Molecular genetic approaches to decrease the uncontrolled misincorporation of non-canonical branched chain amino acids into recombinant mini-proinsulin expressed in Escherichia coli.

coli strain showed that, mainly in the intracellular protein soluble protein fraction, norvaline and norleucine concentrations report a reduction compared with the reference strain under both tested cultivation conditions but effect of increasing L-arabinose concentrations did not show a clear trend on variation of ncBCAA concentrations.

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coli strains showing a preferred protein impurity profile during screening in the mini-reactor system were further verified in a 15 L reactor under cultivation conditions subjected to pyruvate pulsing and DO limitation, with the same recombinant mini-proinsulin production and a highly significant reduction of ncBCAA misincorporation into the recombinant protein in comparison with the non-engineered E. coli strain.

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