Comparison of recombinant α-hemoglobin from Crocodylus siamensis expressed in different cloning vectors and their biological properties

Protein Expr Purif. 2016 Feb:118:55-63. doi: 10.1016/j.pep.2015.09.028. Epub 2015 Oct 9.

Abstract

Hemoglobin (Hb) is an important component in red blood cells of the vertebrate. It is a major respiratory protein with oxygen or carbon dioxide transport function. Hb has been reported to contain bioactive peptides which have antibacterial and antioxidant activities. In this study, the alpha-chain hemoglobin(αHb) gene of Crocodylus siamensis was cloned into the three different expression vectors and expressed in Escherichia coli BL21 (DE3). The recombinant αHb proteins from all constructs could be expressed and purified. The result from UV-visible absorption spectra showed a similar pattern of all recombinant proteins to the oxy-hemoglobin form of intact Hb. The different recombinant αHb could exhibit antioxidant activities. All recombinant proteins could inhibit the growth of Bacillus spp. Especially, most of the recombinant proteins could inhibit the growth of Bacillus amyloliquefaciens TISTR 1045 better than intact one. The result obtained from this study can provide us further information about the possibility using of αHb as a supplementary food.

Keywords: Alpha hemoglobin; Crocodylus siamensis; Protein expression; Protein purification; Recombinant protein.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alligators and Crocodiles / genetics*
  • Amino Acid Motifs
  • Animals
  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / metabolism
  • Anti-Bacterial Agents / pharmacology
  • Bacillus / drug effects
  • Bacillus / growth & development
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression*
  • Genetic Vectors / genetics*
  • Genetic Vectors / metabolism
  • Hemoglobins / chemistry
  • Hemoglobins / genetics*
  • Hemoglobins / metabolism
  • Hemoglobins / pharmacology*
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics*
  • Peptide Fragments / metabolism
  • Peptide Fragments / pharmacology*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Recombinant Proteins / pharmacology

Substances

  • Anti-Bacterial Agents
  • Hemoglobins
  • Peptide Fragments
  • Recombinant Proteins
  • alpha(A) globin