Activity-based protein profiling as a robust method for enzyme identification and screening in extremophilic Archaea

Area of Science:

• Microbiology
• Biochemistry
• Molecular Biology

Background:

• Archaea inhabit extreme environments, posing challenges for in vivo research.
• Existing methodologies limit the thorough investigation of archaeal enzymes.

Purpose of the Study:

• To demonstrate the utility of in vivo activity-based protein profiling (ABPP) for studying archaea.
• To develop ABPP as a tool for functional enzyme screening in archaeal (meta)genomes.

Main Methods:

• In vivo activity-based protein profiling (ABPP) was applied to living archaea.
• A genetically engineered archaeal screening strain was developed for enhanced screening capabilities.

Main Results:

• ABPP successfully detected native and heterogeneously expressed active enzymes in archaea under extreme conditions.
• The developed ABPP approach facilitated functional enzyme screening from (meta)genome samples.

Conclusions:

• In vivo ABPP is a sensitive method for detecting active enzymes in archaea, even in extreme conditions.
• This approach supports basic archaeal research and the discovery of novel enzymes from genomic libraries.