Journal: The Journal of Biological Chemistry

Article Title: Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway *

doi: 10.1074/jbc.M113.536250

Figure Lengend Snippet: The proposed β-etherase pathway in Sphingobium sp. strain SYK-6. Panels ( A ) and ( B ) show pathways for metabolism of β( R )- and β( S )-configured β-aryl ether model compounds, with Lig dehydrogenases (LigD, LigL, LigN, and LigO) catalyzing oxidation of the benzylic alcohol in model substrates β( R )- and β( S )-guaiacyl-α-veratrylglycerol, producing β( R )GVG and β( S )GVG. In the presence of GSH, βGVG enantiomers are cleaved to guaiacol and one of two GS-conjugated β-epimers, GS-β( S )VG or GS-β( R )VG, by either LigE, LigP, or LigF. Subsequent GSH-dependent cleavage of GS-β( R )VG by LigG generates β-deoxy-α-veratrylglycerone and GS-SG. C shows the reaction products of Lig enzymes when the achiral model compound βGVE is used as a substrate. In this case, the product of β-etherase activity is GS-βVE, and the product of GS-βVE thioether cleavage by LigG is α-veratrylethanone.

Article Snippet: The LigE and LigP reaction products eluted from the Biotage KP-C18 column with t R = 30.9 and t R = 31.0 min, respectively.

Techniques: Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway *

doi: 10.1074/jbc.M113.536250

Figure Lengend Snippet: HPLC chromatogram traces of pre-enzyme addition (black), LigE ( red ), LigP ( orange ), and LigF ( blue ) enzyme assay samples. A , C 18 -reversed phase chromatography of βGVE prior to the addition of and after a 1-h incubation with GSH and either LigE, LigP, or LigF. B , chiral chromatography of racem -βGVG prior to the addition of and after a 1-h incubation with GSH and either LigE, LigP, or LigF.

Article Snippet: The LigE and LigP reaction products eluted from the Biotage KP-C18 column with t R = 30.9 and t R = 31.0 min, respectively.

Techniques: Enzymatic Assay, Reversed-phase Chromatography, Incubation, Chiral Chromatography

Journal: The Journal of Biological Chemistry

Article Title: Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway *

doi: 10.1074/jbc.M113.536250

Figure Lengend Snippet: Aligned 1 H NMR partial spectra (2.60–4.65 ppm) of the GS conjugate, GS-βVE, in D 2 O. Proton assignment labels correspond with the carbon to which the proton is bound. Alphabetical subscripts differentiate two non-identical geminal protons. Proton peaks that did not integrate as expected are denoted with an asterisk (see “Results”). A , compound GS-βVE obtained via chemical synthesis. B–D , reaction product GS-βVE isolated from LigE ( B ), LigP ( C ), or LigF ( D ) activities using the achiral model compound GβVE as a substrate.

Article Snippet: The LigE and LigP reaction products eluted from the Biotage KP-C18 column with t R = 30.9 and t R = 31.0 min, respectively.

Techniques: Isolation

Journal: The Journal of Biological Chemistry

Article Title: Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway *

doi: 10.1074/jbc.M113.536250

Figure Lengend Snippet: Aligned 1 H NMR partial spectra (2.60–4.65 ppm) of both β-epimers of β- S -glutathionyl-α-veratrylglycerone (GS-β( S )VG and GS-β( R )VG) and both β-epimers of γ,γ-dimethoxy-β- S -(methyl N -acetyl-cysteinyl)-α-veratrylpropanone (CS-β( S )VP and CS-β( R )VP) in D 2 O. Proton assignment labels correspond with the carbon to which the proton is bound. Alphabetical subscripts differentiate two non-identical geminal protons or methoxyls. Red labels and shading are used for the two β( S )-configured compounds, GS-β( S )VG and CS-β( S )VP; blue labels and shading are used for the two β( R )-configured compounds, GS-β( R )VG and CS-β( R )VP; black labels are used for overlapped regions. Regions that differentiate β( S )- and β( R )-configurations ( i.e. protons at β, 2′, and 3′) are shaded across panels. A , chemically synthesized mixture (1:1) of GS-β( S )VG and GS-β( R )VG. B , GS-β( S )VG from LigE. C , GS-β( S )VG from LigP. D , GS-β( R )VG from LigF. E , residual GS-β( S )VG not degraded by LigG. F , chemically synthesized CS-β( S )VP. G , Chemically synthesized CS-β( R )VP.

Article Snippet: The LigE and LigP reaction products eluted from the Biotage KP-C18 column with t R = 30.9 and t R = 31.0 min, respectively.

Techniques: Synthesized