9BJR

X-ray crystal structure of Y168F variant Thermothelomyces thermophilus polysaccharide monooxygenase 9E

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.1
Synchrotron site	ALS
Beamline	5.0.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-04-29
Detector	DECTRIS PILATUS 2M
Wavelength(s)	0.97741
Spacegroup name	C 2 2 21
Unit cell lengths	97.989, 119.970, 92.290
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	58.620 - 2.200
R-factor	0.1777
Rwork	0.175
R-free	0.21280
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	0.650
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]	58.620	2.270
High resolution limit [Å]	2.200	2.200
Rmerge	0.097	0.344
Rmeas	0.113	0.407
Rpim	0.057	0.211
Number of reflections	27208	2376
<I/σ(I)>	9.2	3.6
Completeness [%]	97.8	99.3
Redundancy	3.7	3.5
CC(1/2)	0.992	0.843

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, 35 % w/v polyethylene glycol 3,350). 1 ul 30 mg/mL 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 35% 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