title: XAX1 from glycosyltransferase family 61 mediates xylosyltransfer to rice xylan.
abstract: Xylan is the second most abundant polysaccharide on Earth and represents an immense quantity of stored energy for biofuel production. Despite its importance, most of the enzymes that synthesize xylan have yet to be identified. Xylans have a backbone of β-1,4-linked xylose residues with substitutions that include α-(1→2)-linked glucuronosyl, 4-O-methyl glucuronosyl, and α-1,2- and α-1,3-arabinofuranosyl residues. The substitutions are structurally diverse and vary by taxonomy, with grass xylan representing a unique composition distinct from dicots and other monocots. To date, no enzyme has yet been identified that is specific to grass xylan synthesis. We identified a xylose-deficient loss-of-function rice mutant in Os02g22380, a putative glycosyltransferase in a grass-specific subfamily of family GT61. We designate the mutant xax1 for xylosyl arabinosyl substitution of xylan 1. Enzymatic fingerprinting of xylan showed the specific absence in the mutant of a peak, which was isolated and determined by (1)H-NMR to be (β-1,4-Xyl)(4) with a β-Xylp-(1→2)-α-Araf-(1→3). Rice xax1 mutant plants are deficient in ferulic and coumaric acid, aromatic compounds known to be attached to arabinosyl residues in xylan substituted with xylosyl residues. The xax1 mutant plants exhibit an increased extractability of xylan and increased saccharification, probably reflecting a lower degree of diferulic cross-links. Activity assays with microsomes isolated from tobacco plants transiently expressing XAX1 demonstrated xylosyltransferase activity onto endogenous acceptors. Our results provide insight into grass xylan synthesis and how substitutions may be modified for increased saccharification for biofuel generation.
authors.affiliation: Department of Plant Pathology and the Genome Center, University of California, Davis, CA 95616, USA.
authors.forename: Dawn
authors.initials: D
authors.lastname: Chiniquy
authors.forename: Vaishali
authors.initials: V
authors.lastname: Sharma
authors.forename: Alex
authors.initials: A
authors.lastname: Schultink
authors.forename: Edward E
authors.initials: EE
authors.lastname: Baidoo
authors.forename: Carsten
authors.initials: C
authors.lastname: Rautengarten
authors.forename: Kun
authors.initials: K
authors.lastname: Cheng
authors.forename: Andrew
authors.initials: A
authors.lastname: Carroll
authors.forename: Peter
authors.initials: P
authors.lastname: Ulvskov
authors.forename: Jesper
authors.initials: J
authors.lastname: Harholt
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
authors.forename: Markus
authors.initials: M
authors.lastname: Pauly
authors.forename: Henrik V
authors.initials: HV
authors.lastname: Scheller
authors.forename: Pamela C
authors.initials: PC
authors.lastname: Ronald
mesh_terms: D056804:Biofuels; D002473:Cell Wall / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008861:Microsomes; D012275:Oryza / Q000201:enzymology* / Q000378:metabolism; D010430:Pentosyltransferases / Q000235:genetics / Q000378:metabolism*; D014990:Xylans / Q000378:metabolism*; D014994:Xylose / Q000378:metabolism*; D000097608:UDP Xylose-Protein Xylosyltransferase
chemical_list: D056804:Biofuels; D010430:Pentosyltransferases; D014990:Xylans; D014994:Xylose; D000097608:UDP Xylose-Protein Xylosyltransferase