1OB0

Kinetic stabilization of Bacillus licheniformis alpha-amylase through introduction of hydrophobic residues at the surface

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	278
Detector technology	IMAGE PLATE
Collection date	1997-05-15
Detector	MARRESEARCH
Spacegroup name	P 61
Unit cell lengths	91.292, 91.292, 137.466
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	25.890 - 1.830
R-factor	0.147
Rwork	0.147
R-free	0.17400 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bpl
RMSD bond length	0.009 *
RMSD bond angle	1.359 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.900	1.850
High resolution limit [Å]	1.830	1.700 *
Rmerge	0.073 *	0.310 *
Total number of observations	231592 *
Number of reflections	61355 *
<I/σ(I)>	19	2.1
Completeness [%]	83.8 *	25 *
Redundancy	3.8 *	2.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	8 *	20 *	PROTEIN WAS CRYSTALLIZED BY VAPOR DIFFUSION FROM DROPS CONTAINING 4 UL OF PROTEIN SOLUTION (10 MG/ML IN 50 MM TRIS/HCL, PH 8.0) PLUS 4 UL OF RESERVOIR SOLUTION (50 MM HEPES, 1 M AMMONIUM SULFATE, 1% (V/V) PEG 500, PH 7.0) EQUILIBRATED AGAINST 1 ML OF RESERVOIR SOLUTION.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	10 (mg/ml)
2	1	drop	Tris-HCl	50 (mM)	pH8.0
3	1	reservoir	HEPES	50 (mM)
4	1	reservoir	ammonium sulfate	1 (M)
5	1	reservoir	PEG500	1 (%(v/v))	pH7.0