Mutant firefly luciferase enzymes resistant to the inhibition by sodium chloride

OBJECTIVES: Firefly luciferase, one of the most extensively studied enzymes, has numerous applications. However, luciferase activity is inhibited by sodium chloride. This study was aimed at obtaining mutant luciferase enzymes resistant to the sodium chloride inhibition.

RESULTS: We first obtained two mutant luciferase enzymes whose inhibition were alleviated and determined the mutations to be Val288Ile and Glu488Val. Under medical dialysis condition (140 mM sodium chloride), the wild type was inhibited to 44% of its original activity level. In contrast, the single mutants, Val288Ile and Glu488Val, retained 67% and 79% of their original activity, respectively. Next, we introduced Val288Ile and Glu488Val mutations into wild-type luciferase to create a double mutant using site-directed mutagenesis. Notably, the double mutant retained its activity more than 95% of that in the absence of sodium chloride.

CONCLUSIONS: The mutant luciferase, named luciferase CR, was found to retain its activity in various concentrations of sodium chloride. The luciferase CR may be extensively useful in any bioassay which includes firefly luciferase and is employed in the presence of sodium chloride.

Medienart:

E-Artikel

Erscheinungsjahr:

2021

Erschienen:

2021

Enthalten in:

Zur Gesamtaufnahme - volume:43

Enthalten in:

Biotechnology letters - 43(2021), 8 vom: 04. Aug., Seite 1585-1594

Sprache:

Englisch

Beteiligte Personen:

Yawata, Satoshi [VerfasserIn]
Noda, Kenichi [VerfasserIn]
Shimomura, Ai [VerfasserIn]
Kuroda, Akio [VerfasserIn]

Links:

Volltext

Themen:

451W47IQ8X
Alleviation
EC 1.13.12.7
Journal Article
Luciferase
Luciferases, Firefly
Mutant Proteins
Mutation
Salt inhibition
Sodium Chloride

Anmerkungen:

Date Completed 26.11.2021

Date Revised 26.11.2021

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1007/s10529-021-03109-x

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM324980353