1OJK

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	66.282, 74.830, 86.105
Unit cell angles	90.00, 102.57, 90.00

Refinement procedure

Resolution	30.000 * - 1.500
R-factor	0.143
Rwork	0.141
R-free	0.19000 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1dym
RMSD bond length	0.010 *
RMSD bond angle	1.560 *
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]	30.000	1.550
High resolution limit [Å]	1.500	1.500
Rmerge	0.043	0.320
Number of reflections	129121
<I/σ(I)>	24	3.8
Completeness [%]	99.3	97.4
Redundancy	3.5	3.6

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 (%)