Expression of a soluble truncated Vargula luciferase in Escherichia coli

Eric A. Hunt, Angeliki Moutsiopoulou, David Broyles, Trajen Head, Emre Dikici, Sylvia Daunert, Sapna K. Deo

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Marine luciferases are regularly employed as useful reporter molecules across a range of various applications. However, attempts to transition expression from their native eukaryotic environment into a more economical prokaryotic, i.e. bacterial, expression system often presents several challenges. Specifically, bacterial protein expression inherently lacks chaperone proteins to aid in the folding process, while Escherichia coli presents a reducing cytoplasmic environment in. These conditions contribute to the inhibition of proper folding of cysteine-rich proteins, leading to incorrect tertiary structure and ultimately inactive and potentially insoluble protein. Vargula luciferase (Vluc) is a cysteine-rich marine luciferase that exhibits glow-type bioluminescence through a reaction between its unique native substrate and molecular oxygen. Because most other commonly used bioluminescent proteins exhibit flash-type emission kinetics, this emission characteristic of Vluc is desirable for high-throughput applications where stability of emission is required for the duration of data collection. A truncated form of Vluc that retains considerable bioluminescence activity (55%) compared to the native full-length protein has been reported in the literature. However, expression and purification of this luciferase from bacterial systems has proven difficult. Herein, we demonstrate the expression and purification of a truncated form of Vluc from E. coli. This truncated Vluc (tVluc) was subsequently characterized in terms of both its biophysical and bioluminescence properties.

Original languageEnglish (US)
Pages (from-to)68-74
Number of pages7
JournalProtein Expression and Purification
Volume132
DOIs
StatePublished - Apr 1 2017

Keywords

  • Bacterial expression
  • Bioluminescence
  • Truncated protein
  • Vargula luciferase

ASJC Scopus subject areas

  • Biotechnology

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