title: Super RLuc8: A novel engineered Renilla luciferase with a red-shifted spectrum and stable light emission.
abstract: Renilla luciferase is a bioluminescent enzyme which is broadly used as a reporter protein in molecular biosensors. In this study, a novel luciferase with desired light emission wavelength and thermostability is reported. The results indicated that the new luciferase, namely super RLuc8, had a red-shifted spectrum and showed stable light emission. Super RLuc8 showed a 10-fold (p-value=0.0084) increase in the thermostability at 37°C after 20min incubation, in comparison to the native enzyme. The optimum temperature of the mutant increased from 30 to 37°C. Molecular dynamics simulation analysis indicated that the increased thermostability was most probably caused by a better structural compactness and more local rigidity in the regions out of the emitter site.
authors.affiliation: Department of Biology, Nourdanesh Institute of Higher Education, Meymeh, Iran.
authors.forename: Somaieh
authors.initials: S
authors.lastname: Rahnama
authors.affiliation: Department of Genetics, Shahrekord University, Shahrekord, Iran.
authors.forename: Behnaz
authors.initials: B
authors.lastname: Saffar
authors.affiliation: Department of Biology, University of Isfahan, Isfahan, Iran.
authors.forename: Zahra Fanaei
authors.initials: ZF
authors.lastname: Kahrani
authors.affiliation: Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
authors.forename: Mahboobeh
authors.initials: M
authors.lastname: Nazari
authors.affiliation: Department of Biology, University of Isfahan, Isfahan, Iran. Electronic address: Sci_rahman@yahoo.com.
authors.forename: Rahman
authors.initials: R
authors.lastname: Emamzadeh
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001709:Biotechnology; D004795:Enzyme Stability / Q000235:genetics; D007700:Kinetics; D049410:Luciferases, Renilla / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D048790:Renilla / Q000201:enzymology / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D049410:Luciferases, Renilla
keywords: Emission spectrum; Enzyme kinetics; MD simulation; Renilla luciferase; Super RLuc8