Development and application of a suite of polysaccharide-degrading enzymes for analyzing plant cell walls.
To facilitate analysis of plant cell wall polysaccharide structure and composition, we cloned 74 genes encoding polysaccharide-degrading enzymes from Aspergillus nidulans, Aspergillus fumigatus, and Neurospora crassa and expressed the genes as secreted proteins with C-terminal Myc and 6x His tags. Most of the recombinant enzymes were active in enzyme assays, and optima for pH and temperature were established. A subset of the enzymes was used to fragment polysaccharides from the irregular xylem 9 (irx9) mutant of Arabidopsis. The analysis revealed a decrease in the abundance of xylan in the mutant, indicating that the IRX9 gene, which encodes a putative family 43 glycosyltransferase, is required for xylan synthesis.
Bauer
Stefan
S
Carnegie Institution, Stanford, CA 94305, USA.
Vasu
Prasanna
P
Persson
Staffan
S
Mort
Andrew J
AJ
Somerville
Chris R
CR
D017360:Arabidopsis / Q000378:metabolism; D001232:Aspergillus fumigatus / Q000378:metabolism; D001233:Aspergillus nidulans / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005821:Genetic Techniques; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry*; D013696:Temperature; D014990:Xylans / Q000737:chemistry
D010940:Plant Proteins; D011134:Polysaccharides; D014990:Xylans