UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides
Trichothecene toxins are confirmed or suspected virulence factors of various plant-pathogenic Fusarium species. Plants can detoxify these to a variable extent by glucosylation, a reaction catalyzed by UDP-glucosyltransferases (UGTs). Due to the unavailability of analytical standards for many trichot...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2018-03-01
|
| Series: | Toxins |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2072-6651/10/3/111 |
| _version_ | 1811183952009887744 |
|---|---|
| author | Herbert Michlmayr Elisabeth Varga Alexandra Malachová Philipp Fruhmann Marta Piątkowska Christian Hametner Jana Šofrová Günther Jaunecker Georg Häubl Marc Lemmens Franz Berthiller Gerhard Adam |
| author_facet | Herbert Michlmayr Elisabeth Varga Alexandra Malachová Philipp Fruhmann Marta Piątkowska Christian Hametner Jana Šofrová Günther Jaunecker Georg Häubl Marc Lemmens Franz Berthiller Gerhard Adam |
| author_sort | Herbert Michlmayr |
| collection | DOAJ |
| description | Trichothecene toxins are confirmed or suspected virulence factors of various plant-pathogenic Fusarium species. Plants can detoxify these to a variable extent by glucosylation, a reaction catalyzed by UDP-glucosyltransferases (UGTs). Due to the unavailability of analytical standards for many trichothecene-glucoconjugates, information on such compounds is limited. Here, the previously identified deoxynivalenol-conjugating UGTs HvUGT13248 (barley), OsUGT79 (rice) and Bradi5g03300 (Brachypodium), were expressed in E. coli, affinity purified, and characterized towards their abilities to glucosylate the most relevant type A and B trichothecenes. HvUGT13248, which prefers nivalenol over deoxynivalenol, is also able to conjugate C-4 acetylated trichothecenes (e.g., T-2 toxin) to some degree while OsUGT79 and Bradi5g03300 are completely inactive with C-4 acetylated derivatives. The type A trichothecenes HT-2 toxin and T-2 triol are the kinetically preferred substrates in the case of HvUGT13248 and Bradi5g03300. We glucosylated several trichothecenes with OsUGT79 (HT-2 toxin, T-2 triol) and HvUGT13248 (T-2 toxin, neosolaniol, 4,15-diacetoxyscirpenol, fusarenon X) in the preparative scale. NMR analysis of the purified glucosides showed that exclusively β-D-glucosides were formed regio-selectively at position C-3-OH of the trichothecenes. These synthesized standards can be used to investigate the occurrence and toxicological properties of these modified mycotoxins. |
| first_indexed | 2024-04-11T13:05:17Z |
| format | Article |
| id | doaj.art-b68bdef612f64cef9f03a9e76ae4f832 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T13:05:17Z |
| publishDate | 2018-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-b68bdef612f64cef9f03a9e76ae4f8322022-12-22T04:22:48ZengMDPI AGToxins2072-66512018-03-0110311110.3390/toxins10030111toxins10030111UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosidesHerbert Michlmayr0Elisabeth Varga1Alexandra Malachová2Philipp Fruhmann3Marta Piątkowska4Christian Hametner5Jana Šofrová6Günther Jaunecker7Georg Häubl8Marc Lemmens9Franz Berthiller10Gerhard Adam11Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, (BOKU), Konrad Lorenz Str. 24, 3430 Tulln, AustriaChristian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), BOKU, Konrad Lorenz Str. 20, 3430 Tulln, AustriaChristian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), BOKU, Konrad Lorenz Str. 20, 3430 Tulln, AustriaInstitute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163, 1060 Vienna, AustriaChristian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), BOKU, Konrad Lorenz Str. 20, 3430 Tulln, AustriaInstitute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163, 1060 Vienna, AustriaChristian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), BOKU, Konrad Lorenz Str. 20, 3430 Tulln, AustriaRomerlabs Division Holding GmbH, Technopark 1, 3430 Tulln, AustriaRomerlabs Division Holding GmbH, Technopark 1, 3430 Tulln, AustriaBiotechnology in Plant Production, IFA-Tulln, BOKU, Konrad Lorenz Str. 20, 3430 Tulln, AustriaChristian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), BOKU, Konrad Lorenz Str. 20, 3430 Tulln, AustriaDepartment of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, (BOKU), Konrad Lorenz Str. 24, 3430 Tulln, AustriaTrichothecene toxins are confirmed or suspected virulence factors of various plant-pathogenic Fusarium species. Plants can detoxify these to a variable extent by glucosylation, a reaction catalyzed by UDP-glucosyltransferases (UGTs). Due to the unavailability of analytical standards for many trichothecene-glucoconjugates, information on such compounds is limited. Here, the previously identified deoxynivalenol-conjugating UGTs HvUGT13248 (barley), OsUGT79 (rice) and Bradi5g03300 (Brachypodium), were expressed in E. coli, affinity purified, and characterized towards their abilities to glucosylate the most relevant type A and B trichothecenes. HvUGT13248, which prefers nivalenol over deoxynivalenol, is also able to conjugate C-4 acetylated trichothecenes (e.g., T-2 toxin) to some degree while OsUGT79 and Bradi5g03300 are completely inactive with C-4 acetylated derivatives. The type A trichothecenes HT-2 toxin and T-2 triol are the kinetically preferred substrates in the case of HvUGT13248 and Bradi5g03300. We glucosylated several trichothecenes with OsUGT79 (HT-2 toxin, T-2 triol) and HvUGT13248 (T-2 toxin, neosolaniol, 4,15-diacetoxyscirpenol, fusarenon X) in the preparative scale. NMR analysis of the purified glucosides showed that exclusively β-D-glucosides were formed regio-selectively at position C-3-OH of the trichothecenes. These synthesized standards can be used to investigate the occurrence and toxicological properties of these modified mycotoxins.http://www.mdpi.com/2072-6651/10/3/111Fusarium head blightphase II detoxificationmasked mycotoxinNMRcereals |
| spellingShingle | Herbert Michlmayr Elisabeth Varga Alexandra Malachová Philipp Fruhmann Marta Piątkowska Christian Hametner Jana Šofrová Günther Jaunecker Georg Häubl Marc Lemmens Franz Berthiller Gerhard Adam UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides Toxins Fusarium head blight phase II detoxification masked mycotoxin NMR cereals |
| title | UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides |
| title_full | UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides |
| title_fullStr | UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides |
| title_full_unstemmed | UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides |
| title_short | UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides |
| title_sort | udp glucosyltransferases from rice brachypodium and barley substrate specificities and synthesis of type a and b trichothecene 3 o β d glucosides |
| topic | Fusarium head blight phase II detoxification masked mycotoxin NMR cereals |
| url | http://www.mdpi.com/2072-6651/10/3/111 |
| work_keys_str_mv | AT herbertmichlmayr udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT elisabethvarga udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT alexandramalachova udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT philippfruhmann udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT martapiatkowska udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT christianhametner udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT janasofrova udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT guntherjaunecker udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT georghaubl udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT marclemmens udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT franzberthiller udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides AT gerhardadam udpglucosyltransferasesfromricebrachypodiumandbarleysubstratespecificitiesandsynthesisoftypeaandbtrichothecene3obdglucosides |