Representative variants underlining the directed evolution of hPGK- serine 219.

Noted are sequence changes, growth rates and kcat/KM values of representative variants along the laboratory trajectory. Variants annotation: G stands for the round number from which a variant was isolated, and v denotes the variant's sequential number within this round. The left column denotes the sequence composition and rate parameters of wild-type hPGK. In bold are mutations that appeared at ≥90% of the variants by the last round (near complete fixation). Mutations that were not fixed appear in standard letters and their percentage of occurrence is denoted in brackets (no percentage is indicated for mutations that appeared only once in >100 variants).aNote that kcat/KM for wild type hPGK we measured was ∼11-fold lower than previously reported [18]. The origins of this difference are unknown, yet the rates for various mutants were measured by us under the very same conditions as wild-type, thus providing consistency within the reported set. The ± mark represents the S.D within 3 independent measurements.