Understanding High-Salt and Cold Adaptation of a Polyextremophilic Enzyme

• Published in: Microorganisms
• Main Authors: Ram Karan, Sam Mathew, Reyhan Muhammad, Didier B. Bautista, Malvina Vogler, Jorg Eppinger, Romina Oliva, Luigi Cavallo, Stefan T. Arold, Magnus Rueping
• Format: Text
• Language: English
• Published: Multidisciplinary Digital Publishing Institute 2020
• Subjects: extremophiles; halophiles; psychrophiles; polyextremophiles; extremozymes; X-ray crystallography; molecular dynamics simulations; Antarc*; Antarctica
• Online Access: https://doi.org/10.3390/microorganisms8101594

The haloarchaeon Halorubrum lacusprofundi is among the few polyextremophilic organisms capable of surviving in one of the most extreme aquatic environments on Earth, the Deep Lake of Antarctica (−18 °C to +11.5 °C and 21–28%, w/v salt content). Hence, H. lacusprofundi has been proposed as a model for biotechnology and astrobiology to investigate potential life beyond Earth. To understand the mechanisms that allow proteins to adapt to both salinity and cold, we structurally (including X-ray crystallography and molecular dynamics simulations) and functionally characterized the β-galactosidase from H. lacusprofundi (hla_bga). Recombinant hla_bga (produced in Haloferax volcanii) revealed exceptional stability, tolerating up to 4 M NaCl and up to 20% (v/v) of organic solvents. Despite being cold-adapted, hla_bga was also stable up to 60 °C. Structural analysis showed that hla_bga combined increased surface acidity (associated with halophily) with increased structural flexibility, fine-tuned on a residue level, for sustaining activity at low temperatures. The resulting blend enhanced structural flexibility at low temperatures but also limited protein movements at higher temperatures relative to mesophilic homologs. Collectively, these observations help in understanding the molecular basis of a dual psychrophilic and halophilic adaptation and suggest that such enzymes may be intrinsically stable and functional over an exceptionally large temperature range.