Figure 3

A. Representative paper chromatographic detection of sugars metabolites. The sugars or intermediate metabolites (0.1–0.5 mM) were incubated with BSA (control), PGDM or enolase mutants overnight at room temperature, proteins removed by centrifugation at 10000 × g and an aliquot of supernatant was spotted. a. d. The chromatogram for enolase mutant S39A stained with silver nitrate reagent, b. c. chromatogram for enolase mutant S39A stained with ammonium molybdate reagent. B. Representative thin layer chromatography of sugar metabolites (3-phosphoglycerate, 3-phosphoglyceraldehdye). About 0.1 mM substrate (0.1 mM of each mixture component) as indicated was incubated overnight with purified S39A enolase or BSA (control) has been shown. The chromatogram was developed using silver nitrate reagent following protocol as described in methods and image has been converted to greyscale.