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- Title
Cloning and Expression of Gene Encoding a Novel Endoglycoceramidase of Rhodococcus sp. Strain C91.
- Authors
Sakaguchi, Keishi; Okino, Nozomu; Sueyoshi, Noriyuki; Izu, Hiroyuki; Ito, Makoto
- Abstract
Endoglycoceramidase (EGCase) is an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides of various glycosphingolipids. We previously reported that the Asn-Glu-Pro (NEP) sequence is part of the active site of EGCase of Rhodococcus sp. strain M-777. This paper describes the molecular cloning of a new EGCase gene utilising the NEP sequence front the genomic library of Rhodococcus sp. strain C9, which was clearly distinguishable from M-777 by 16S rDNA analysis. C9 EGCase possessed an open reading frame of 1,446 bp encoding 482 amino acids, and showed 78% and 76% identity to M-777 EGCase II at the nucleotide and amino acid levels, respectively. Interestingly, C9 EGCase showed the different specificity to the M-777 enzyme: it hydrolyzed b-series gangliotetraosylceramides more slowly than the M-777 enzyme, whereas both enzymes hydrolyzed a-series gangliosides and neutral glycosphingolipids to the same extent
- Subjects
ENDOGLYCOCERAMIDASE; AMINO acids; RHODOCOCCUS; ENZYMES; ENZYMOLOGY
- Publication
Journal of Biochemistry, 2000, Vol 128, Issue 1, p145
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/oxfordjournals.jbchem.a022725