Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme
The polyextremophilic β-galactosidase enzyme of the haloarchaeon <i>Halorubrum lacusprofundi</i> 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...
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MDPI AG
2022-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/24/15620 |
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| author | Victoria J. Laye Shahlo Solieva Vincent A. Voelz Shiladitya DasSarma |
| author_facet | Victoria J. Laye Shahlo Solieva Vincent A. Voelz Shiladitya DasSarma |
| author_sort | Victoria J. Laye |
| collection | DOAJ |
| description | The polyextremophilic β-galactosidase enzyme of the haloarchaeon <i>Halorubrum lacusprofundi</i> 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 (<i>k<sub>cat</sub></i>/<i>K</i><sub>m</sub>) improves with increasing temperature and salinity, <i>K</i><sub>m</sub> is reduced with decreasing temperatures and increasing salinity, consistent with improved substrate binding at low temperatures. In contrast, <i>k<sub>cat</sub></i> 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. |
| first_indexed | 2024-03-09T16:19:41Z |
| format | Article |
| id | doaj.art-6a6787b084c149a59565ebb8d7d02a3a |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T16:19:41Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-6a6787b084c149a59565ebb8d7d02a3a2023-11-24T15:24:43ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0123241562010.3390/ijms232415620Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic EnzymeVictoria J. Laye0Shahlo Solieva1Vincent A. Voelz2Shiladitya DasSarma3Institute of Marine and Environmental Technology, University System of Maryland, Baltimore, MD 21202, USADepartment of Chemistry, Temple University, Philadelphia, PA 19122, USADepartment of Chemistry, Temple University, Philadelphia, PA 19122, USAInstitute of Marine and Environmental Technology, University System of Maryland, Baltimore, MD 21202, USAThe polyextremophilic β-galactosidase enzyme of the haloarchaeon <i>Halorubrum lacusprofundi</i> 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 (<i>k<sub>cat</sub></i>/<i>K</i><sub>m</sub>) improves with increasing temperature and salinity, <i>K</i><sub>m</sub> is reduced with decreasing temperatures and increasing salinity, consistent with improved substrate binding at low temperatures. In contrast, <i>k<sub>cat</sub></i> 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.https://www.mdpi.com/1422-0067/23/24/15620halophilepsychrophileenzyme kineticsmolecular dynamics simulationsβ-galactosidase |
| spellingShingle | Victoria J. Laye Shahlo Solieva Vincent A. Voelz Shiladitya DasSarma Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme International Journal of Molecular Sciences halophile psychrophile enzyme kinetics molecular dynamics simulations β-galactosidase |
| title | Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme |
| title_full | Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme |
| title_fullStr | Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme |
| title_full_unstemmed | Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme |
| title_short | Effects of Salinity and Temperature on the Flexibility and Function of a Polyextremophilic Enzyme |
| title_sort | effects of salinity and temperature on the flexibility and function of a polyextremophilic enzyme |
| topic | halophile psychrophile enzyme kinetics molecular dynamics simulations β-galactosidase |
| url | https://www.mdpi.com/1422-0067/23/24/15620 |
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