African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12024

Full Length Research Paper

Expression and functional evaluation of Mytilus galloprovincialis foot protein type 5 (Mgfp-5), the recombinant mussel adhesive protein

Yawei Lv
  • Yawei Lv
  • Key Laboratory of Biotechnology, China College of Life Science, Northwest University, Shaanxi, 710069, Xiʼan, China.
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Yujing Zhang
  • Yujing Zhang
  • Key Laboratory of Biotechnology, China College of Life Science, Northwest University, Shaanxi, 710069, Xiʼan, China.
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Wenying Gao
  • Wenying Gao
  • Key Laboratory of Biotechnology, China College of Life Science, Northwest University, Shaanxi, 710069, Xiʼan, China.
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Yingjuan Wang
  • Yingjuan Wang
  • Key Laboratory of Biotechnology, China College of Life Science, Northwest University, Shaanxi, 710069, Xiʼan, China.
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  •  Received: 14 May 2017
  •  Accepted: 24 August 2017
  •  Published: 06 September 2017

Abstract

Mussel contains a variety of adhesion proteins, among which, Mytilus galloprovincialis foot protein type 5 (Mgfp-5) is one of the major proteins required for substrate adhesion. The labor-intensive nature and insufficiency of the extraction process have frequently resulted in very little purified recombinant Mgfp-5. These prompt technologies such as chemical synthesis and genetic engineering are employed to overcome these limitations. In this study, successful expression and purification of the recombinant Mgfp-5 using Escherichia coli BL21 (DE3) and affinity chromatography were reported. Production yield of 12.25% and purity of 96.92%, respectively were observed. The 3,4-dihydroxyphenylalanine (DOPA) content (9.60 pmol/g) and the adhesion (1 116 nN) in modified recombinant Mgfp-5 were 9.32 times and 1.6 times as great as those in the unmodified recombinant Mgfp-5, respectively. Recombinant Mgfp-5 at a concentration of 9.6 mg/L had little cytotoxicity on mouse L-929 fibroblast cells, which was toxic at first in cytotoxicity test, and a concentration of not more than 20 μg/mL would not lead to hemolysis of rabbit erythrocytes. In this case, recombinant Mgfp-5 is biosecure, providing the foundation for Mgfp-5 manufacturing as well as the development of clinical biological adhesive.

 

Key words: Mgfp-5, 3,4-dihydroxyphenylalanine (DOPA), adhesion, cytotoxicity, hemolysis.