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Bioresources and Bioprocessing
Fig. 4 | Bioresources and Bioprocessing

Fig. 4

From: A method for high-throughput screening hydrolase of lignin β-aryl ether linkage from directed evolution by glutathione (GSH) assay

Fig. 4

GSH changes with time between control and test group after optimizing the LigF reaction system. In this part, the total volume of reaction mixture was 5 mL. The test group including 300 μL 20 mM GVG, 200 μL 3 mg/mL GSH, 3800 μL 50 mM Tris–Cl (pH 8.0), 700 μL cell lysate containing the protein of 0.5 mg/mL. While in the control group, the cell lysate was replaced by 700 μL 50 mM Tris–Cl buffer (pH 8.0). The reaction was carried out at 27 °C and 80 rpm. And then the equal volume of ethyl acetate was added to the reaction mixture to stop the enzymatic reaction. The mixture was centrifuged for 2 min at 13000 rpm and 50 μL water phase was taken for the GSH content analysis to describe the LigF activity

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