1GK1

Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	ENRAF-NONIUS FR571
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1997-02-15
Detector	MAR scanner 300 mm plate
Spacegroup name	C 1 2 1
Unit cell lengths	230.290, 70.440, 114.800
Unit cell angles	90.00, 97.48, 90.00

Refinement procedure

Resolution	15.000 * - 2.400
R-factor	0.18
Rwork	0.180
R-free	0.21900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1gk0
RMSD bond length	0.006
RMSD bond angle	24.000 *
Data reduction software	XDS
Data scaling software	XDS
Phasing software	CNS
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	15.000 *	2.500
High resolution limit [Å]	2.400	2.400
Rmerge	0.093 *	0.262
Number of reflections	67222
Completeness [%]	93.7 *	92.7
Redundancy	1.9	1.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5 *	18 *	CRYSTALS GROWN AT 18 C, HANGING DROP PRECIPITATION AGENT: 1.5-2.0 M POTASSIUM PHOSPHATE PH 7.0/9.0

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	20-30 (mg/ml)
2	1	drop	potassium phosphate	0.5 (M)	pH7.5
3	1	drop	dithiothreitol	5 (mM)
4	1	reservoir	potassium phosphate	1.5-2.0	pH7.0-9.0