XAX1 from glycosyltransferase family 61 mediates xylosyltransfer to rice xylan

Dawn Chiniquy; Vaishali Sharma; Alex Schultink; Edward E. Baidoo; Carsten Rautengarten; Kun Cheng; Andrew Carroll; Peter Ulvskov; Jesper Harholt; Jay D. Keasling; Markus Pauly; Henrik V. Scheller; Pamela C. Ronald

doi:10.1073/pnas.1202079109

XAX1 from glycosyltransferase family 61 mediates xylosyltransfer to rice xylan

Dawn Chiniquy, Vaishali Sharma, Alex Schultink, Edward E. Baidoo, Carsten Rautengarten, Kun Cheng, Andrew Carroll, Peter Ulvskov, Jesper Harholt, Jay D. Keasling, Markus Pauly, Henrik V. Scheller, Pamela C. Ronald

119 Citations (Scopus)

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 residueswith 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 ¹H-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.

Original language	English
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	109
Issue number	42
Pages (from-to)	17117-17122
Number of pages	6
ISSN	0027-8424
DOIs	https://doi.org/10.1073/pnas.1202079109
Publication status	Published - 16 Oct 2012

Access to Document

10.1073/pnas.1202079109

Cite this

Chiniquy, D., Sharma, V., Schultink, A., Baidoo, E. E., Rautengarten, C., Cheng, K., Carroll, A., Ulvskov, P., Harholt, J., Keasling, J. D., Pauly, M., Scheller, H. V., & Ronald, P. C. (2012). XAX1 from glycosyltransferase family 61 mediates xylosyltransfer to rice xylan. Proceedings of the National Academy of Sciences of the United States of America, 109(42), 17117-17122. https://doi.org/10.1073/pnas.1202079109

Chiniquy, D, Sharma, V, Schultink, A, Baidoo, EE, Rautengarten, C, Cheng, K, Carroll, A, Ulvskov, P, Harholt, J, Keasling, JD, Pauly, M, Scheller, HV & Ronald, PC 2012, 'XAX1 from glycosyltransferase family 61 mediates xylosyltransfer to rice xylan', Proceedings of the National Academy of Sciences of the United States of America, vol. 109, no. 42, pp. 17117-17122. https://doi.org/10.1073/pnas.1202079109

@article{36d7fc3397f34de6bc1466bbfdebce36,

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 residueswith 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 1H-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.",

author = "Dawn Chiniquy and Vaishali Sharma and Alex Schultink and Baidoo, {Edward E.} and Carsten Rautengarten and Kun Cheng and Andrew Carroll and Peter Ulvskov and Jesper Harholt and Keasling, {Jay D.} and Markus Pauly and Scheller, {Henrik V.} and Ronald, {Pamela C.}",

year = "2012",

month = oct,

day = "16",

doi = "10.1073/pnas.1202079109",

language = "English",

volume = "109",

pages = "17117--17122",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "The National Academy of Sciences of the United States of America",

number = "42",

}

TY - JOUR

T1 - XAX1 from glycosyltransferase family 61 mediates xylosyltransfer to rice xylan

AU - Chiniquy, Dawn

AU - Sharma, Vaishali

AU - Schultink, Alex

AU - Baidoo, Edward E.

AU - Rautengarten, Carsten

AU - Cheng, Kun

AU - Carroll, Andrew

AU - Ulvskov, Peter

AU - Harholt, Jesper

AU - Keasling, Jay D.

AU - Pauly, Markus

AU - Scheller, Henrik V.

AU - Ronald, Pamela C.

PY - 2012/10/16

Y1 - 2012/10/16

N2 - 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 residueswith 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 1H-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.

AB - 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 residueswith 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 1H-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.

U2 - 10.1073/pnas.1202079109

DO - 10.1073/pnas.1202079109

M3 - Journal article

SN - 0027-8424

VL - 109

SP - 17117

EP - 17122

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 42

ER -

XAX1 from glycosyltransferase family 61 mediates xylosyltransfer to rice xylan

Abstract

Access to Document

Fingerprint

Cite this