9PV1

Biotin halogenase BtnX, anaerobic structure with Fe(II), biotin, alpha-ketoglutarate, chloride

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.3.1
Synchrotron site	ALS
Beamline	8.3.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2025-05-07
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	1.11
Spacegroup name	P 21 21 21
Unit cell lengths	50.868, 82.775, 173.355
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	86.680 - 1.200
R-factor	0.148
Rwork	0.147
R-free	0.16960
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.073
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	86.680	1.243
High resolution limit [Å]	1.200	1.200
Rmerge	0.070	0.993
Rpim	0.021	0.414
Number of reflections	222979	18318
<I/σ(I)>	16.89	1.66
Completeness [%]	97.6
Redundancy	11.7
CC(1/2)	0.999	0.722

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	298	1:1 v/v mixing of crystallographic solution (26% w/v PEG4000, 0.1 M TRIS, pH 8.0, 0.2 M lithium sulfate) with protein solution (10 mg/mL purified BtnX, 1.0 mM alpha-ketoglutarate, 1.75 mM biotin, 50 mM NaCl). After crystallization 0.5 M of (NH4)2Fe(SO4)2 was added, followed by 30% v/v ethylene glycol