1OJJ

Anatomy of glycosynthesis: Structure and kinetics of the Humicola insolens Cel7BE197A and E197S glycosynthase mutants

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-1
Synchrotron site	ESRF
Beamline	ID14-1
Temperature [K]	100
Detector technology	CCD
Detector	MARRESEARCH
Spacegroup name	P 1 21 1
Unit cell lengths	66.364, 74.746, 85.820
Unit cell angles	90.00, 102.51, 90.00

Refinement procedure

Resolution	15.000 * - 1.400
R-factor	0.15
Rwork	0.149
R-free	0.17000 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1dym
RMSD bond length	0.011 *
RMSD bond angle	1.580 *
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	1.450
High resolution limit [Å]	1.400	1.400
Rmerge	0.033	0.300
Number of reflections	157771
<I/σ(I)>	35	3.5
Completeness [%]	99.0	96
Redundancy	3.6	3.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7		HANGING DROPS 20MM TRIS-HCL PH7-8.5, 280 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 (%)