Crystal Structure of BoxB in mixed valent state with bound benzoyl-CoA

Structure and Mechanism of the Diiron Benzoyl-Coenzyme A Epoxidase BoxB.

LN2 cooled fixed-exit
Si(111) monochromator

Benzoyl-CoA 2,3-dioxygenase subunit B, Benzoyl-CoA dioxygenase oxygenase component

MINYSERIPNNVNLNENKTLQRALEQWQPSFLNWWDDMGPENSSNYDVYLRTAVSVDPKGWADFGYVKMHDYRWGIFLAP
QEGEKKITFGEHKGQDVWQEVPGEYRSTLRRIIVTQGDTEPASVEQQRHLGLTAPSLYDLRNLFQVNVEEGRHLWAMVYL
LHAHFGRDGREEGEALLERRSGDEDNPRILTAFNEKTPDWLSFFMFTFITDRDGKFQLASLAESAFDPLARTCKFMLTEE
AHHLFVGESGIARVIQRTCEVMKELGTDDPAKLRAAGVIDLPTLQKYLNFHYSVTSDLYGAEISSNAATYYTNGLKGRFE
EEKIGDDHKLQNSEYEVMDVAGDKILTRHVPALSALNERLRDDWITDVQAGVDRWNRIPAKFGFDFRFTLPHKGFHRKIG
MFADVHVSPDGRLISEAEWTHQHKNWLPTESDRLYVHSLMGRCLEPGKFANWIAAPARGINNQPVNFEYVRFNWSHPQFE
K

10% PEG 4000, 0.1M sodium acetate, 0.1M Li2SO4, 18% glycerol, 5 mM benzoyl-CoA, 1 mM NADPH, 0.1 mg/ml BoxA 10 mM Pipes, 0.1 M KCl, VAPOR DIFFUSION, HANGING DROP, temperature 298K, pH 4.6

THE C-TERMINAL TAGGED BOXB WAS INTEGRATED INTO THE GENOME OF AZOARCUS EVANSII AS DESCRIBED ELSEWHERE FOR THE HIS-TAG VERSION (ZAAR ET AL. 2004)

HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS

diiron center, epoxidase, oxidoreductase, benzoyl coenzyme A

MINYSERIPNNVNLNENKTLQRALEQWQPSFLNWWDDMGPENSSNYDVYLRTAVSVDPKGWADFGYVKMHDYRWGIFLAP
QEGEKKITFGEHKGQDVWQEVPGEYRSTLRRIIVTQGDTEPASVEQQRHLGLTAPSLYDLRNLFQVNVEEGRHLWAMVYL
LHAHFGRDGREEGEALLERRSGDEDNPRILTAFNEKTPDWLSFFMFTFITDRDGKFQLASLAESAFDPLARTCKFMLTEE
AHHLFVGESGIARVIQRTCEVMKELGTDDPAKLRAAGVIDLPTLQKYLNFHYSVTSDLYGAEISSNAATYYTNGLKGRFE
EEKIGDDHKLQNSEYEVMDVAGDKILTRHVPALSALNERLRDDWITDVQAGVDRWNRIPAKFGFDFRFTLPHKGFHRKIG
MFADVHVSPDGRLISEAEWTHQHKNWLPTESDRLYVHSLMGRCLEPGKFANWIAAPARGINNQPVNFEYVRFN