Literature summary for 1.5.3.5 extracted from

Kachalova, G.S.; Bourenkov, G.P.; Mengesdorf, T.; Schenk, S.; Maun, H.R.; Burghammer, M.; Riekel, C.; Decker, K.; Bartunik, H.D.
Crystal structure analysis of free and substrate-bound 6-hydroxy-L-nicotine oxidase from Arthrobacter nicotinovorans (2010), J. Mol. Biol., 396, 785-799.

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Paenarthrobacter nicotinovorans

Crystallization (Commentary)

Crystallization (Comment)	Organism
His-tagged and untagged free enzyme and complex of dithionite-reduced 6HLNO with the natural substrate 6-hydroxy-L-nicotine X-ray diffraction structure determination and analysis at 1.95 A and 2.05 A resolution, respectively, combined isomorphous/multiple-wavelength anomalous dispersion phasing	Paenarthrobacter nicotinovorans
to 1.95 A resolution. A diacylglycerophospholipid molecule is non-covalently bound to each protomer of 6HLNO. The fatty acid chains occupy hydrophobic channels that penetrate deep into the interior of the substrate-binding domain of each subunit. The solvent-exposed glycerophosphate moiety is located at the subunit-subunit interface. In the crystal structure of a complex of dithionite-reduced 6HLNO with the natural substrate 6-hydroxy-L-nicotine at 2.05 A resolution, the location of the substrate in a tight cavity suggests that the binding geometry of this unproductive complex may be closely similar as under oxidizing conditions. A comparison of the substrate-binding modes of 6HLNO and 6-hydroxy-D-nicotine oxidase, EC 1.5.3.6, based on models of complexes with the D-substrate, suggests that the two enzymes orient the enantiomeric substrates in mirror symmetry with respect to the plane of the flavin	Paenarthrobacter nicotinovorans

Crystallization (Comment)

Organism

His-tagged and untagged free enzyme and complex of dithionite-reduced 6HLNO with the natural substrate 6-hydroxy-L-nicotine X-ray diffraction structure determination and analysis at 1.95 A and 2.05 A resolution, respectively, combined isomorphous/multiple-wavelength anomalous dispersion phasing

Paenarthrobacter nicotinovorans

to 1.95 A resolution. A diacylglycerophospholipid molecule is non-covalently bound to each protomer of 6HLNO. The fatty acid chains occupy hydrophobic channels that penetrate deep into the interior of the substrate-binding domain of each subunit. The solvent-exposed glycerophosphate moiety is located at the subunit-subunit interface. In the crystal structure of a complex of dithionite-reduced 6HLNO with the natural substrate 6-hydroxy-L-nicotine at 2.05 A resolution, the location of the substrate in a tight cavity suggests that the binding geometry of this unproductive complex may be closely similar as under oxidizing conditions. A comparison of the substrate-binding modes of 6HLNO and 6-hydroxy-D-nicotine oxidase, EC 1.5.3.6, based on models of complexes with the D-substrate, suggests that the two enzymes orient the enantiomeric substrates in mirror symmetry with respect to the plane of the flavin

Paenarthrobacter nicotinovorans

Inhibitors

Inhibitors	Comment	Organism	Structure
(R)-6-hydroxynicotine	-	Paenarthrobacter nicotinovorans

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.02	-	(S)-6-hydroxynicotine	pH and temperature not specified in the publication	Paenarthrobacter nicotinovorans

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
(S)-6-hydroxynicotine + H2O + O2	Paenarthrobacter nicotinovorans	absolute stereospecificity on the L-form	1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2	-	?

Organism

Organism	UniProt	Comment	Textmining
Paenarthrobacter nicotinovorans	Q93NH4	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
lipoprotein	a diacylglycerophospholipid molecule is non-covalently bound to each enzyme protomer. The fatty acid chains occupy hydrophobic channels that penetrate deep into the interior of the substrate-binding domain of each subunit. The solvent-exposed glycerophosphate moiety is located at the subunit-subunit interface	Paenarthrobacter nicotinovorans

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(S)-6-hydroxynicotine + H2O + O2	absolute stereospecificity on the L-form	Paenarthrobacter nicotinovorans	1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2	-	?
(S)-6-hydroxynicotine + H2O + O2	absolute stereospecificity on the L-form	Paenarthrobacter nicotinovorans	1-(6-hydroxypyridin-3-yl)-4-(methylamino)butan-1-one + H2O2	intermediate product 6-hydroxy-N-methylmyosmine, which hydrolyzes to 6-hydroxy-pseudooxynicotine	?

Subunits

Subunits	Comment	Organism
dimer	the active enzyme exists as a stable dimer in solution	Paenarthrobacter nicotinovorans

Synonyms

Synonyms	Comment	Organism
6-hydroxy-L-nicotine oxidase	-	Paenarthrobacter nicotinovorans
6HLNO	-	Paenarthrobacter nicotinovorans

Cofactor

Cofactor	Comment	Organism	Structure
FAD	binding domain structure, overview. The flavin may have a role in oxygen activation involving replacement of the water molecule by oxygen and superoxide formation	Paenarthrobacter nicotinovorans

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.1	-	(R)-6-hydroxynicotine	pH and temperature not specified in the publication	Paenarthrobacter nicotinovorans

General Information

General Information	Comment	Organism
additional information	the flavin may have a role in oxygen activation involving replacement of the water molecule by oxygen and superoxide formation. The orientation of the bound substrate relative to the isoalloxazine ring of the FAD cofactor is suitable for hydride transfer dehydrogenation at the carbon atom that forms the chiral center of the substrate molecule, substrate-binding mode, overview. In the dithionite-reduced 6HLNO, the natural substrate 6-hydroxy-L-nicotine is located in a tight cavity suggesting that the binding geometry of this unproductive complex may be closely similar as under oxidizing conditions	Paenarthrobacter nicotinovorans