Direct measurement of ascorbic acid biosynthesis in Arabidopsis cell suspension culture using capillary electrophoresis.
We describe procedures to directly measure the biosynthesis of vitamin C (L-ascorbic acid, L-AA) in crude extracts of an Arabidopsis thaliana cell suspension culture by capillary electrophoresis. Optimal conditions have been established for the quantitation of L-AA formed by the oxidation of three different substrates: L-galactose, L-galactono-1,4-lactone, and L-gulono-1,4-lactone. We also demonstrate that L-galactono-1,4-lactone dehydrogenase activity does not require exogenous cofactor. The minimal sample handling requirements, the high selectivity, and short analysis times represent significant advantages over existing protocols.
Davey
M W
MW
Departement Plantengenetica, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Universiteit Gent, Belgium.
Persiau
G
G
Bauw
G
G
Van Montagu
M
M
D017360:Arabidopsis / Q000737:chemistry*; D001205:Ascorbic Acid / Q000032:analysis* / Q000096:biosynthesis; D002478:Cells, Cultured; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D010088:Oxidoreductases / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D013379:Substrate Specificity
D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C093910:galactonolactone dehydrogenase; D001205:Ascorbic Acid