3D2C

Structure of 4D3, 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	C 2 2 21
Unit cell lengths	132.285, 198.042, 197.902
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	25.000 - 2.180
Rwork	0.202
R-free	0.24500
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	2.260
High resolution limit [Å]	2.180	4.690	2.180
Rmerge	0.096	0.040	0.489
Number of reflections	133252
<I/σ(I)>	8.5
Completeness [%]	98.6	100	93.6
Redundancy	4.1	4.5	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	298	1M succinic acid, 0.1M HEPES, 1%(w/v) polyethelene glycol monomethyl ether 2000, pH 7.0, VAPOR DIFFUSION, SITTING DROP, temperature 298K