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Determinants and Expansion of Specificity in a Trichothecene UDP-Glucosyltransferase from Oryza sativa

Journal Article · · Biochemistry
 [1];  [1];  [2];  [2];  [3];  [3];  [2];  [2];  [1]
  1. Univ. of Wisconsin, Madison, WI (United States)
  2. Univ. of Natural Resources and Life Sciences, Tulln (Austria)
  3. National Center for Agricultural Utilization Research, Peoria, IL (United States)
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Family 1 UDP-glycosyltransferases (UGTs) in plants primarily form glucose conjugates of small molecules and, besides other functions, play a role in detoxification of xenobiotics. Indeed, overexpression of a barley UGT in wheat has been shown to control Fusarium head blight, which is a plant disease of global significance that leads to reduced crop yields and contamination with trichothecene mycotoxins such as deoxynivalenol (DON), T-2 toxin, and many other structural variants. The UGT Os79 from rice has emerged as a promising candidate for inactivation of mycotoxins because of its ability to glycosylate DON, nivalenol, and hydrolyzed T-2 toxin (HT-2). However, Os79 is unable to modify T-2 toxin (T-2), produced by pathogens such as Fusarium sporotrichioides and Fusarium langsethii. Activity toward T-2 is desirable because it would allow a single UGT to inactivate co-occurring mycotoxins. Here, the structure of Os79 in complex with the products UDP and deoxynivalenol 3-O-glucoside is reported together with a kinetic analysis of a broad range of trichothecene mycotoxins. Residues associated with the trichothecene binding pocket were examined by site-directed mutagenesis that revealed that trichothecenes substituted at the C4 position, which are not glycosylated by wild-type Os79, can be accommodated in the binding pocket by increasing its volume. The H122A/L123A/Q202L triple mutation, which increases the volume of the active site and attenuates polar contacts, led to strong and equivalent activity toward trichothecenes with C4 acetyl groups. Lastly, this mutant enzyme provides the broad specificity required to control multiple toxins produced by different Fusarium species and chemotypes.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE; U.S. Dept. of Agriculture; Vienna Science and Technology; Austrian Science Fund
Grant/Contract Number:
W-31109-ENG-38
OSTI ID:
1414464
Journal Information:
Biochemistry, Journal Name: Biochemistry Journal Issue: 50 Vol. 56; ISSN 0006-2960
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
ENGLISH

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Cited By (3)

Enzymes for Detoxification of Various Mycotoxins: Origins and Mechanisms of Catalytic Action journal June 2019
A systematic review of plant-conjugated masked mycotoxins: Occurrence, toxicology, and metabolism journal February 2019
UDP-Glucosyltransferases from Rice, Brachypodium, and Barley: Substrate Specificities and Synthesis of Type A and B Trichothecene-3-O-β-d-glucosides journal March 2018

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