Twenty sequences (plus associated wild type enzymes as controls) with models that most optimally recapitulated our desired active site geometry were chosen for additional experimental characterization by way of a pH profile. Proteins were produced and purified as described in the proceeding section, except that culture volumes were 20 mL, and assayed as described above.
Data are presented below as a table. In the table, the names of the substrates tested against are abbreviated. BEO = 2-benzyl ethylene oxide, PO = propene oxide, EO = ethylene oxide, and PR = propanal. No protein- and substrate-dependent activity on the target substrates was observed at any of the pHs tested, though protein yields and propanal activity broadly recapitulated earlier experiments.