Isolation and some properties of lysine N6-hydroxylase from Escherichia coli strain EN222.
Lysine N6-hydroxylase was isolated as a soluble enzyme from the supernatant after ultrasonication of Escherichia coli strain EN222 which contained the structural gene on a multicopy plasmid (as described by Engelbrecht and Braun in 1986). The apoenzyme prepared by dialysis was purified by ammonium sulfate precipitation and fast protein liquid chromatography using Superose 12 and Mono Q columns. The molecular mass as determined by gel filtration was 200 kDa and 50 kDa by SDS/polyacrylamide gel electrophoresis. The enzyme binds 0.79 molecule FAD/50 kDa. The activity of the enzyme is strictly dependent on NADPH. Its properties are similar to other flavoprotein monooxygenases of the EC group 1.14.13.
Plattner
H J
HJ
Institut für Mikrobiologie, Universität Hannover, Federal Republic of Germany.
Pfefferle
P
P
Romaguera
A
A
Waschütza
S
S
Diekmann
H
H
D000595:Amino Acid Sequence; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D006901:Hydroxylysine / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000302:isolation & purification* / Q000378:metabolism
D006901:Hydroxylysine; D009249:NADP; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase