachycentro canadum). Food Chem 135:1975–1984CrossRefGoogle Scholar
  36. Zhang J, Duan R (2017) Characterisation of acid-soluble and pepsin-solubilised collagen from frog (Rana nigromaculata) skin. Int J Biol Macromol 101:638–642CrossRefGoogle Scholar
  37. Zhang Y, Liu WT, Li GY, Shi B, Miao YQ, Wu XH (2007) Isolation and partial characterization of pepsin-soluble collagen from the skin of grass carp (Ctenopharyngodon idella). Food Chem 103:906–912CrossRefGoogle Scholar
  38. Zhao J, Sun Y, Li Z, Su Q (2011) Molecular cloning of novel antimicrobial peptide genes from the skin of the Chinese brown frog, Rana chensinensis. Zoolog Sci 28:112–117CrossRefGoogle Scholar
  39. Zhu R, Shi S (2013) Collagen extracted from fresh skin of Rana chensinensis by acetic acid and pepsin successively. J Yanbian Univ (Nat Sci Ed) 39:202–205Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Chemistry, Chemical Engineering and Environmental EngineeringLiaoning Shihua UniversityFushunChina

Personalised recommendations