Identification and Characterization of Glycosyltransferases Involved in the Biosynthesis of Neodiosmin

Abstract

Glycosylation plays a very important role in plant secondary metabolic modifications. Neodiosmin, identified as diosmetin-7-O-neohesperidoside, not only acts to mitigate bitterness and enhance the flavor of food but also serves as a pivotal metabolite that reinforces plant immunity. Investigating its biosynthetic pathway in plants is crucial for optimizing fruit quality and fortifying plant immune responses. In this study, through analysis of transcriptomic data from Astilbe chinensis, we identified two novel uridine diphosphate (UDP)-glycosyltransferases (UGTs): Ach14791 (AcUGT73C18), responsible for flavonoid 7-Oglycosylation and Ach15849 (AcUGT79B37), involved in flavonoid-7-O-glucoside-2″-O-rhamnosylation. By delving into enzymatic properties and catalytic promiscuity, we developed a biosynthesis route of neodiosmin by establishing a one-pot enzyme-catalyzed cascade reaction. Simultaneously, lonicerin and rhoifolin were also successfully synthesized using the same one-pot dual-enzyme catalytic reaction. Taken together, our findings not only identified two novel UGTs involved in neodiosmin biosynthesis but also provided important biocatalytic components for the microorganism-based biosynthesis of flavonoid-7-O-disaccharide compounds.

Multicopper Oxidase from Lactobacillus hilgardii: Mechanism of Degradation of Tyramine and Phenylethylamine in Fermented Food

“…This resulted in the supernatant containing LHMCO 1614 , which was obtained through further centrifugation at 12,000 g for 30 min at 4 °C. Afterward, the supernatant was introduced into a pre-equilibrated His Trap HP (1 mL) column with a buffer consisting of 50 mM Tris-HCl (pH 7.4) . The extraction of LHMCO 1614 was facilitated by the utilization of an elution buffer composed of 20 mM Tris-HCl, 500 mM imidazole, and 500 mM NaCl at a pH 7.0.…”