Efficient One-Pot Multienzyme Synthesis of β-Glycosides

Area of Science:

• Biocatalysis
• Synthetic Biology
• Enzyme Engineering

Background:

• Phenolic glycosides are vital in medicine and food science.
• Traditional chemical glycosylation methods are complex and inefficient.

Purpose of the Study:

• To develop an efficient one-pot multienzyme cascade system for synthesizing phenolic glycoside derivatives.
• To engineer glycosyltransferases via directed evolution for improved synthesis.
• To establish an environmentally friendly biosynthesis route for glycosides.

Main Methods:

• Constructed a three-module one-pot multienzyme cascade system (UDP-sugar synthesis, UTP regeneration, glycoside synthesis).
• Employed directed evolution for glycosyltransferase optimization.
• Validated the system for both β-glucuronide and β-glucoside synthesis.

Main Results:

• Achieved high yields of β-glucuronide (5.95 g/L from p-nitrophenol) and β-glucoside (8.13 g/L from p-nitrophenol).
• Demonstrated large-scale product formation using catalytic amounts of UTP.
• Confirmed the system's versatility for various phenolic glycoside compounds.

Conclusions:

• The developed multienzyme system provides an efficient and environmentally friendly method for phenolic glycoside production.
• This strategy enables scalable biosynthesis of valuable glycoside compounds.
• Directed evolution and system optimization are key to successful biocatalytic synthesis.