title: A standardized approach for accurate quantification of murein hydrolase activity in high-throughput assays.
abstract: 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.
authors.affiliation: Division of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, B-3001 Leuven, Belgium.
authors.forename: Yves
authors.initials: Y
authors.lastname: Briers
authors.forename: Rob
authors.initials: R
authors.lastname: Lavigne
authors.forename: Guido
authors.initials: G
authors.lastname: Volckaert
authors.forename: Kirsten
authors.initials: K
authors.lastname: Hertveldt
mesh_terms: 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
chemical_list: D009113:Muramidase; D009238:N-Acetylmuramoyl-L-alanine Amidase