title: Rescue of syringyl lignin and sinapate ester biosynthesis in Arabidopsis thaliana by a coniferaldehyde 5-hydroxylase from Eucalyptus globulus.
abstract: The gene coding for F5H from Eucalyptus globulus was cloned and used to transform an f5h -mutant of Arabidopsis thaliana , which was complemented, thus verifying the identity of the cloned gene. Coniferaldehyde 5-hydroxylase (F5H; EC 1.14.13) is a cytochrome P450-dependent monooxygenase that catalyzes the 5-hydroxylation step required for the production of syringyl units in lignin biosynthesis. The Eucalyptus globulus enzyme was characterized in vitro, and results showed that the preferred substrates were coniferaldehyde and coniferyl alcohol. Complementation experiments demonstrated that both cDNA and genomic constructs derived from F5H from E. globulus under the control of the cinnamate 4-hydroxylase promoter from Arabidopsis thaliana, or a partial F5H promoter from E. globulus, can rescue the inability of the A. thaliana fah1-2 mutant to accumulate sinapate esters and syringyl lignin. E. globulus is a species widely used to obtain products that require lignin removal, and the results suggest that EglF5H is a good candidate for engineering efforts aimed at increasing the lignin syringyl unit content, either for kraft pulping or biofuel production.
authors.affiliation: Centro de Biotecnología y Facultad Ciencias Forestales, Universidad de Concepción, Concepción, Chile.
authors.forename: José Renán
authors.initials: JR
authors.lastname: García
authors.forename: Nickolas
authors.initials: N
authors.lastname: Anderson
authors.forename: Regis
authors.initials: R
authors.lastname: Le-Feuvre
authors.forename: Carolina
authors.initials: C
authors.lastname: Iturra
authors.forename: Juan
authors.initials: J
authors.lastname: Elissetche
authors.forename: Clint
authors.initials: C
authors.lastname: Chapple
authors.forename: Sofía
authors.initials: S
authors.lastname: Valenzuela
mesh_terms: D000171:Acrolein / Q000031:analogs & derivatives* / Q000378:metabolism; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D005052:Eucalyptus / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D007700:Kinetics; D008031:Lignin / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D029681:Arabidopsis Proteins; D010940:Plant Proteins; C075384:coniferaldehyde; D000171:Acrolein; D008031:Lignin; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C097486:CYP84A1 protein, Arabidopsis
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