7QT1

Non-obligately L8S8-complex forming RubisCO derived from ancestral sequence reconstruction and rational engineering in L8S8 complex with substitution e170N

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-2
Synchrotron site	ESRF
Beamline	ID23-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-12-02
Detector	DECTRIS PILATUS3 2M
Wavelength(s)	0.873128
Spacegroup name	I 1 2 1
Unit cell lengths	108.942, 106.604, 191.968
Unit cell angles	90.00, 98.78, 90.00

Refinement procedure

Resolution	29.290 - 2.100
R-factor	0.1633
Rwork	0.163
R-free	0.18570
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6ura
Data reduction software	XDS (20210323)
Data scaling software	SCALA (3.3.22)
Phasing software	PHENIX (1.18.2_3874)
Refinement software	PHENIX (1.18.2_3874)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	29.290	29.286	2.210
High resolution limit [Å]	2.100	6.640	2.100
Rmerge		0.068	0.710
Rmeas	0.239	0.075	0.777
Rpim	0.096	0.031	0.311
Total number of observations		22987	112922
Number of reflections	126281	4004	18399
<I/σ(I)>	7.1	17.7	2.4
Completeness [%]	99.9	97.3	100
Redundancy	6	5.7	6.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9.1	288	Purified enzyme (3.25 mg/mL) in 25 mM Tricine-NaOH, 75 mM NaCl, pH 8.0 was incubated at 3% (v/v) CO2 in air and 30 degrees C for 1 h in the presence of 0.182 mM CABP and 2.9 mM MgCl2. The enzyme was then mixed in a 1:1 ratio with 0.2 M Bis-Tris propane, 20 % (w/v) polyethylene glycol 4000, pH 9.1. Before flash freezing the crystals in liquid nitrogen PEG200 was added to the mother liquor as cryoprotectant to a final concentration of 33 % (w/v).