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Use of glucose starvation to limit growth and induce protein production in Escherichia coli
Author(s) -
Tunner Joseph R.,
Robertson Channing R.,
Schippa Serena,
Matin Abdul
Publication year - 1992
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.260400211
Subject(s) - chemostat , escherichia coli , beta galactosidase , lac operon , biology , biochemistry , recombinant dna , operon , starvation , chemistry , bacteria , gene , endocrinology , genetics
The use of glucose starvation to uncouple the production of recombinant β‐galactosidase from cell growth in Escherichia coli was investigated. A lacZ operon fusion to the carbon starvation‐inducible cst ‐ 1 locus was used to control β‐galactosidase synthesis. β‐Galactosidase induction was observed only under aerobic starvation conditions, and its expression continued for 6 h following the onset of glucose starvation. The cessation of β‐galactosidase expression closely correlated with the exhaustion of acetate, an overflow metabolite of glucose, from the culture medium. Our results suggest the primary role of acetate in cst ‐ 1 ‐controlled protein expression is that of an energy source. Using this information, we metered acetate to a glucose‐starved culture and produced a metabolically sluggish state, where growth was limited to a low linear rate and production of recombiant β‐galactosidase occurred continuously throughout the experiment. The cst ‐ 1 controlled β‐galactosidase synthesis was also induced at low dilution rates in a glucose‐limited chemostat, suggesting possible applications to high‐density cell systems such as glucose‐limited recycle reactors. This work demonstrates that by using an appropriate promoter system and nutrient limitation, growth can be restrained while recombinant protein production is induced and maintained.