A new cold-active transglutaminase: Discovery, computational insights, and recombinant expression

Area of Science:

• Biochemistry
• Enzymology
• Biotechnology

Background:

• Transglutaminases (TGases) are crucial enzymes with broad applications.
• Cold-active TGases are vital for low-temperature bioprocessing but remain underexplored.
• Antarctic krill is a potential source for novel psychrophilic enzymes.

Purpose of the Study:

• To identify and characterize a novel cold-active transglutaminase (akTGase) from Antarctic krill.
• To computationally predict structural features related to cold adaptation.
• To express and validate the enzymatic activity of recombinant akTGase at low temperatures.

Main Methods:

• Transcriptomic mining of Antarctic krill for TGase genes.
• Bioinformatic and molecular dynamics simulations for structural analysis.
• Recombinant expression in E. coli and enzymatic activity assays at different temperatures.

Main Results:

• Identified and computationally characterized akTGase, revealing cold-adaptation structural traits.
• Molecular dynamics simulations indicated enhanced flexibility and stability at 4°C.
• Recombinant akTGase exhibited significantly higher specific activity at 4°C compared to 25°C.

Conclusions:

• akTGase is a novel, cold-active enzyme with potential for low-temperature biocatalysis.
• The study demonstrates an effective strategy for discovering psychrophilic enzymes.
• This research opens avenues for sustainable applications in cold-adapted biotechnology.