1UD2

Crystal structure of calcium-free alpha-amylase from Bacillus sp. strain KSM-K38 (AmyK38)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SPRING-8 BEAMLINE BL45PX
Synchrotron site	SPring-8
Beamline	BL45PX
Temperature [K]	100
Detector technology	IMAGE PLATE
Detector	RIGAKU RAXIS V
Wavelength(s)	1.0200
Spacegroup name	P 2 3
Unit cell lengths	132.143, 132.143, 132.143
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	100.000 * - 2.130
Rwork	0.196
R-free	0.23200 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bli
RMSD bond length	0.006
RMSD bond angle	23.300 *
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	100.000	2.170
High resolution limit [Å]	2.130	2.130
Rmerge	0.105	0.288
Total number of observations	783382 *
Number of reflections	42444 *
<I/σ(I)>	23.5	6.8
Completeness [%]	98.1 *	95.6 *
Redundancy	18.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.5 *	293	PEG 8000, sodium acetate, glycerol, sodium cacodylate, Tris-HCl, pH 6.8, VAPOR DIFFUSION, SITTING DROP, temperature 293K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	Tris-HCl	10 (mM)	pH7.5
2	1	drop	protein	10 (mg/ml)
3	1	reservoir	cacodylate-NaOH	0.085 (M)	pH6.8
4	1	reservoir	PEG8000	25.5 (%(w/v))
5	1	reservoir	sodium acetate	0.17 (M)
6	1	reservoir	glycerol	15 (%(v/v))