A novel polysaccharide hydrolase cDNA (celD) from Neocallimastix patriciarum encoding three multi-functional catalytic domains with high endoglucanase, cellobiohydrolase and xylanase activities.
A plant polysaccharide hydrolase cDNA, designated celD, was isolated from a cDNA library of the rumen fungus Neocallimastix patriciarum. The enzyme encoded by celD had endoglucanase, cellobiohydrolase and xylanase activities. Deletion analysis revealed that celD cDNA can be truncated to code for three catalytically active domains. Each domain had the same substrate specificity as the enzyme produced by the untruncated celD and also possessed cellulose-binding capacity. Substrate competition studies showed that carboxymethylcellulose and xylan appear to compete with methylumbelliferyl cellobioside for the same active site within each domain. Expression of celD transcript in the rumen fungus was constitutive and was not affected by the presence of cellulose in the culture medium.
Xue
G P
GP
CSIRO Division of Tropical Crops and Pastures, St Lucia, Qld, Australia.
Gobius
K S
KS
Orpin
C G
CG
D000818:Animals; D002266:Carboxymethylcellulose Sodium / Q000378:metabolism; D002475:Cellobiose / Q000031:analogs & derivatives / Q000378:metabolism; D002480:Cellulase / Q000235:genetics; D043366:Cellulose 1,4-beta-Cellobiosidase; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D005658:Fungi / Q000201:enzymology / Q000235:genetics*; D005800:Genes, Fungal / Q000235:genetics*; D006026:Glycoside Hydrolases / Q000235:genetics*; D011134:Polysaccharides / Q000378:metabolism*; D011487:Protein Conformation; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins; D015183:Restriction Mapping; D012417:Rumen / Q000382:microbiology; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D043325:Xylan Endo-1,3-beta-Xylosidase; D014990:Xylans / Q000378:metabolism
D011134:Polysaccharides; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014990:Xylans; D002475:Cellobiose; C068475:4-methylumbelliferylcellobioside; D006026:Glycoside Hydrolases; D043325:Xylan Endo-1,3-beta-Xylosidase; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase; D002266:Carboxymethylcellulose Sodium