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. 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 1.

Intermolecular C─H functionalization of N-phenylpyrrolidine (2a) with diazoacetone using hemoproteins and variants thereof.[a]

graphic file with name nihms-1861408-t0009.jpg
Entry Catalyst Yield[b] TON[c] e.r.[d]
(3a:4a)
1 Hemin 0% 0 -
2 Mb (WT) 0% 0 -
3 Mb(H64G, V68A) 0% 0 -
4 P411-CHF 0% 0 -
5 CYP119 (WT) 0% 0 -
6 CYP119 (T213A) 0.4% 35 n.d.
7 CYP119(T213A, C317S) 2% 170 69:31
8 CYP119 (T213A, V254A, C317S) CHI-g1 14% 200 94:6
9 CYP119 (A209G, T213A, V254A, C317S) CHI-g2 23% 3,100 99.5:0.5
10 CYP119 (F153G, A209G, T213A, V254A, C317S) CHI-DA 99% 12,900 99.5:0.5
11[e] CYP119 (F153G, A209G, T213A, V254A, C317S) CHI-DA 99% 500 99.5:0.5
12[f] CYP119 (F153G, A209G, T213A, V254A, C317S) CHI-DA 53% 20,350 99.5:0.5
[a]

Standard reaction conditions: protein expressing C41(DE3) E. coli cells, OD600 = 40, 10 mM 2a, 20 mM diazoacetone (1a), in KPi buffer (50 mM, pH 7), room temperature, 16 hours, in anaerobic chamber.

[b]

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

[c]

TON as calculated based on the protein concentration measured from cell lysate.

[d]

Enantiomeric ratio (e.r.) for 3a:4a as determined by chiral SFC.

[e]

Using 20 μM purified protein and 10 mM Na2S2O4.

[f]

OD600 = 10. N.d. = not determined.