6O1A

Alpha-L-fucosidase AlfC from Lactobacillus casei in complex with alpha-L-fucose product

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-11-01
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.033202
Spacegroup name	C 2 2 21
Unit cell lengths	89.983, 138.236, 264.440
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.840 - 2.600
R-factor	0.2087
Rwork	0.208
R-free	0.22620
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6o18
RMSD bond length	0.005
RMSD bond angle	1.269
Data reduction software	XDS
Data scaling software	Aimless (0.5.32)
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0238)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	29.840	29.820	2.680
High resolution limit [Å]	2.600	10.720	2.600
Rmerge	0.056	0.035	1.292
Rmeas	0.063	0.039	1.437
Rpim	0.028	0.017	0.619
Number of reflections	50905	759	4390
<I/σ(I)>	13.4
Completeness [%]	99.7	93.8	100
Redundancy	4.8	4.5	5.1
CC(1/2)	0.998	0.998	0.484

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	298	250 nL 20 mg/mL wild-type AlfC + 250 nL mother liquor (18% w/v PEG3350, 0.1 M Bis-Tris propane, pH 7, 20 mM sodium/potassium phosphate, 1% v/v glycerol), crystals appeared after five days, were harvested in mother liquor + 20% v/v glycerol + 10 mM alpha-L-fucose for ~1 minute, then flash-cooled