High-level production of recombinant Arenicola marina globin chains in Escherichia coli: a new generation of blood substitute

Artif Cells Blood Substit Immobil Biotechnol. 2009;37(3):106-16. doi: 10.1080/10731190902908445.


This work reports for the first time the expression of a soluble B2 globin chain that is part of the extracellular hexagonal-bilayer haemoglobin from Arenicola marina. Two recombinant B2 globins were produced, one fused with gluthatione S-tranferase (B2-GST) and the other without a fusion tag (RecB2) and requiring a different purification procedure. We also describe a new method for the expression of globin that uses Studier's auto-induction medium together with the heme precursor delta-aminolevulinic acid. Media supplementation with the heme precursor delta-aminolevulinic acid in the culture increased heme synthesis by E. coli leading to the expression of the recombinant B2 globins in their active form. RecB2 and B2-GST were expressed with a yield of up to 105 mg/l of E. coli culture. Our approach is rapid and requires only one chromatographic purification step for B2-GST and three purification steps for RecB2. The overall results on RecB2 and B2-GST show that the recombinant globins exhibit similar properties to those of Arenicola marina native HBL-Hb with a great stability and a strong oxygen binding. The results and methodologies described in this paper are the beginning of a work aiming at reconstituting a recombinant HBL-Hb by genetic engineering in order to produce an innovative oxygen carrier for therapeutic applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Amino Acid Sequence
  • Aminolevulinic Acid / metabolism
  • Animals
  • Blood Substitutes / chemistry
  • Blood Substitutes / isolation & purification
  • Blood Substitutes / metabolism*
  • Escherichia coli / genetics*
  • Genetic Engineering / methods*
  • Globins / biosynthesis*
  • Globins / chemistry
  • Globins / isolation & purification
  • Globins / metabolism
  • Heme / metabolism
  • Hemoglobins / metabolism
  • Humans
  • Mass Spectrometry
  • Molecular Sequence Data
  • Oxidation-Reduction
  • Polychaeta*
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Solubility
  • Spectrophotometry, Ultraviolet


  • Blood Substitutes
  • Hemoglobins
  • Recombinant Fusion Proteins
  • Heme
  • Aminolevulinic Acid
  • Globins