6K9D

glycoside hydrolase family 12 (GH12) englucanase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSRRC BEAMLINE BL15A1
Synchrotron site	NSRRC
Beamline	BL15A1
Temperature [K]	100
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	88.631, 88.631, 123.411
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	55.879 - 1.505
R-factor	0.1738
Rwork	0.173
R-free	0.18620
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.806
Data scaling software	xia2
Phasing software	PHASER
Refinement software	PHENIX ((1.14_3260: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	55.879	4.279
High resolution limit [Å]	1.505	1.505
Number of reflections	62640	23
<I/σ(I)>	1.34
Completeness [%]	80.0
Redundancy	4.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		298.15	NfEG12A-N18Y was concentrated to 10 mg/ml in a buffer containing 100 mM citric acid-Na2HPO4 (pH 7.2). both proteins were 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-N18Y 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.