Table 1.

Enantioselectivities of l and d selective yeast-bound HRP variants toward 1 and 2 discovered in 3 rounds of directed evolution

HRP variant	Substrate	vl*	vd*	E(l/d)†
Wild-type	1	2.6 ± 0.6	1.6 ± 0.7	1.6 ± 0.5
Wild-type	2	21 ± 3	25 ± 4	0.8 ± 0.1
LIII‡	1	85 ± 6	1.8 ± 0.5	49 ± 1
LIII‡	2	4.2 ± 0.6	0.4 ± 0.1	10 ± 1
DIII§	1	0.8 ± 0.3	2.4 ± 0.8	0.3 ± 0.4
DIII§	2	2.0 ± 0.4	154 ± 4	0.013 ± 0.003

* v_l and v_d are the initial rates of oxidation of the l enantiomer and d enantiomer, respectively, reported in MFU/min. All experiments were conducted at least in triplicate with the mean and standard deviation values given in the table. See Methods for details.

^†Enantioselectivity, E(l/d), is defined as v_l/v_d. Note that E(l/d) × E(d/l) = 1.

^‡The l selective variant discovered in 3 rounds of directed evolution.

^§The d selective variant discovered in 3 rounds of directed evolution.