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 |
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Erscheinungsjahr: |
2021 |
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Erschienen: |
2021 |
Enthalten in: |
Zur Gesamtaufnahme - volume:43 |
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Enthalten in: |
Biotechnology letters - 43(2021), 8 vom: 04. Aug., Seite 1585-1594 |
Sprache: |
Englisch |
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Beteiligte Personen: |
Yawata, Satoshi [VerfasserIn] |
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Links: |
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Themen: |
451W47IQ8X |
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Anmerkungen: |
Date Completed 26.11.2021 Date Revised 26.11.2021 published: Print-Electronic Citation Status MEDLINE |
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doi: |
10.1007/s10529-021-03109-x |
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funding: |
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Förderinstitution / Projekttitel: |
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PPN (Katalog-ID): |
NLM324980353 |
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245 | 1 | 0 | |a Mutant firefly luciferase enzymes resistant to the inhibition by sodium chloride |
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500 | |a published: Print-Electronic | ||
500 | |a Citation Status MEDLINE | ||
520 | |a 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 | ||
520 | |a 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 | ||
520 | |a 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 | ||
650 | 4 | |a Journal Article | |
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700 | 1 | |a Shimomura, Ai |e verfasserin |4 aut | |
700 | 1 | |a Kuroda, Akio |e verfasserin |4 aut | |
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