Literature DB >> 18974962

Bioluminescent assay for sphingolipid ceramide N-deacylase using Vibrio harveyi dark mutant M-17.

Ki Woong Cho1.   

Abstract

A new bioluminescent assay method for the activity of sphingolipid ceramide N-deacylase (SCDase: EC 3.5.1.69) as well as ceramidase (CDase: EC 3.5.1.23) was developed using bioluminescent marine bacteria. Enzymatically synthesized ceramide (N-myristoyl sphigosine, C14:0-18:l) and commercial SCDase were used in this demonstration, and myristic (tetradecanoic, C14:0) acid produced by the SCDase hydrolysis was quantified using Vibrio harveyi M-17, a dark mutant of V. harveyi. The in vivo light intensity of M-17 was stimulated up to thousands fold in the presence of myristic acid, was used for this assay. SCDase activity with as little as 10 microU and 5 nM of myristic acid production were detected in less than one min. The assay worked well for the determination of Km and chromatographic fraction assay.

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Year:  2008        PMID： 18974962     DOI： 10.1007/s12275-008-0114-2

Source DB:  PubMed          Journal:  J Microbiol        ISSN： 1225-8873            Impact factor:   3.422


  14 in total

1.  Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower.

Authors:  S Mitsutake; K Kita; T Nakagawa; M Ito
Journal:  J Biochem       Date:  1998-05       Impact factor: 3.387

2.  Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa.

Authors:  N Okino; M Tani; S Imayama; M Ito
Journal:  J Biol Chem       Date:  1998-06-05       Impact factor: 5.157

3.  A stable, inexpensive, solid-state photomultiplier photometer.

Authors:  G W Mitchell; J W Hastings
Journal:  Anal Biochem       Date:  1971-01       Impact factor: 3.365

4.  Chemistry and metabolism of sphingolipids. On the biosynthesis of phytosphingosine by yeast.

Authors:  S R Thorpe; C C Sweeley
Journal:  Biochemistry       Date:  1967-03       Impact factor: 3.162

5.  Activation of bacterial ceramidase by anionic glycerophospholipids: possible involvement in ceramide hydrolysis on atopic skin by Pseudomonas ceramidase.

Authors:  Katsuhiro Kita; Noriyuki Sueyoshi; Nozomu Okino; Masanori Inagaki; Hideharu Ishida; Makoto Kiso; Shuhei Imayama; Takashi Nakamura; Makoto Ito
Journal:  Biochem J       Date:  2002-03-15       Impact factor: 3.857

6.  The separation and direct detection of ceramides and sphingoid bases by normal-phase high-performance liquid chromatography and evaporative light-scattering detection.

Authors:  T J McNabb; A E Cremesti; P R Brown; A S Fischl
Journal:  Anal Biochem       Date:  1999-12-15       Impact factor: 3.365

7.  Large-scale purification and characterization of recombinant Pseudomonas ceramidase: regulation by calcium.

Authors:  Bill X Wu; Christopher F Snook; Motohiro Tani; Erika E Büllesbach; Yusuf A Hannun
Journal:  J Lipid Res       Date:  2006-12-12       Impact factor: 5.922

8.  A novel enzyme that cleaves the N-acyl linkage of ceramides in various glycosphingolipids as well as sphingomyelin to produce their lyso forms.

Authors:  M Ito; T Kurita; K Kita
Journal:  J Biol Chem       Date:  1995-10-13       Impact factor: 5.157

Review 9.  Ceramide in apoptosis: an overview and current perspectives.

Authors:  Benjamin J Pettus; Charles E Chalfant; Yusuf A Hannun
Journal:  Biochim Biophys Acta       Date:  2002-12-30

10.  Differential regulation of sphingomyelinase and ceramidase activities by growth factors and cytokines. Implications for cellular proliferation and differentiation.

Authors:  E Coroneos; M Martinez; S McKenna; M Kester
Journal:  J Biol Chem       Date:  1995-10-06       Impact factor: 5.157
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