. Author manuscript; available in PMC: 2023 Jan 14.

Published in final edited form as: J Am Chem Soc. 2022 Dec 21;145(1):537–550. doi: 10.1021/jacs.2c10775

Table 2.

Activity and selectivity of representative engineered CYP119 variants for the intermolecular C─H functionalization of N-phenylpyrrolidine with EDA.^[a]


Entry	Enzyme Variant	Yield^[b]	TON^[c]	e.r. (6a:7a)
1	CYP119 (WT)	3%	76	n.d.
2	CYP119(C317S)	5%	110	n.d.
3	CYP119(T213A	6%	135	n.d.
4	CYP119(T213A, C317S	18%	812	71:29
5	CYP119(T213A, V254A, C317S) (CHI-g1)	99%	4,400	79:21
6	CYP119(F153G, T213A, V254A, C317S) (CHI-EDA)	89% (64%)^[d]	8,920	87:13
7^[e]	CYP119(F153G, T213A, V254A, C317S)	99%	500	87:13
8^[f]	CYP119(F153G, T213A, V254A, C317S)	87%	8,710	87:13
9^{[f, g]}	CYP119(F153G, T213A, V254A, C317S)	51%	5,090	87:13
10	CYP119(A209W, T213G, V254A, C317S	71%	6,750	19:81

^[a]

Standard reaction conditions: CYP119 expressing C41(DE3) E. coli cells, OD₆₀₀ = 40, 10 mM 1a, 20 mM EDA, in KPi buffer (50 mM, pH 7), room temperature, 16 hours, in anaerobic chamber.

^[b]

Assay yields as determined by GC using calibration curves generated with isolated product.

^[c]

TON as calculated based on the protein concentration of the cell lysate.

^[d]

Isolated yield.

^[e]

Using 20 μM purified protein and 10 mM Na₂S₂O₄.

^[f]

Reaction time: 2 hours.

^[g]

Under aerobic conditions. N.d. = not determined.