Fig. 5: Enhancing ALDH performance by protein engineering. | Nature Communications

Fig. 5: Enhancing ALDH performance by protein engineering.

From: RETRACTED ARTICLE: Systems engineering of Escherichia coli for high-level glutarate production from glucose

Fig. 5: Enhancing ALDH performance by protein engineering.

a Creation of the protein model introducing CYS residues (I90C, L91C, K92C, G210C, V211C, and I212C) visualized using Pymol. b. Glutarate production by different mutants under whole-cell conversion. Reactions were performed with recombinant E. coli (20 g/L whole cell catalyst) in 50 mL air-saturated PBS buffer (50 mM, pH 7.4) at 30 °C for 30 h (220 rpm). Glutarate titers were determined using HPLC. c Identification of residue sites in mutant Mu5 and its associated protein structure. d The distance between C1H, C5H, and NAD+ in both the WT and variant Mu5. e. DFT-computed Gibbs free energies (in kcal/mol) at the CPCM (water) level of theory and transition-state structures (Carbon: gray, hydrogen: white, Oxygen: red, Nitrogen: blue, angles are shown in o, and distances are shown in Å). The WT is shown in the black line, while mutant Mu5 is shown in the red line. n = 3 independent experiments. Data are presented as mean values ± SD. Source data are provided as a Source Data file.

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