反应详情

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

1. concentrationA cellobiose (Sigma C7252) stock solution of 10 mM final concentration
2. customwas prepared in 50 mM sodium acetate buffer pH=4.5
3. additionFor the assay 2000 μl substrate-stock (10 mM)+100 μL diluted enzyme sample
4. additionwere mixed and incubated at 40° C
5. customwere stopped at t=0, 10, 20 and 30 minutes
6. customequipped with an ED 40 pulsed amperometric detector