Réaction #366023

Équation de réaction

Tris-HCl

desthiobiotin

L-cysteine

DTT

→

biotin

Conditions de réaction

Traitement

1. 1
   AutreThe enzyme reaction mixture without cell-free extracts
2. 2
   AutreThe enzyme reaction mixture in a 300 μl glass spitz tube
3. 3
   AutreThe reaction was started by light irradiation from 10 cm distance with 20 W fluorescent bulb at 30° C
4. 4
   AutreAfter 80 min reaction, reaction

Mode opératoire

The enzyme reaction mixture without cell-free extracts contained 100 μM desthiobiotin. 1000 μM SAM, 200 μM L-cysteine, 50 μM deazariboflavin, 0.6 mg/ml (16 μM) BIOB protein, 10 mM DTT and 0.1 M Tris-HCl/pH 7.5 in total volume of 50 μl. The enzyme reaction mixture in a 300 μl glass spitz tube was brought to anaerobic condition by repeating of weak aspiration and argon pressure under dark. The reaction was started by light irradiation from 10 cm distance with 20 W fluorescent bulb at 30° C. After 80 min reaction, reaction was stopped by heating at 95° C., and produced biotin was determined by the microbiological assay 1 ) using Lactobacillis plantarum. (ATCC8014)

Source