| RRC ID |
48010
|
| Author |
Katafuchi Y, Li Q, Tanaka Y, Shinozuka S, Kawamitsu Y, Izumi M, Ekino K, Mizuki K, Takegawa K, Shibata N, Goto M, Nomura Y, Ohta K, Oka T.
|
| Title |
GfsA is a β1,5-galactofuranosyltransferase involved in the biosynthesis of the galactofuran side chain of fungal-type galactomannan in Aspergillus fumigatus.
|
| Journal |
Glycobiology
|
| Abstract |
Previously, we reported that GfsA is a novel galactofuranosyltransferase involved in the biosynthesis of O-glycan, the proper maintenance of fungal morphology, the formation of conidia and anti-fungal resistance in Aspergillus nidulans and A. fumigatus (Komachi Y et al., 2013. GfsA encodes a novel galactofuranosyltransferase involved in biosynthesis of galactofuranose antigen of O-glycan in Aspergillus nidulans and Aspergillus fumigatus. Mol. Microbiol. 90:1054-1073). In the present paper, to gain an in depth-understanding of the enzymatic functions of GfsA in A. fumigatus (AfGfsA), we established an in vitro assay to measure galactofuranosyltransferase activity using purified AfGfsA, UDP-α-d-galactofuranose as a sugar donor, and p-nitrophenyl-β-d-galactofuranoside as an acceptor substrate. LC/MS, 1H-NMR and methylation analyses of the enzymatic products of AfGfsA revealed that this protein has the ability to transfer galactofuranose to the C-5 position of the β-galactofuranose residue via a β-linkage. AfGfsA requires a divalent cation of manganese for maximal activity and consumes UDP-α-d-galactofuranose as a sugar donor. Its optimal pH range is 6.5-7.5 and its optimal temperature range is 20-30°C. 1H-NMR, 13C-NMR and methylation analyses of fungal-type galactomannan extracted from the ∆AfgfsA strain revealed that AfGfsA is responsible for the biosynthesis of β1,5-galactofuranose in the galactofuran side chain of fungal-type galactomannan. Based on these results, we conclude that AfGfsA acts as a UDP-α-d-galactofuranose: β-d-galactofuranoside β1,5-galactofuranosyltransferase in the biosynthetic pathway of galactomannans.
|
| Volume |
27(6)
|
| Pages |
568-581
|
| Published |
2017-6-1
|
| DOI |
10.1093/glycob/cwx028
|
| PII |
3091656
|
| PMID |
28369326
|
| MeSH |
Aspergillus fumigatus / enzymology*
Fungal Polysaccharides / chemistry
Fungal Polysaccharides / metabolism*
Fungal Proteins / chemistry
Fungal Proteins / genetics
Fungal Proteins / metabolism*
Furans / chemistry
Furans / metabolism
Galactose / analogs & derivatives
Galactosyltransferases / chemistry
Galactosyltransferases / genetics
Galactosyltransferases / metabolism*
Manganese / chemistry
Mannans / chemistry
Mannans / metabolism
|
| IF |
4.06
|
| Times Cited |
14
|
| Resource |
| Prokaryotes E. coli |
JW2021-AM |
