3ZGJ

S221M V223F Y359A mutant of 4-Hydroxymandelate synthase from Streptomyces coelicolor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I02
Synchrotron site	Diamond
Beamline	I02
Temperature [K]	120
Detector technology	CCD
Collection date	2012-12-01
Detector	ADSC CCD
Spacegroup name	P 4
Unit cell lengths	122.870, 122.870, 54.860
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	54.950 - 1.950
R-factor	0.17711
Rwork	0.175
R-free	0.21504
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2r5v
RMSD bond length	0.021
RMSD bond angle	2.002
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	MOLREP
Refinement software	REFMAC (5.7.0032)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	61.440	2.010
High resolution limit [Å]	1.950	1.950
Rmerge	0.060	0.490
Number of reflections	57014
<I/σ(I)>	15.6	3.9
Completeness [%]	100.0	100
Redundancy	6.7	6.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		8.5		1 MICROLITRE 10 MG ML-1 HMS VARIANT, PREINCUBATED WITH 1.25 MM COCL2 AND 2.5 MM R-MANDELATE) PLUS 1 MICROLITRE MOTHER LIQUOR. THE BEST CRYSTALS WERE ROUTINELY OBTAINED IN CRYSTAL DROPS CONTAINING 0.05 M TRIS/CL PH 9.0, 0.1 M MGCL2, 7.5% (W/V) PEG 4K AND 2.5% (V/V) 2-METHYL-1,4-PENTANEDIOL.