6OR3

Structure of an Acyl Intermediate of Thermomyces Lanuginosa Lipase With Palmitic Acid in an Orthorhombic Crystal

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.3.1
Synchrotron site	ALS
Beamline	8.3.1
Temperature [K]	173
Detector technology	PIXEL
Collection date	2019-03-23
Detector	DECTRIS PILATUS 300K
Wavelength(s)	1.1158
Spacegroup name	I 2 2 2
Unit cell lengths	69.865, 85.268, 86.741
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	60.000 - 1.450
R-factor	0.16355
Rwork	0.161
R-free	0.21143
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1tib
RMSD bond length	0.015
RMSD bond angle	1.688
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	REFMAC (5.8.0155)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	60.800	1.470
High resolution limit [Å]	1.450	1.450
Rmerge	0.334	3.600
Rmeas	0.337	3.700
Number of reflections	46198	2213
<I/σ(I)>	10.4	0.6
Completeness [%]	99.9	97.8
Redundancy	62.8	38.7
CC(1/2)	0.997	0.294

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	298	Using filtered culture media with active fungal cells as the stock protein solution, about 30 mg/ml. Sitting drop equal amounts of the protein stock and the reservoir which was 20% PEG 335 buffered with 0.10 M MES at pH 6.5