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Bioluminescent monitoring of intracellular ATP during fermentation
Author(s) -
Funabashi Hisakage,
Imajo Toshiya,
Kojima Junichiro,
Kobatake Eiry,
Aizawa Masuo
Publication year - 1999
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/(sici)1522-7243(199911/12)14:6<291::aid-bio573>3.0.co;2-i
Subject(s) - luciferase , bioluminescence , plasmid , intracellular , biology , microbiology and biotechnology , escherichia coli , bioreporter , reporter gene , gene expression , biochemistry , chemistry , transfection , gene
The luciferase gene was introduced as a probe into a cell in order to develop a bioluminescent monitoring of intracellular ATP during fermentation. Two plasmids were constructed with two types of promoters. One was pLac–Luc, which had the luciferase gene under the lac promoter to be expressed at a high level. The other was pTet–Luc, which had the luciferase gene under the tetracycline promoter to be stably expressed. A threonine‐overproducing strain of Escherichia coli (No. 29‐4) was transformed with each plasmid. The recombinant E. coli strains were characterized in their growth, threonine production and luciferase expression. The bioluminescence produced intracellularly from expressed luciferase was detected during fermentation in a non‐destructive manner. The bioluminescent intensity reflected both intracellular ATP and luciferase levels, and the results indicate that stable expression of a luciferase reporter is essential for monitoring intracellular ATP. Copyright © 1999 John Wiley & Sons, Ltd.