Sodium Hydroxide → D-(+)-Cellobiose (CNP0226604.15)
Reaction equation
[OH-:1].[Na+].[O:3]=[CH:4][C@@H:5]([C@H:7]([C@@H:9]([C@@H:11]([CH2:13][OH:14])[OH:12])[OH:10])[OH:8])[OH:6]>>[CH2:13]([OH:14])[C@H:11]1[O:12][C@@H:4]([O:3][C@H:9]2[C@H:7]([OH:8])[C@@H:5]([OH:6])[C@H:4]([OH:3])[O:1][C@@H:11]2[CH2:13][OH:14])[C@H:5]([OH:6])[C@@H:7]([OH:8])[C@@H:9]1[OH:10]Participants
3
Procedure
A cellobiose (Sigma C7252) stock solution of 10 mM final concentration was prepared in 50 mM sodium acetate buffer pH=4.5. For the assay 2000 μl substrate-stock (10 mM)+100 μL diluted enzyme sample were mixed and incubated at 40° C. Reactions were stopped at t=0, 10, 20 and 30 minutes by mixing 100 μl incubate+100 μl 50 mM sodiumhydroxide. Samples were subjected to ultrafiltration and analyzed using High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection (HPAEC-PAD), performed on a Dionex DX-500 equipped with an ED 40 pulsed amperometric detector. Activity is expressed in μmol glucose released/ml/min
Workup
- concentrationA cellobiose (Sigma C7252) stock solution of 10 mM final concentration