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Use of culture fluorescence as a sensor for on‐line discrimination of host and overproducing recombinant Escherichia coli
Author(s) -
Walker Colleen C.,
Dhurjati Prasad
Publication year - 1989
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260330415
Subject(s) - recombinant dna , escherichia coli , biology , plasmid , intracellular , enterobacteriaceae , fluorescence , biochemistry , cell culture , host (biology) , microbiology and biotechnology , gene , genetics , physics , quantum mechanics
The culture fluorescence of two recombinant Escherichia coli strains with high plasmid copy number were studied and compared to both the host and low copy number varieties of the corresponding strains. Culture fluorescence data are related to the concentration of reduced intracellular nicotinamide adenine dinucleotide within a cell, and can therefore be used as a means for detecting changes in metabolic states. Correlation curves relating culture fluorescence to biomass show that the recombinant system maintains a larger pool of intracellular NADH at high plasmid copy numbers than either the host or the recombinant system at low copy numbers. These results demonstrate the ability of a fluorescence probe to detect differences in the metabolic demands made on an over‐producing recombinant organism.