A standardized approach for accurate quantification of murein hydrolase activity in high-throughput assays.
Current spectrophotometers measure murein hydrolase activity simultaneously under many conditions and in small intervals. A correct interpretation of these large data sets requires clear and standardized criteria. Furthermore, there is a need for a uniform unit definition to express enzymatic activity, because application of variable definitions seriously hampered comparison between different studies. The method presented here is based on maximizing R(2)-values of incremental data sets. Combined with an appropriate unit definition, it provides a statistically sound background and warrants reproducible and reliable results. Activity calculations are further simplified by an online available Excel spreadsheet. This method is especially suited for experiments where individual curves differ extensively from each other (e.g. low versus high activity conditions) and can be expanded to other similar high-throughput bioassays.
Briers
Yves
Y
Division of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, B-3001 Leuven, Belgium.
Lavigne
Rob
R
Volckaert
Guido
G
Hertveldt
Kirsten
K
D000818:Animals; D009113:Muramidase / Q000032:analysis; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000032:analysis* / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000378:metabolism; D013053:Spectrophotometry / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data
D009113:Muramidase; D009238:N-Acetylmuramoyl-L-alanine Amidase