29CU

Crystal Structure of a Cutinase from Saccharopolyspora Dendranthemae

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I03
Synchrotron site	Diamond
Beamline	I03
Temperature [K]	100
Detector technology	PIXEL
Collection date	2024-02-02
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.97625
Spacegroup name	P 21 21 21
Unit cell lengths	38.502, 61.707, 88.128
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.550 - 1.130
R-factor	0.12105
Rwork	0.120
R-free	0.14444
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.013
RMSD bond angle	1.834
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	PHASER
Refinement software	REFMAC (5.8.0425)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.550	1.200
High resolution limit [Å]	1.130	1.130
Rmerge	0.057	0.500
Rmeas	0.060	0.650
Number of reflections	67263	443
<I/σ(I)>	21.8	1.4
Completeness [%]	84.1	11.2
Redundancy	11.1
CC(1/2)	0.990	0.650

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277.15	Crystals were grown by sitting drop vapor diffusion at 4 degrees C. Drops were set up by mixing 1 uL of protein solution (10 mg/mL in Y sodium phosphate buffer, pH 7.5) with 1 uL of reservoir solution (0.2 M Sodium Fluoride, 20% (w/v) PEG 3350, unbuffered). The drops were incubated for 9 months. The setup was performed using a TTP Labtech Mosquito liquid handling robot.