8IBM

Sulfate bound form of PET-degrading cutinase Cut190 with thermostability-improving mutations of S226P/R228S/Q138A/D250C-E296C/Q123H/N202H and S176A inactivation

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PHOTON FACTORY BEAMLINE BL-17A
Synchrotron site	Photon Factory
Beamline	BL-17A
Temperature [K]	95
Detector technology	PIXEL
Collection date	2021-03-09
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9
Spacegroup name	P 3
Unit cell lengths	83.755, 83.755, 64.752
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	36.270 - 2.200
R-factor	0.2354
Rwork	0.234
R-free	0.26720
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.002
RMSD bond angle	0.478
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	37.000	2.330
High resolution limit [Å]	2.200	2.200
Number of reflections	25685	4113
<I/σ(I)>	10.4	1.4
Completeness [%]	99.6	98.5
Redundancy	5.5	5.5
CC(1/2)	0.997	0.743

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	293	0.1 M MES monohydrate pH 6.5, 0.2 M Ammonium sulfate, and 30% (w/v) PEG MME 5,000