Journal: Journal of Agricultural and Food Chemistry

Article Title: Discovery of Potent Glycosidases Enables Quantification of Smoke-Derived Phenolic Glycosides through Enzymatic Hydrolysis

doi: 10.1021/acs.jafc.4c01247

Figure Lengend Snippet: Initial screening of active glycosidases (GHs) on guaiacol glucoside 1a and guaiacol gentiobioside 1b through genome mining. (A) Sequence similarity network (SSN) of GH1 enzyme family. Only the groups containing the tested sequences are depicted in the figure, while the complete SSN can be found in the Supporting Information . The most active GHs located in representative nodes A in group 1 and B, C in group 4. (B) Utilization of LC–MS analysis for activity screening in wine. This figure demonstrates the application of this method using Cb BglB-1 as a representative example. The semiquantitative activity could be evaluated by comparing total ion counts in MS between samples with the added enzyme and those without it. (C) Semiquantitative heatmap of the degrees of conversion by GH1 enzymes on 1a and 1b in buffer and wine. Candidates such as Cb BglB-1 were mixed with baseline wine which had been fortified with 4.5 mg/L each of compounds 1a and 1b . The reaction was at 37 °C for 4 h duration. The high-activity clusters corresponding to the SSN are also labeled as stars. The most promising candidate Cb BglB-1 is highlighted in red.

Article Snippet: The Interpro IPR001360 collection of GH1 enzyme sequences combined with JGI IMG Integrated Microbial Genomes & Microbiomes database annotated GH1 enzymes was used as the input for EFI-EST analysis of GH1 while Interpro IPR001547 annotated as rutinosidase was used as the input for GH5.

Techniques: Sequencing, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Labeling