6YTT

CO-dehydrogenase/Acetyl-CoA synthase (CODH/ACS) from Clostridium autoethanogenum at 3.0-A resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P13 (MX1)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-05-26
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.74281
Spacegroup name	P 42 21 2
Unit cell lengths	298.943, 298.943, 128.895
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.146 - 3.010
R-factor	0.1932
Rwork	0.192
R-free	0.22010
Structure solution method	SAD
Data reduction software	XDS
Data scaling software	STARANISO
Phasing software	AutoSol
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.150	3.315
High resolution limit [Å]	3.007	3.007
Rmerge	0.162	1.976
Rmeas	0.168	2.035
Rpim	0.043	0.485
Number of reflections	81409	4070
<I/σ(I)>	14.2	1.7
Completeness [%]	96.5	78.2
Redundancy	15	17.5
CC(1/2)	0.998	0.646

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	291.15	All crystallization were performed in anoxic chamber (N2/H2, 95:5 %) with anaerobic solutions. Crystals were obtained by initial screening at 291.15 K using the sitting drop method on a 96-well MRC 2 Crystallization Plates in polystyrene (SWISSCI). The crystallization reservoir contained 90 uL of mother liquor, crystallization drop contained a mixture of 0.6 uL protein and 0.6 uL precipitant. The best diffracting crystal of CaCODH/ACS from heterotrophic growth on fructose was obtained by initial screening using the Wizard 3 & 4 screen from Jena Bioscience. The crystallization reservoir contained 100 mM BIS-TRIS propane, pH 6.5; 200 mM Sodium fluoride and 20% (w/v) Polyethylene glycol 3,350. The initial protein concentration was 16.4 mg/mL.