6K98

Substrates promiscuity of xyloglucanases and endoglucanases of glycoside hydrolase 12 family

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSRRC BEAMLINE BL15A1
Synchrotron site	NSRRC
Beamline	BL15A1
Temperature [K]	298
Detector technology	PIXEL
Collection date	2019-01-04
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.00919
Spacegroup name	P 43 21 2
Unit cell lengths	92.708, 92.708, 119.928
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	57.522 - 2.032
R-factor	0.1866
Rwork	0.185
R-free	0.21200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ks4
RMSD bond length	0.007
RMSD bond angle	0.839
Phasing software	PHASER
Refinement software	PHENIX ((1.14_3260: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	57.522	4.519
High resolution limit [Å]	2.032	2.033
Number of reflections	29949	11
<I/σ(I)>	1.35
Completeness [%]	87.2
Redundancy	4.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		298.15	NfEG12A was concentrated to 10 mg/ml in a buffer containing 100 mM citric acid-Na2HPO4 (pH 7.2). The protein was crystallized in a mother liquid with 0.1 M citrate (pH 5.0) and 16% (w/v) PEG 20000 using the hanging drop vapor diffusion method in 24-well plates. NfEG12A crystals was transferred to a reservoir solution supplemented with 25% (v/v) ethylene glycol to grown. Obtained crystals were cryoprotected with 25% (v/v) 2-methyl-2,4-pentanediol. The crystals were mounted on a nylon loop and cooled immediately in liquid nitrogen prior to data collection.