7B7H

The glucuronoyl esterase OtCE15A R268A variant from Opitutus terrae in complex with, and covalently linked to, D-glucuronate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	MAX IV BEAMLINE BioMAX
Synchrotron site	MAX IV
Beamline	BioMAX
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-02-23
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.008
Spacegroup name	P 1
Unit cell lengths	47.333, 47.298, 51.112
Unit cell angles	61.95, 67.92, 88.27

Refinement procedure

Resolution	43.150 - 1.800
R-factor	0.1851
Rwork	0.183
R-free	0.22240
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6gs0
RMSD bond length	0.007
RMSD bond angle	0.897
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]	43.150	1.861
High resolution limit [Å]	1.796	1.796
Rmerge	0.120	1.256
Number of reflections	32171	3093
<I/σ(I)>	6.84	0.95
Completeness [%]	96.6	92.99
Redundancy	3.6	3.6
CC(1/2)	0.995	0.461

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	298	Morpheus screen consisting of 0.1 M buffer system 1 (imidazole and MES), 30% precipitant mix 2 (ethylene glycol and PEG 8000) and amino acid additives (L-Glutamate, Alanine (racemic); Glycine; Lysine HCl (racemic); Serine (racemic)). Soaking was carried out in a saturated solution of Bnz-GlcA in 10% DMSO and 90% mother liquor for 30 seconds before being flash frozen.