5E46

Hydroxynitrile lyase from the fern Davallia tyermanii

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-05-09
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.979
Spacegroup name	I 2 2 2
Unit cell lengths	73.633, 94.020, 117.054
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	57.971 - 1.854
R-factor	0.1567
Rwork	0.155
R-free	0.19160
Structure solution method	SAD
RMSD bond length	0.007
RMSD bond angle	1.021
Data reduction software	XDS
Phasing software	PHENIX
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	58.000	1.920
High resolution limit [Å]	1.850	1.850
Rmerge	0.188	0.796
Number of reflections	34750
<I/σ(I)>	9.97	2.57
Completeness [%]	99.8	98.6
Redundancy	13.3	13

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	298	Native crystals of DtHNL1 were obtained by mixing 0.5ul 4 mg/mL protein sample (in 10 mM Tris-HCl pH 8.0) with 1 ul reservoir solution (0.9 M NaNO3; Na2HPO4; (NH4)2SO4 mix, 0.1 M Tris-Bicine Buffer pH 8.5 and 30% (w/v) polyethylene glycol monomethyl ether 550 & polyethylene glycol 20k; Morpheus condition C9). Additionally, native crystals were also grown by mixing 1 ul 4 mg/mL protein sample (in 10 mM Tris-HCl pH 8.0) with 0.5ul reservoir solution (0.1 M 2-(4-(2-hydroxyethyl)-1-piperazinyl) ethanesulfonic acid pH 7.5 and 10% (w/v) polyethylene glycol; JSCG condition B4). SeMet-DtHNL1 crystals were obtained in 0.2 M sodium thiocyanate, 20% (w/v) polyethylene glycol 3350. A 1:1 ratio of protein and screening solutions was used, using protein concentration of 3 mg/mL (in 10 mM Tris-HCl pH 8.0).