Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme

Victoria J. Laye, Shahlo Solieva, Vincent Voelz, Shiladitya DasSarma

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

The polyextremophilic β-galactosidase enzyme of the haloarchaeon Halorubrum lacusprofundi functions in extremely cold and hypersaline conditions. To better understand the basis of polyextremophilic activity, the enzyme was studied using steady-state kinetics and molecular dynamics at temperatures ranging from 10 °C to 50 °C and salt concentrations from 1 M to 4 M KCl. Kinetic analysis showed that while catalytic efficiency (kcat/Km) improves with increasing temperature and salinity, Km is reduced with decreasing temperatures and increasing salinity, consistent with improved substrate binding at low temperatures. In contrast, kcat was similar from 2–4 M KCl across the temperature range, with the calculated enthalpic and entropic components indicating a threshold of 2 M KCl to lower the activation barrier for catalysis. With molecular dynamics simulations, the increase in per-residue root-mean-square fluctuation (RMSF) was observed with higher temperature and salinity, with trends like those seen with the catalytic efficiency, consistent with the enzyme’s function being related to its flexibility. Domain A had the smallest change in flexibility across the conditions tested, suggesting the adaptation to extreme conditions occurs via regions distant to the active site and surface accessible residues. Increased flexibility was most apparent in the distal active sites, indicating their importance in conferring salinity and temperature-dependent effects.

Original languageEnglish
Article number15620
JournalInternational Journal of Molecular Sciences
Volume23
Issue number24
DOIs
StatePublished - Dec 2022

Keywords

  • enzyme kinetics
  • halophile
  • molecular dynamics simulations
  • psychrophile
  • β-galactosidase

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