Journal article

Optimised expression and purification of recombinant human indoleamine 2,3-dioxygenase

CJD Austin, J Mizdrak, A Matin, N Sirijovski, P Kosim-Satyaputra, RD Willows, TH Roberts, RJW Truscott, G Polekhina, MW Parker, JF Jamie

PROTEIN EXPRESSION AND PURIFICATION | ACADEMIC PRESS INC ELSEVIER SCIENCE | Published : 2004

Abstract

The hemoprotein indoleamine 2,3-dioxygenase (IDO) is the first and rate-limiting enzyme in mammalian tryptophan metabolism. It has received considerable attention in recent years, particularly due to its role in the pathogenesis of many diseases. Here, we report attempts to improve soluble expression and purification of hexahistidyl-tagged recombinant human IDO from Escherichia coli (EC538, pREP4, and pQE9-IDO). Significant formation of inclusion bodies was noted at the growth temperature of 37 degrees C, with reduced formation at 30 degrees C. The addition of the natural biosynthetic precursor of protoporphrin IX, delta-aminolevulinic acid (ALA), coupled with optimisation of IPTG induction ..

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