Fig. 3: Rational design of inefficient enzymes PdxA and PdxJ.

a The PN titer and cell growth (OD₆₀₀) of WT enzyme (Con.) and pdxA mutants. Data are presented as mean values ± SD from three independent biological replicates (n = 3), the circles or diamonds represent individual data points. b Comparison of the binding conformations of 4HTP in WT and H136N based on molecular dynamics (MD) analysis. The dotted green line indicates hydrogen bonding interactions. c Residues (yellow sticks) within a 6 Å radius from the imine intermediate P30 (cyan sticks). d The PN titer distribution of four rounds of PdxJ mutants including a total of 212 mutants: 26 single mutants (round 1), 62 double mutants (round 2), 60 triple mutants (round 3), and 64 quadruple mutants (round 4). All the detailed data are listed in Supplementary Data 3. e Distribution of the mutation sites (E104T/G194C/I218L) in LL239. f and g, h and i, j and k Differences before and after introducing the E104T, G194C, and I218L mutations, respectively. Substrates and residues are represented as pink in the WT and green in mutant structures. The dotted green line indicates hydrogen bonding interaction and the solid blue line indicates hydrophobic interaction. Source data are provided as a Source Data file.