Genetic Engineering Breakthrough in Rhamnosyl and Allosyl Flavonoids Production from Escherichia coli

Scientists from Sun Moon University have successfully engineered Escherichia coli to produce rhamnosyl and allosyl flavonoids, a crucial step in the bioengineering of glycosylated compounds. The researchers used a genetic engineering approach to overexpress TDP-L-rhamnose and TDP-6-deoxy-D-allose biosynthetic gene clusters, and flavonoids were glycosylated by the overexpression of the glycosyltransferase gene from Arabidopsis thaliana. The products were isolated, analyzed, and confirmed by HPLC, LC/MS, and ESI-MS/MS analyses.

Key Takeaways:

• The study reports a genetic engineering approach for the production of rhamnosyl and allosyl flavonoids from Escherichia coli.
• The researchers engineered Escherichia coli by overexpressing TDP-L-rhamnose and TDP-6-deoxy-D-allose biosynthetic gene clusters.
• Flavonoids (quercetin and kaempferol) were exogenously fed to the host in a biotransformation system for glycosylation.
• Several conditions were optimized to increase the production level of glycosylated compounds.
• The researchers successfully isolated approximately 24 mg/L 3-O-rhamnosyl quercetin, 12.9 mg/L 3-O-rhamnosyl kaempferol, and 3-O-allosyl quercetin.
• The study reveals a method useful for the biosynthesis of rhamnosyl and allosyl flavonoids and for the glycosylation of related compounds.

Statistics:

• Approximately 24 mg/L 3-O-rhamnosyl quercetin was isolated upon feeding of 0.2 mM of the respective flavonoids.
• Approximately 12.9 mg/L 3-O-rhamnosyl kaempferol was isolated upon feeding of 0.2 mM of the respective flavonoids.
• 3-O-allosyl quercetin was isolated using the optimized conditions.

Sources:

• Simkhada, D., et al. "Genetic engineering approach for the production of rhamnosyl and allosyl flavonoids from Escherichia coli." Biotechnology & Bioengineering 107.1 (2010): 154-62.