9BJS

X-ray crystal structure of Y62W 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)	1.00003
Spacegroup name	C 2 2 21
Unit cell lengths	99.692, 122.395, 91.699
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	43.790 - 2.000
R-factor	0.1999
Rwork	0.199
R-free	0.23330
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.573
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]	43.790	2.050
High resolution limit [Å]	2.000	2.000
Rmerge	0.102	0.429
Rpim	0.042	0.175
Number of reflections	38124	2772
<I/σ(I)>	14.4	4.3
Completeness [%]	99.7	98.8
Redundancy	6.9	6.7
CC(1/2)	0.995	0.942

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, 25 % w/v polyethylene glycol 3,350). 1 ul 30 mg/mL Y62W 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 10 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