3D2B

Structure of 2D9, a thermostable mutant of Bacillus subtilis lipase obtained through directed evolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RUH3R
Temperature [K]	100
Detector technology	IMAGE PLATE
Detector	MAR scanner 345 mm plate
Wavelength(s)	1.5418
Spacegroup name	I 2 2 2
Unit cell lengths	75.203, 117.399, 117.555
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	25.000 - 1.950
Rwork	0.184
R-free	0.21000
Structure solution method	MOLECULAR REPLACEMENT
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	CNS

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	25.000	25.000	2.020
High resolution limit [Å]	1.950	4.200	1.950
Rmerge	0.066	0.026	0.277
Number of reflections	36223
<I/σ(I)>	10.2
Completeness [%]	94.3	86.5	93.2
Redundancy	3.6	3.9	3.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9.5	277	100mM Ethanolamine buffe, 10mM Sodium sulfate, pH 9.5, vapour diffusion, sitting drop, temperature 277K, VAPOR DIFFUSION, SITTING DROP