ArticlePloS one2016
Statistical Evaluation of HTS Assays for Enzymatic Hydrolysis of β-Keto Esters.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 12 citations in OpenAlex.
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- Developing a fluorometric urease activity microplate assay suitable for automated microbioreactor experiments.Frontiers in bioengineering and biotechnology · 2022Article
- Design of Experiments As a Tool for Optimization in Recombinant Protein Biotechnology: From Constructs to Crystals.Molecular biotechnology · 2019Review
- β-Phenylalanine Ester Synthesis from Stable β-Keto Ester Substrate Using Engineered ω-Transaminases.Molecules (Basel, Switzerland) · 2018Article
- Riboswitching with ciprofloxacin-development and characterization of a novel RNA regulator.Nucleic acids research · 2018Article
- StreAM-[Formula: see text]: algorithms for analyzing coarse grained RNA dynamics based on Markov models of connectivity-graphs.Algorithms for molecular biology : AMB · 2017Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
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.
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