Correction to “Fortification of FeS Clusters Reshapes Anaerobic CO Dehydrogenase into an Air‐Viable Enzyme Through Multilayered Sealing of O₂ Tunnels”

Angew. Chem. Int. Ed. 2025, 64, e202508565, https://doi.org/10.1002/anie.202508565

In Table 1 of the main text and Table S5 of the Supporting Information, several kinetic parameters (k _cat, K _m , and k _cat/K _m ) contained numerical inconsistencies caused by rounding and formatting during data tabulation. All values have been recalculated using the original raw data, and the corrected tables are provided at the end of this document as corrected Table 1 and corrected Table S5.

Table 1.

Catalytic properties of ChCODH variants.

CODH variant	Substrate	Ni content a) (Ni mol/enzyme mol)	K m (mM)	k cat n b) (s−1)	k cat n/K m (s−1·mM−1)
ChCODH2	CO	0.42 ± 0.08	0.020 ± 0.002	3600 ± 700	181 000 ± 39 000
	EV		1.8 ± 0.1	2400 ± 460	1300 ± 260
A559H	CO	0.55 ± 0.05	0.061 ± 0.002	4200 ± 390	69 000 ± 6800
	EV		2.1 ± 0.1	4200 ± 400	2000 ± 200
A559W	CO	0.42 ± 0.02	0.033 ± 0.002	4300 ± 240	128 000 ± 12 000
	EV		2.0 ± 0.1	4800 ± 250	2400 ± 190
A559H/V610H	CO	0.70 ± 0.07	0.082 ± 0.008	4400 ± 490	54 000 ± 8100
	EV		1.7 ± 0.4	3100 ± 430	1800 ± 530
A559H/V610W	CO	0.50 ± 0.03	0.052 ± 0.012	3000 ± 350	57 000 ± 15 000
	EV		2.1 ± 0.3	3000 ± 250	1400 ± 230
A559W/V610H	CO	0.51 ± 0.01	0.091 ± 0.021	9200 ± 1100	100 000 ± 26 000
	EV		2.7 ± 0.5	7300 ± 610	2700 ± 580
A559W/V610W	CO	0.53 ± 0.01	0.058 ± 0.017	4900 ± 610	84 000 ± 27 000
	EV		1.5 ± 0.2	5500 ± 240	3800 ± 450

Kinetic data were assayed at 30 °C, pH 8. Parameters were estimated by nonlinear hyperbolic regression (SigmaPlot 10) of initial rates at seven EV concentrations (0.5–8 mM) and five CO concentrations (0.01–0.16 mM). All enzymatic specific activities (1 U mg⁻¹ = 1 µmol reduced viologen·min^‒1·mg^‒1) were determined in triplicate (see Methods). Most values are rounded to the nearest 100. The relative activities of WT, A559H, and A559W are from the same published data, reproduced with permission (ref. [8], Copyright 2022, Springer Nature).

^a) Ni contents refer to the molar ratio of nickel to enzyme based on total protein concentration. Values are the means ± standard deviation, n = 3.

^b) The normalized turnover number has been calculated by dividing the turnover by the Ni content.

Table S5.

Nickel content and kinetic parameters of ChCODH2 variants normalized to Ni content.

CODH variant	Substrate	Ni content (Ni mol/ Enzyme mol)	Specific activity a ) (U·mg−1)	Normalized Specific activity b ) (U·mg−1)	K m c ) (mM)	k cat d ) (s−1)	k cat n e ) (s−1)	k cat/K m (s−1·mM−1)	k cat n/K m (s−1·mM−1)
ChCODH2	CO	0.42 ± 0.08	1300 ± 108.5	3100 ± 258.9	0.020 ± 0.002	1500 ± 67	3600 ± 700	76 000 ± 7700	181 000 ± 39 000
	EV		900 ± 28.2	2150 ± 67.3	1.8 ± 0.1	1000 ± 16	2400 ± 460	540 ± 25	1300 ± 260
A559H	CO	0.55 ± 0.05	2100 ± 118.0	3840 ± 216	0.061 ± 0.002	2300 ± 38	4200 ± 390	38 000 ± 1500	69 000 ± 6800
	EV		1900 ± 63.5	3480 ± 116.3	2.1 ± 0.1	2300 ± 72	4200 ± 400	1100 ± 51	2000 ± 200
A559W	CO	0.42 ± 0.02	1800 ± 111.0	4300 ± 265.5	0.033 ± 0.002	1800 ± 50	4300 ± 240	54 000 ± 4200	128 000 ± 12 000
	EV		2000 ± 13.2	4780 ± 31.6	2.0 ± 0.1	2000 ± 39	4800 ± 250	1000 ± 62	2400 ± 190
A559H/V610H	CO	0.70 ± 0.07	2000 ± 23.1	2870 ± 33.1	0.082 ± 0.008	3100 ± 150	4400 ± 490	38 000 ± 4200	54 000 ± 8100
	EV		2000 ± 45.8	2870 ± 65.6	1.7 ± 0.4	2200 ± 210	3100 ± 430	1300 ± 350	1800 ± 530
A559H/V610W	CO	0.50 ± 0.03	1500 ± 3.5	2980 ± 6.9	0.052 ± 0.012	1500 ± 150	3000 ± 350	29 000 ± 7400	57 000 ± 15 000
	EV		1600 ± 17.3	3170 ± 34.3	2.1 ± 0.3	1500 ± 84	3000 ± 250	710 ± 110	1400 ± 230
A559W/V610H	CO	0.51 ± 0.01	2800 ± 81.9	5500 ± 161	0.091 ± 0.021	4700 ± 530	9200 ± 1100	51 000 ± 13 000	100 000 ± 26 000
	EV		2800 ± 53.3	5500 ± 104.8	2.7 ± 0.5	3700 ± 300	7300 ± 610	1400 ± 290	2700 ± 580
A559W/V610W	CO	0.53 ± 0.01	2100 ± 110.1	3990 ± 209.4	0.058 ± 0.017	2600 ± 320	4900 ± 610	45 000 ± 14 000	84 000 ± 27 000
	EV		2700 ± 28.9	5140 ± 55	1.5 ± 0.2	2900 ± 110	5500 ± 240	2000 ± 230	3800 ± 450

^a) Specific activity was measured as CO oxidation (or EV reduction) under standard assay conditions at 30 °C, pH 8.0.

^b) Normalized specific activity is the specific activity divided by Ni content.

^c) K _m was determined by nonlinear fitting of the Michaelis–Menten equation using five (CO) or seven (EV) substrate concentrations, calculated with SigmaPlot 10.

^d) k _cat was calculated from V _max values based on the enzyme concentration determined after Ni‐NTA purification.

^e) k _cat ⁿ is the Ni‐normalized turnover number, calculated by dividing k _cat by Ni content.

Most values are rounded to the nearest 100.

These updated values are also reflected in the following revised sentences on page 3 of the main text.

The original text,

“The normalized catalytic efficiency (k _cat ⁿ/K _m ) for CO was 100 000 s⁻¹·mM⁻¹ in A559W/V610H, comparable to 126 000 s⁻¹·mM⁻¹ in A559W. Similarly, A559H/V610H showed a catalytic efficiency of 55 000 s⁻¹·mM⁻¹ for CO”

Was updated as:

“…comparable to 128 000 s⁻¹·mM⁻¹ in A559W…”

“…showed a catalytic efficiency of 54 000 s⁻¹·mM⁻¹…”

These corrections do not affect the results or conclusions of the article.

The authors apologize for these errors.

Corrected Table 1 (main text)

Corrected Table S5 (Supporting Information)