1998年6月5日
Purification and characterization of a novel ceramidase from pseudomonas aeruginosa
Journal of Biological Chemistry
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- ,
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- 巻
- 273
- 号
- 23
- 開始ページ
- 14368
- 終了ページ
- 14373
- 記述言語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1074/jbc.273.23.14368
We report here a novel type of ceramidase of Pseudomonas aeruginosa AN17 isolated from the skin of a patient with atopic dermatitis. The enzyme was purified 83,400-fold with an overall yield of 21.1% from a culture supernatant of strain AN17. After being stained with a silver staining solution, the purified enzyme showed a single protein band, and its molecular mass was estimated to be 70 kDa on SDS-polyacrylamide gel electrophoresis. The enzyme showed quite wide specificity for various ceramides, i.e. it hydrolyzed ceramides containing C12:0-C18:0 fatty acids and 7-nitrobenz-2- oxa-1,3-4-diazole-labeled dodecanoic acid, and not only ceramide containing sphingosine (d18:l) or sphinganine (d18:0) but also phytosphingosine (t18:0) as the long-chain base. However, the enzyme did not hydrolyze galactosylceramide, sulfatide, GM1, or sphingomyelin, and thus was clearly distinguished from a Pseudomonas sphingolipid ceramide N-deacylase (Ito, M., Kurita, T., and Kita, K. (1995) J. Biol. Chem. 270, 24370-24374). This bacterial ceramidase had a pH optimum of 8.0-9.0, an apparent K(m) of 139 μM, and a V(max) of 5.3 μmol/min/mg using N-palmitoylsphingosine as the substrate. The enzyme appears to require Ca2+ for expression of the activity. Interestingly, the 70-kDa protein catalyzed a reversible reaction in which the N-acyl linkage of ceramide was either cleaved or synthesized. Our study demonstrated that ceramidase is widely distributed from bacteria to mammals.
- リンク情報
- ID情報
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- DOI : 10.1074/jbc.273.23.14368
- ISSN : 0021-9258
- PubMed ID : 9603946
- SCOPUS ID : 0032486372