5E4B

Hydroxynitrile lyase from the fern Davallia tyermanii in complex with (R)-mandelonitrile / benzaldehyde

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM14
Synchrotron site	ESRF
Beamline	BM14
Temperature [K]	100
Detector technology	CCD
Collection date	2014-06-22
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.9184
Spacegroup name	I 2 2 2
Unit cell lengths	73.362, 94.135, 116.143
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	35.800 - 1.500
R-factor	0.1573
Rwork	0.156
R-free	0.17900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5e46
RMSD bond length	0.007
RMSD bond angle	1.033
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	35.800	1.550
High resolution limit [Å]	1.500	1.500
Rmerge	0.066	0.611
Number of reflections	63070
<I/σ(I)>	13.74	1.86
Completeness [%]	97.5	80.88
Redundancy	4.1	3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	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).