High-level Production of Recombinant Arenicola Marina Globin Chains in Escherichia Coli: A New Generation of Blood Substitute - Adaptation et Biologie des Invertébrés en Conditions Extrêmes Accéder directement au contenu
Article Dans Une Revue Artificial Cells, Blood Substitutes and Biotechnology Année : 2009

High-level Production of Recombinant Arenicola Marina Globin Chains in Escherichia Coli: A New Generation of Blood Substitute

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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 -aminolevulinic acid. Media supplementation with the heme precursor -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.
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hal-01250943 , version 1 (28-04-2020)

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Thomas Harnois, Morgane Rousselot, Hélène Rogniaux, Franck Zal. High-level Production of Recombinant Arenicola Marina Globin Chains in Escherichia Coli: A New Generation of Blood Substitute. Artificial Cells, Blood Substitutes and Biotechnology, 2009, 37 (3), pp.106-116. ⟨10.1080/10731190902908445⟩. ⟨hal-01250943⟩
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