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Nieuwenhuizen, W.F. (author), van Leeuwen, S. (author), Jack, R.W. (author), Egmond, M.R. (author), Götz, F. (author)
Ceramidase (CDase) hydrolyzes the amide bond in ceramides to yield free fatty acid and sphingosine. From a 3-L Pseudomonas aeruginosa PA01 culture, 70 μg of extracellular alkaline, Ca2+-dependent CDase, was purified to homogeneity, the N-terminal sequence was determined, and the CDase gene was cloned. The CDase gene encodes a 670 amino acid...
article 2003
Nieuwenhuizen, W.F. (author), van Leeuwen, S. (author), Götz, F. (author), Egmond, M.R. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)
Ceramidase (CDase) hydrolyses the N-acyl linkage of the sphingolipid ceramide. We synthesized the non-fluorescent ceramide analogue (4E,2S,3R)-2-N-(10-pyrenedecanoyl)-1,3,17-trihydroxy-17- (3,5-dinitrobenzoyl)-4-heptadecene (10) that becomes fluorescent upon hydrolysis of its N-acyl bond. This novel substrate was used to study several kinetic...
article 2002