Development and application of a suite of polysaccharide-degrading enzymes for analyzing plant cell walls

被引:252
作者
Bauer, Stefan
Vasu, Prasanna
Persson, Staffan
Mort, Andrew J.
Somerville, Chris R. [1 ]
机构
[1] Carnegie Inst, Stanford, CA 94305 USA
[2] Oklahoma State Univ, Dept Biochem & Mol Biol, Stillwater, OK 74078 USA
关键词
Arabidopsis; fingerprinting; hydrolase; mutant; xylan;
D O I
10.1073/pnas.0604632103
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
To facilitate analysis of plant cell wall polysaccharide structure and composition, we cloned 74 genes encoding polysaccharicle-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 polysaccharicles 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.
引用
收藏
页码:11417 / 11422
页数:6
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