Journal article
Recombinant Pinoresinol-Lariciresinol Reductases from Western Red Cedar (Thuja plicata) Catalyze Opposite Enantiospecific Conversions
The Journal of biological chemistry, Vol.274(2), pp.618-627
01/08/1999
Handle:
https://hdl.handle.net/2376/105817
PMID: 9872995
Abstract
Although the heartwood of woody plants represents the main source of fiber and solid wood products, essentially nothing is known about how the biological processes leading to its formation are initiated and regulated. Accordingly, a reverse transcription-polymerase chain reaction-guided cloning strategy was employed to obtain genes encoding pinoresinol-lariciresinol reductases from western red cedar (Thuja plicata) as a means to initiate the study of its heartwood formation. (+)-Pinoresinol-(+)-lariciresinol reductase from Forsythia intermedia was used as a template for primer construction for reverse transcription-polymerase chain reaction amplifications, which, when followed by homologous hybridization cloning, resulted in the isolation of two distinct classes of putative pinoresinol-lariciresinol reductase cDNA clones from western red cedar. A representative of each class was expressed as a fusion protein with β-galactosidase and assayed for enzymatic activity. Using both deuterated and radiolabeled (±)-pinoresinols as substrates, it was established that each class of cDNA encoded a pinoresinol-lariciresinol reductase of different (opposite) enantiospecificity. Significantly, the protein from one class converted (+)-pinoresinol into (–)-secoisolariciresinol, whereas the other utilized the opposite (–)-enantiomer to give the corresponding (+)-form. This differential substrate specificity raises important questions about the role of each of these individual reductases in heartwood formation, such as whether they are expressed in different cells/tissues or at different stages during heartwood development.
Metrics
7 Record Views
Details
- Title
- Recombinant Pinoresinol-Lariciresinol Reductases from Western Red Cedar (Thuja plicata) Catalyze Opposite Enantiospecific Conversions
- Creators
- Masayuki Fujita - Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 and theDavid R Gang - Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 and theLaurence B Davin - Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 and theNorman G Lewis - Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340 and the
- Publication Details
- The Journal of biological chemistry, Vol.274(2), pp.618-627
- Academic Unit
- Biological Chemistry, Institute of
- Publisher
- Elsevier Inc
- Identifiers
- 99900546851201842
- Language
- English
- Resource Type
- Journal article