Multiply mutated Gaussia luciferases provide prolonged and intense bioluminescence

Biochem Biophys Res Commun. 2009 Nov 27;389(4):563-8. doi: 10.1016/j.bbrc.2009.09.006.


Gaussia luciferase (GLuc) from the copepod Gaussia princeps is both the smallest and brightest known luciferase. GLuc catalyzes the oxidation of coelenterazine to produce an intense blue light but with a very short emission half-life. We report mutated GLucs with much longer luminescence half-lives that retain the same initial intensity as the wild-type enzyme. The GLuc variants were produced using cell-free protein synthesis to provide high yields and rapid production of fully active product as well as simple non-natural amino acid substitution. By incorporating homopropargylglycine and attaching PEG using azide-alkyne click reactions, we also show that the four methionines in GLuc are surface accessible. The mutants provide a significantly improved reporter protein for both in vivo and in vitro studies, and the successful non-natural amino acid incorporation and PEG attachment indicate the feasibility of producing useful bioconjugates using click attachment reactions.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Copepoda / enzymology*
  • Luciferases / chemistry*
  • Luciferases / genetics
  • Luciferases / isolation & purification
  • Luminescence*
  • Molecular Sequence Data
  • Mutant Proteins / chemistry*
  • Mutant Proteins / genetics
  • Mutant Proteins / isolation & purification
  • Mutation


  • Mutant Proteins
  • Luciferases