Redesign of glycerol dehydratase from <i>L</i>. <i>reuteri</i>.

Abstract

<p>A) Overall fold of the glycerol dehydratase monomer and the proposed biological dimer; Stereodrawing of the B) 1,2-PD or C) glycerol bound form of the glycerol dehydratase. Homology modeling for the prediction of tertiary structure of glycerol dehydratase (PduCDE) resulted in a model based on the crystal structure of diol dehydratase-cyanocobalamin complex from <i>Klebsiella oxytoca</i> (1DIO), with sequence similarities and coverages above 42% and 95%, respectively. Quality model assessment of the homology model revealed a QMEAN6-score of 0.83 and a Z-score of -0.53. Ramachandran plot revealed that none of residues were present in the disallowed regions. Color codes for carbon atoms: yellow for adenosylcobalamin (AdoCbl), white for glycerol/1,2-PD, purple for calcium ion. The model figure was generated using Pymol. D) Sequence alignment of large subunit of glycerol dehydratase (PduC) from different bacteria. The numbering scheme follows the amino acid sequences of PduCs. Identical residues in all sequences are highlighted in black and conserved in grey. E) Relative activities of WT/mutant glycerol dehydratases towards different 1,2-diols/glycerol. The data is normalized by taking the activity of the wild-type enzyme towards glycerol as 100%.</p