KIT | KIT-Bibliothek | Impressum | Datenschutz

Statistical Evaluation of HTS Assays for Enzymatic Hydrolysis of β-Keto Esters

Buß, O. 1; Jager, S.; Dold, S. M. 1; Zimmermann, S. 1; Hamacher, K.; Schmitz, K.; Rudat, J. 1
1 Karlsruher Institut für Technologie (KIT)

Abstract:

β-keto esters are used as precursors for the synthesis of β-amino acids, which are building blocks for some classes of pharmaceuticals. Here we describe the comparison of screening procedures for hydrolases to be used for the hydrolysis of β-keto esters, the first step in the preparation of β-amino acids. Two of the tested high throughput screening (HTS) assays depend on coupled enzymatic reactions which detect the alcohol released during ester hydrolysis by luminescence or absorption. The third assay detects the pH shift due to acid formation using an indicator dye. To choose the most efficient approach for screening, we assessed these assays with different statistical methods—namely, the classical Z’-factor, standardized mean difference (SSMD), the Kolmogorov-Smirnov-test, and t-statistics. This revealed that all three assays are suitable for HTS, the pH assay performing best. Based on our data we discuss the explanatory power of different statistical measures. Finally, we successfully employed the pH assay to identify a very fast hydrolase in an enzyme-substrate screening.


Verlagsausgabe §
DOI: 10.5445/IR/1000051700
Originalveröffentlichung
DOI: 10.1371/journal.pone.0146104
Scopus
Zitationen: 9
Web of Science
Zitationen: 9
Dimensions
Zitationen: 10
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Bio- und Lebensmitteltechnik (BLT)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2016
Sprache Englisch
Identifikator ISSN: 1932-6203
urn:nbn:de:swb:90-517000
KITopen-ID: 1000051700
Erschienen in PLoS one
Verlag Public Library of Science (PLoS)
Band 11
Heft 1
Seiten e0146104
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Nachgewiesen in Web of Science
Scopus
Dimensions
KIT – Die Forschungsuniversität in der Helmholtz-Gemeinschaft
KITopen Landing Page