A simple and rapid thin-layer chromatography method to follow the kinetics of product formation by 7,8-dihydro-6-hydroxymethylpterin pyrophosphokinase and 7,8-dihydropteroate synthetase
✍ Scribed by L. Bock; W. Butte; M. Richter; J.K. Seydel
- Publisher
- Elsevier Science
- Year
- 1978
- Tongue
- English
- Weight
- 754 KB
- Volume
- 86
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A rapid, simple, and inexpensive thin-layer chromatography method was developed to follow and to quantify enzymatic reactions of the title enzymes without need of a microbiological test system. The method was used to measure the kinetics of product formation by the folate-synthesizing enzymes 7,8-dihydro-6hydroxymethylpterin pyrophosphokinase (I) and 7,8-dihydropteroate synthetase (II) ofEscherichia coli. Thus test conditions could be optimized for the reaction of H,PtCH,OH to H$tCH,OPP by I and of H$tCH,OH to H,-pteroic acid as well as its derivatives by I plus II. Furthermore this tic system allows direct measurements of I and/or II activity in fractions obtained by column chromatography (gel filtration and ion exchange, respectively). ' This work is taken in part from the Ph.D. thesis of W. Butte.