Preparation of riboflavin specifically labeled in the 5′-hydroxymethyl terminus using a vitamin B2-aldehyde-forming enzyme from Schizophyllum commune
✍ Scribed by Tea N. Kekelidze; Dale E. Edmondson; Donald B. McCormick
- Book ID
- 102372142
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- French
- Weight
- 339 KB
- Volume
- 36
- Category
- Article
- ISSN
- 0022-2135
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
A method is described for synthesis of riboflavin selectively labeled in the hydrogens at the 5′‐hydroxymethyl position. In this method, a vitamin B~2~‐aldehyde‐forming enzyme from Schizophyllum commune is used to specifically and completely oxidize the 5′‐hydroxymethyl of riboflavin to the 5′‐aldehyde. This reaction is monitored spectrophotometrically by the reduction of 2,6‐dichlorophenolindophenol at 600 nm. Appearance of aldehyde product was directly quantitated by reverse‐phase high‐performance liquid chromatography. Product is extracted from the incubation mixture by phenol after saturation with (NH~4~)~2~SO~4~ and then further purified by benzyl alcohol extraction. The 5′‐aldehyde is reduced with appropriately labeled sodium borohydride to yield the vitamin specifically labeled in the 5′‐hydroxymethyl group.