Structure and function of a lignostilbene-α,β-dioxygenase orthologue from Pseudomonas brassicacearum
Open Access
- 16 August 2018
- journal article
- research article
- Published by Springer Science and Business Media LLC in BMC Biochemistry
- Vol. 19 (1), 8
- https://doi.org/10.1186/s12858-018-0098-4
Abstract
Stilbene cleaving oxygenases (SCOs), also known as lignostilbene-α,β-dioxygenases (LSDs) mediate the oxidative cleavage of the olefinic double bonds of lignin-derived intermediate phenolic stilbenes, yielding small modified benzaldehyde compounds. SCOs represent one branch of the larger carotenoid cleavage oxygenases family. Here, we describe the structural and functional characterization of an SCO-like enzyme from the soil-born, bio-control agent Pseudomonas brassicacearum. In vitro and in vivo assays relying on visual inspection, spectrophotometric quantification, as well as liquid-chormatographic and mass spectrometric characterization were applied for functional evaluation of the enzyme. X-ray crystallographic analyses and in silico modeling were applied for structural investigations. In vitro assays demonstrated preferential cleavage of resveratrol, while in vivo analyses detected putative cleavage of the straight chain carotenoid, lycopene. A high-resolution structure containing the seven-bladed β-propeller fold and conserved 4-His-Fe unit at the catalytic site, was obtained. Comparative structural alignments, as well as in silico modelling and docking, highlight potential molecular factors contributing to both the primary in vitro activity against resveratrol, as well as the putative subsidiary activities against carotenoids in vivo, for future validation. The findings reported here provide validation of the SCO structure, and highlight enigmatic points with respect to the potential effect of the enzyme’s molecular environment on substrate specificities for future investigation.Keywords
Funding Information
- National Research Council of Canada (007847)
- Natural Sciences and Engineering Research Council of Canada (356025-2013, 9600-2012, 261683-2012)
- Canadian research chair (Structural Biology, Protein Chemistry)
This publication has 42 references indexed in Scilit:
- MEGA5: Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony MethodsMolecular Biology and Evolution, 2011
- Overview of theCCP4 suite and current developmentsActa crystallographica. Section D, Structural biology, 2011
- Structural Insights into Maize Viviparous14, a Key Enzyme in the Biosynthesis of the Phytohormone Abscisic AcidTHE PLANT CELL ONLINE, 2010
- Features and development of CootActa crystallographica. Section D, Structural biology, 2010
- XDSActa crystallographica. Section D, Structural biology, 2010
- Crystal structure of native RPE65, the retinoid isomerase of the visual cycleProceedings of the National Academy of Sciences of the United States of America, 2009
- Identification of Bacterial Carotenoid Cleavage Dioxygenase Homologues That Cleave the Interphenyl α,β Double Bond of Stilbene Derivatives via a Monooxygenase ReactionChemBioChem, 2008
- Production of a recombinant carotenoid cleavage dioxygenase from grape and enzyme assay in water-miscible organic solventsBiotechnology Letters, 2007
- Refinement of Macromolecular Structures by the Maximum-Likelihood MethodActa crystallographica. Section D, Structural biology, 1997
- Molecular structure and enzymatic function of lycopene cyclase from the cyanobacterium Synechococcus sp strain PCC7942.THE PLANT CELL ONLINE, 1994