Cloning and expression of gene encoding a novel endoglycoceramidase of Rhodococcus sp. strain C9

Keishi Sakaguchi, Nozomu Okino, Noriyuki Sueyoshi, Hiroyuki Izu, Makoto Ito

Research output: Contribution to journalArticle

9 Citations (Scopus)

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 utilizing the NEP sequence from 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.

Original languageEnglish
Pages (from-to)145-152
Number of pages8
JournalJournal of biochemistry
Volume128
Issue number1
DOIs
Publication statusPublished - Jan 1 2000

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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