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2016 ; 15
(1
): 110
Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
The fed-batch principle for the molecular biology lab: controlled nutrient diets
in ready-made media improve production of recombinant proteins in Escherichia
coli
#MMPMID27317421
Krause M
; Neubauer A
; Neubauer P
Microb Cell Fact
2016[Jun]; 15
(1
): 110
PMID27317421
show ga
While the nutrient limited fed-batch technology is the standard of the
cultivation of microorganisms and production of heterologous proteins in
industry, despite its advantages in view of metabolic control and high cell
density growth, shaken batch cultures are still the standard for protein
production and expression screening in molecular biology and biochemistry
laboratories. This is due to the difficulty and expenses to apply a controlled
continuous glucose feed to shaken cultures. New ready-made growth media, e.g. by
biocatalytic release of glucose from a polymer, offer a simple solution for the
application of the fed-batch principle in shaken plate and flask cultures. Their
wider use has shown that the controlled diet not only provides a solution to
obtain significantly higher cell yields, but also in many cases folding of the
target protein is improved by the applied lower growth rates; i.e. final
volumetric yields for the active protein can be a multiple of what is obtained in
complex medium cultures. The combination of the conventional optimization
approaches with new and easy applicable growth systems has revolutionized
recombinant protein production in Escherichia coli in view of product yield,
culture robustness as well as significantly increased cell densities. This
technical development establishes the basis for successful miniaturization and
parallelization which is now an important tool for synthetic biology and protein
engineering approaches. This review provides an overview of the recent
developments, results and applications of advanced growth systems which use a
controlled glucose release as substrate supply.