Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway
Abstract
Glutathione-dependent enzymes play important protective, repair, or metabolic roles in cells. In particular, enzymes in the glutathione S-transferase (GST) superfamily function in stress responses, defense systems, or xenobiotic detoxification. Here, we identify novel features of bacterial GSTs that cleave -aryl ether bonds typically found in plant lignin. Our data reveal several original features of the reaction cycle of these GSTs, including stereospecific substrate recognition and stereoselective formation of -S-thioether linkages. Products of recombinant GSTs (LigE, LigP, and LigF) are -S-glutathionyl--ketothioethers that are degraded by a -S-thioetherase (LigG). All three Lig GSTs produced the ketone product (-S-glutathionyl- -veratrylethanone) from an achiral side chain-truncated model substrate (-guaiacyl--veratrylethanone). However, when -etherase assays were conducted with a racemic model substrate, -guaiacyl--veratrylglycerone, LigE- or LigP-catalyzed reactions yielded only one of two potential product (-Sglutathionyl--veratrylglycerone) epimers, whereas the other diastereomer (differing in configuration at the -position (i.e. its-epimer)) was produced only in the LigF-catalyzed reaction. Thus, -etherase catalysis causes stereochemical inversion of the chiral center, converting a (R)-substrate to a (S)-product (LigE and LigP), and a (S)-substrate to a (R)-product (LigF). Further, LigG catalyzed glutathione-dependent -S-thioether cleavage with-S-glutathionyl--veratrylethanone and with(R)- configured -S-glutathionyl--veratrylglycerone but exhibited no or significantly reduced -S-thioether-cleaving activity with the (S)-epimer, demonstrating that LigGmore »
- Authors:
-
- University of Wisconsin, Madison, WI (United States)
- Publication Date:
- Research Org.:
- Univ. of Wisconsin, Madison, WI (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1904221
- Grant/Contract Number:
- FC02-07ER64494
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Biological Chemistry
- Additional Journal Information:
- Journal Volume: 289; Journal Issue: 12; Journal ID: ISSN 0021-9258
- Publisher:
- American Society for Biochemistry and Molecular Biology
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; bacterial metabolism; enzyme catalysis; glutathione; lignin degradation; thiol; beta-S-thioetherase; beta-aryl etherase; glutathione s-transferase; stereoselectivity; stereospecificity
Citation Formats
Gall, Daniel L., Kim, Hoon, Lu, Fachuang, Donohue, Timothy J., Noguera, Daniel R., and Ralph, John. Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway. United States: N. p., 2014.
Web. doi:10.1074/jbc.m113.536250.
Gall, Daniel L., Kim, Hoon, Lu, Fachuang, Donohue, Timothy J., Noguera, Daniel R., & Ralph, John. Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway. United States. https://doi.org/10.1074/jbc.m113.536250
Gall, Daniel L., Kim, Hoon, Lu, Fachuang, Donohue, Timothy J., Noguera, Daniel R., and Ralph, John. Fri .
"Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway". United States. https://doi.org/10.1074/jbc.m113.536250. https://www.osti.gov/servlets/purl/1904221.
@article{osti_1904221,
title = {Stereochemical Features of Glutathione-dependent Enzymes in the Sphingobium sp. Strain SYK-6 β-Aryl Etherase Pathway},
author = {Gall, Daniel L. and Kim, Hoon and Lu, Fachuang and Donohue, Timothy J. and Noguera, Daniel R. and Ralph, John},
abstractNote = {Glutathione-dependent enzymes play important protective, repair, or metabolic roles in cells. In particular, enzymes in the glutathione S-transferase (GST) superfamily function in stress responses, defense systems, or xenobiotic detoxification. Here, we identify novel features of bacterial GSTs that cleave -aryl ether bonds typically found in plant lignin. Our data reveal several original features of the reaction cycle of these GSTs, including stereospecific substrate recognition and stereoselective formation of -S-thioether linkages. Products of recombinant GSTs (LigE, LigP, and LigF) are -S-glutathionyl--ketothioethers that are degraded by a -S-thioetherase (LigG). All three Lig GSTs produced the ketone product (-S-glutathionyl- -veratrylethanone) from an achiral side chain-truncated model substrate (-guaiacyl--veratrylethanone). However, when -etherase assays were conducted with a racemic model substrate, -guaiacyl--veratrylglycerone, LigE- or LigP-catalyzed reactions yielded only one of two potential product (-Sglutathionyl--veratrylglycerone) epimers, whereas the other diastereomer (differing in configuration at the -position (i.e. its-epimer)) was produced only in the LigF-catalyzed reaction. Thus, -etherase catalysis causes stereochemical inversion of the chiral center, converting a (R)-substrate to a (S)-product (LigE and LigP), and a (S)-substrate to a (R)-product (LigF). Further, LigG catalyzed glutathione-dependent -S-thioether cleavage with-S-glutathionyl--veratrylethanone and with(R)- configured -S-glutathionyl--veratrylglycerone but exhibited no or significantly reduced -S-thioether-cleaving activity with the (S)-epimer, demonstrating that LigG is a stereospecific -thioetherase. We therefore propose that multiple Lig enzymes are needed in this -aryl etherase pathway in order to cleave the racemic -ether linkages that are present in the backbone of the lignin polymer.},
doi = {10.1074/jbc.m113.536250},
journal = {Journal of Biological Chemistry},
number = 12,
volume = 289,
place = {United States},
year = {Fri Feb 07 00:00:00 EST 2014},
month = {Fri Feb 07 00:00:00 EST 2014}
}
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