Overexpression in a non-native halophilic host and biotechnological potential of NAD +-dependent glutamate dehydrogenase from Halobacterium salinarum strain NRC-36014

Nayla Munawar, Paul C. Engel

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

Enzymes produced by halophilic archaea are generally heat resistant and organic solvent tolerant, and accordingly important for biocatalytic applications in 'green chemistry', frequently requiring a low-water environment. NAD +-dependent glutamate dehydrogenase from an extremely halophilic archaeon Halobacterium salinarum strain NRC-36014 was selected to explore the biotechnological potential of this enzyme and genetically engineered derivatives. Over-expression in a halophilic host Haloferax volcanii provided a soluble, active recombinant enzyme, not achievable in mesophilic Escherichia coli, and an efficient purification procedure was developed. pH and salt dependence, thermostability, organic solvent stability and kinetic parameters were explored. The enzyme is active up to 90 °C and fully stable up to 70 °C. It shows good tolerance of various miscible organic solvents. High concentrations of salt may be substituted with 30 % DMSO or betaine with good stability and activity. The robustness of this enzyme under a wide range of conditions offers a promising scaffold for protein engineering.

Original languageEnglish
Pages (from-to)463-476
Number of pages14
JournalExtremophiles
Volume16
Issue number3
DOIs
Publication statusPublished - May 2012
Externally publishedYes

Keywords

  • Glutamate dehydrogenase
  • Halobacterium
  • Halophilic enzyme
  • Halophilic expression host
  • Organic solvents
  • Salt substitution
  • Thermostability

ASJC Scopus subject areas

  • Microbiology
  • Molecular Medicine

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