Fig. 4: Directed evolution of enzymes in IU pathway.
From: Yeast metabolism adaptation for efficient terpenoids synthesis via isopentenol utilization
a–c Homotrimeric structure of SmDAGK obtained by AlphaFold2, (b) shows the active site of SmDAGK, and (c) shows the cavity of SmDAGK. d Homodimer structure of AtIPK obtained from AlphaFold2. e Monomeric structure of AtIPK and its active site. f Schematic diagram of high-throughput screening method based on growth-coupled characteristics. g Conduct further verification of the promising single-point mutations obtained from SmDAGK and AtIPK, followed by combinatorial mutation. p = 0.002 between SmDAGKS47A, L124A + AtIPKS270P, A272R variants and control group compared with squalene titer. Statistical significance was evaluated by a two-tailed student’s t test (ns represents no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001). Data represent mean value with SD (three independent biological replicates). h, i Molecular docking results for wild-type enzyme and mutant. The substrate ATP and isoprenol and several key residues were shown. Source data are provided as a Source Data file.
