The Protein Journal

, Volume 30, Issue 7, pp 464–470 | Cite as

Purification and Characterization of Albumin from Frog Skin of Duttaphrynus melanostictus

  • Ying-Xia Zhang
  • Cong-Wei Chen
  • Manchuriga Wang
  • Shuang-Shuang Wei
  • Huai Guan
  • Ting-Ting Chi
  • Xing-Zhu Qi
  • Wen-Ting Hu


Following determination of trypsin inhibitory activity, a serine protease inhibitor was purified and characterized from frog Duttaphrynus melanostictus serum. It was identified as serum albumin, with molecular weight of 67 kDa (DmA-serum). Different from bovine serum albumin, DmA-serum potently inhibited trypsin with similar K i values around 1.6 × 10−7 M. No inhibitory effect on thrombin, chymotrypsin, elastase and subtilisin was observed under the assay conditions. The N-terminal amino acid is EAEPHSRI. Subsequently, a protein with same N-terminal amino acid was purified from skin, termed as DmA-skin. However, DmA-skin is distinct from DmA-serum by binding of a haem b (0.5 mol/mol protein), and with low trypsin inhibitory activity. Frog albumin is distributed in frog skin and exhibited trypsin inhibitory activity, suggesting that it plays important roles in skin physiological functions, like water economy, metabolite exchange and osmoregulation, etc.


Albumin Skin Trypsin inhibitor Duttaphrynus melanostictus 



Serum albumin of Duttaphrynus melanostictus


Skin albumin of D. melanostictus










n-Benzoyl-l-tyrosine ethyl ester


3,3′-Diaminobenzidine tetrahydrochloride


Bovine serum albumin


Tris(hydroxy-methyl) aminomethane


Eathylene diamine tetraacetic acid


Sodium dodecyl sulfate polyacrylamide gel electrophoresis




Enzyme-linked immunosorbent assay


Phosphate buffer saline


Polyvinylidene fluoride


Horseradish peroxidase





This work was supported by the grants from the National Natural Science Foundation (30700128, 30960053), and Foundation of Hainan University (hd09xm67).


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ying-Xia Zhang
    • 1
    • 2
    • 5
  • Cong-Wei Chen
    • 1
    • 2
  • Manchuriga Wang
    • 3
  • Shuang-Shuang Wei
    • 3
  • Huai Guan
    • 4
  • Ting-Ting Chi
    • 1
    • 2
  • Xing-Zhu Qi
    • 1
    • 2
  • Wen-Ting Hu
    • 1
    • 2
  1. 1.Key Laboratory of Tropic Biological ResourcesMinister of EducationHaikouChina
  2. 2.Experimental Teaching Demonstration Center of Marine BiologyHainan UniversityHaikouChina
  3. 3.Hainan Key Laboratory of Tropical Animal Reproduction & Breeding and Epidemic Disease Research (Construction Period)Hainan UniversityHaikouChina
  4. 4.Haikou VTI Biological InstituteHaikouChina
  5. 5.College of Marine ScienceHainan UniversityHaikouPeople Republic of China

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