1OJI

Anatomy of glycosynthesis: Structure and kinetics of the Humicola insolens Cel7B E197A and E197S glycosynthase mutants

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-4
Synchrotron site	ESRF
Beamline	ID14-4
Temperature [K]	100
Detector technology	CCD
Detector	ADSC CCD
Spacegroup name	P 65
Unit cell lengths	122.312, 122.312, 82.616
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	15.000 * - 2.150
R-factor	0.194
Rwork	0.192
R-free	0.22000 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1dym
RMSD bond length	0.018 *
RMSD bond angle	1.680 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	REFMAC (5.1.9999)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	15.000	2.230
High resolution limit [Å]	2.150	2.150
Rmerge	0.090	0.350
Number of reflections	38032
<I/σ(I)>	21	7
Completeness [%]	100.0	99.8 *
Redundancy	8.9	8.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7		HANGING DROPS 20MM TRIS-HCL PH7-8.5, 15-30% POLYETHYLENE GLYCOL 4000, pH 7.00

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	20 (mg/ml)
2	1	drop	Tris-HCl	20 (mM)	pH7-8.5
3	1	reservoir	PEG4000	15-30 (%)