1GKL

S954A mutant of the feruloyl esterase module from clostridium thermocellum complexed with ferulic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SRS BEAMLINE PX14.2
Synchrotron site	SRS
Beamline	PX14.2
Temperature [K]	120
Detector technology	CCD
Collection date	2001-06-15
Detector	ADSC CCD
Spacegroup name	P 21 21 21
Unit cell lengths	65.101, 108.233, 112.964
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.920 - 1.400
R-factor	0.144
Rwork	0.143
R-free	0.16000
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.400 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	REFMAC (5.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	1.420
High resolution limit [Å]	1.400	1.400
Rmerge	0.048	0.136
Number of reflections	156561 *
<I/σ(I)>	33.5	6.6
Completeness [%]	99.6	94.5
Redundancy	5.2	2.6 *

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.5		HEPES PH 7.5 100 MM, NA ACETATE 1M, CD ACETATE 50 MM, GLYCEROL 5%

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	15 (mg/ml)
2	1	reservoir	sodium HEPES	0.1 (M)	pH7.5
3	1	reservoir	dithiothreitol	5 (mM)
4	1	reservoir		50 (mM)
5	1	reservoir	glycerol	5 (%(v/v))
6	1	reservoir	sodium acetate	1 (M)