9BJT

X-ray crystal structure of Y62F Thermothelomyces thermophilus polysaccharide monooxygenase 9E

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-12-15
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.00004
Spacegroup name	C 2 2 21
Unit cell lengths	107.894, 114.606, 91.417
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.510 - 3.000
R-factor	0.2299
Rwork	0.226
R-free	0.26260
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.093
Data reduction software	iMOSFLM
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	39.510	3.180
High resolution limit [Å]	3.000	3.000
Rmerge	0.341	0.572
Rpim	0.205	0.344
Number of reflections	11339	1779
<I/σ(I)>	3.1	1.9
Completeness [%]	90.4	96.3
Redundancy	3.1	3.3
CC(1/2)	0.839	0.680

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	293	The crystal was grown by hanging drop in conditions optimized from Hampton Index HT G8 (0.2 M Ammonium acetate, 0.1 M HEPES pH 7.5, 28 % w/v polyethylene glycol 3,350). 1 ul 28 mg/mL Y62F Mt PMO9E in 50 mM MOPS, 50 mM MES was mixed with 1 uL with 0.2 M Ammonium acetate, 0.1 M HEPES pH 7.5, and 26% PEG 3,350. The crystal was then cryoprotected for 6 seconds in well solution mixed with cryoprotectants to a final concentration of 8% w/v sucrose, 2% w/v glucose, 8% v/v glycerol, and 8% ethylene glycol