8RIO

Beta-keto acid cleavage enzyme from Paracoccus denitrificans

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P14 (MX2)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P14 (MX2)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-08-20
Detector	DECTRIS EIGER2 X CdTe 16M
Wavelength(s)	0.9763
Spacegroup name	C 2 2 21
Unit cell lengths	81.065, 137.871, 132.517
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.780 - 1.450
R-factor	0.1567
Rwork	0.156
R-free	0.17680
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.015
RMSD bond angle	1.425
Data reduction software	XDS (20210323)
Data scaling software	SCALA (3.3.22)
Phasing software	PHENIX (1.20.1_4487)
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	24.780	1.530
High resolution limit [Å]	1.450	1.450
Rmerge	0.090	0.499
Rmeas	0.093	0.525
Rpim	0.026	0.157
Total number of observations	1572792	194961
Number of reflections	129609	18038
<I/σ(I)>	19.2	5.9
Completeness [%]	99.1
Redundancy	12.1	10.8
CC(1/2)	0.997	0.929

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	The enzyme (8.6 mg/mL) in 50 mM HEPES pH 7.8, 150 mM KCl, 1 M L-proline, and 1 mM ZnCl2 was mixed in a 1:1 ratio with 25 % (w/v) pentaerythritol propoxylate (17/8 PO/OH), 100 mM HEPES pH 7.5. The final size of the drops was 1 microliter. Prior to flash freezing the crystals in liquid nitrogen, the mother liquor was supplemented with 37 % (w/v) pentaerythrol propoxylate (17/8 PO/OH).