Bicarbonate Ion → Biotin
Reaction equation
[C:1](=[O:4])(O)[O-:2].[CH2:5]1[S:9][C@@H:8]([CH2:10][CH2:11][CH2:12][CH2:13]C(NCCCCCC(ON2C(=O)C(S([O-])(=O)=O)CC2=O)=O)=O)[C@H:7]2[NH:36][C:37]([NH:39][C@@H:6]12)=[O:38].[Na+]>O>[OH:2][C:1]([CH2:13][CH2:12][CH2:11][CH2:10][C@H:8]1[C@@H:7]2[C@@H:6]([NH:39][C:37]([NH:36]2)=[O:38])[CH2:5][S:9]1)=[O:4]Participants
4
Reactant 2
Product 1
Reagents, catalysts & solvents 1
Procedure
About 2.0 mg of DBP is buffer exchanged with 20 mM bicarbonate at pH 9.6 three times to give a final volume of 1.0 mL. Two aliquots of sulfo-NHS-LC-biotin (Pierce) are dissolved in water to give a 1 mg/mL solution just prior to addition. The first aliquot (10 uL) is added and the reaction is maintained at room temperature for 5 minutes. The second aliquot (10 uL) is added and the reaction proceeds for an additional 4 minutes. The unreacted biotin is removed by size exclusion chromatography using SEPHADEX G-25 and 100 mM PBS, pH 7.4 as the mobile phase.