Development of Fluorogenic Probes to Detect Glycan-Degrading Enzymes in Bacteria

Area of Science:

• Microbiology
• Biochemistry
• Glycobiology

Background:

• Bacterial cell-envelope glycans are vital for fitness and survival.
• Their synthesis and degradation involve numerous glycosyltransferases and glycosidases.
• Limited methods exist to profile bacterial glycosidase activity, leaving many functions unknown.

Purpose of the Study:

• To develop and validate novel fluorogenic probes for detecting bacterial glycosidase activity.
• To expand the toolkit for studying bacterial glycan metabolism.
• To gain insights into the diversity and function of unexplored bacterial glycan-processing enzymes.

Main Methods:

• Development of fluorogenic probes based on bacterial specific sugars: N-acetyl-d-fucosamine and 2,4-diacetamido-2,4,6-trideoxy-d-galactose.
• Comparison of probe activation with a d-glucose-based probe in bacterial and mammalian cells.
• Analysis of species-selective probe utilization and relative glycosidase activity.

Main Results:

• Successful development of novel fluorogenic probes for bacterial glycosidases.
• Demonstrated species-selective utilization of the probes.
• Observed differences in the magnitude of glycosidase activity across species.
• Provided insights into unexplored glycan-processing enzymes.

Conclusions:

• Novel fluorogenic probes enhance the detection of glycan-degrading enzymes in bacteria.
• These probes offer a new approach to understanding bacterial glycan metabolism and its diversity.
• The findings contribute to a broader understanding of bacterial cell-envelope maintenance and function.