|
KMID : 0880220080460050585
|
|
Journal of Microbiology
2008 Volume.46 No. 5 p.585 ~ p.589
|
|
|
Bioluminescent Assay for Sphingolipid Ceramide N-Deacylase Using Vibrio harveyi Dark Mutant M-17
|
|
Cho Ki-Woong
|
|
|
Abstract
|
|
|
A new bioluminescent assay method for the activity of sphingolipid ceramide N-deacylase (SCDase: EC3.5.1.69) as well as ceramidase (CDase: EC 3.5.1.23) was developed using bioluminescent marine bacteria. Enzymatically synthesized ceramide (N-myristoyl sphigosine, C14:0-18:1) 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 μU 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.
|
|
|
KEYWORD
|
|
|
bioluminescent assay, Vibrio harveyi M-17, SCDase, ceramide, myristic acid
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
  
|