3AI3

The crystal structure of L-Sorbose reductase from Gluconobacter frateurii complexed with NADPH and L-sorbose

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PHOTON FACTORY BEAMLINE AR-NE3A
Synchrotron site	Photon Factory
Beamline	AR-NE3A
Detector technology	CCD
Collection date	2009-10-30
Detector	ADSC QUANTUM 270
Wavelength(s)	1.000
Spacegroup name	P 21 21 21
Unit cell lengths	61.040, 124.410, 124.540
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.690 - 1.800
R-factor	0.21416
Rwork	0.212
R-free	0.24941
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3ai2
RMSD bond length	0.013
RMSD bond angle	1.476
Data reduction software	XDS
Data scaling software	XDS
Refinement software	REFMAC (5.5.0102)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	1.850
High resolution limit [Å]	1.800	1.800
Rmerge	0.083	0.465
Number of reflections	88419
<I/σ(I)>	21.3	5.6
Completeness [%]	99.8	100

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.5	293	Crystals of the His116Leu mutant of SR containing 10 mM NADPH and 100 mM L-sorbose were obtained under a reservoir solution condition of 30% (w/v) PEG400, 200 mM calcium chloride and 100 mM sodium acetate trihydrate, pH 4.5. The crystals of SR complexed with NADPH and L-sorbose were prepared by soaking the crystals in the reservoir solution supplemented with 2 M L-sorbose and 10 mM NADPH for 12 h at 293 K., VAPOR DIFFUSION, SITTING DROP, temperature 293.0K