BioVT-2011-13 BibTeX
@ARTICLE{BioVT-2011-13,
AUTHOR = {M. Kunze and Robert Huber and Claudia Gutjahr and Stefan M\"{u}llner and J. B\"{u}chs},
TITLE = {{Predictive tool for recombinant protein production in E. coli shake flask cultures using an on-line monitoring system}},
JOURNAL = {Biotechnology Progress},
YEAR = {2012},
volume = {1},
number = {28},
pages = {103-113},
month = {},
note = {},
abstract = {As E. coli is well defined with respect to its genome and metabolism, it’s a favored host
organism for recombinant protein production. However, many processes for recombinant
protein production run under suboptimal conditions caused by wrong or incomplete
information from an improper screening procedure because appropriate on-line monitoring
systems are still lacking. In this study the Oxygen Transfer Rate, determined on-line in shake
flasks by applying a RAMOS device (Respiration Activity MOnitoring System), was used to
characterize the metabolic state of the recombinant organisms. Sixteen clones of E. coli SCS1
with foreign gene sequences, encoding for different target proteins, were cultivated in an
autoinduction medium, containing glucose, lactose and glycerol, to identify relationships
between respiration activity and target protein production. All sixteen clones showed a
remarkably different respiration activity, biomass and protein formation under induced
conditions. However, the clones could be classified into three distinct types and correlations
could be made between OTR patterns and target protein production. For two of the three types
a decrease of the target protein was observed after the optimal harvest time had passed. The
acquired knowledge was used to modify the autoinduction medium in order to increase the
product yield. Additional 1.5 g/L glucose accelerated the production process for one clone,
shifting the time point of the maximal product yield from 24 to 17 h. For another clone lactose
addition led to higher volumetric product yields, in fact 25 % and 38 % more recombinant
protein for 2 and 6 g/L additional lactose, respectively.},
keywords = {E. coli screening, recombinant protein, autoinduction, oxygen transfer rate, RAMOS on-line monitoring},
}
Martin Kunze, Robert Huber, Claudia Gutjahr, Stefan Müllner, Jochen Büchs:
Predictive tool for recombinant protein production in E. coli shake flask cultures using an on-line monitoring system
Biotechnology Progress, 2012, 1(28), 103-113
Abstract:
As E. coli is well defined with respect to its genome and metabolism, it’s a favored host
organism for recombinant protein production. However, many processes for recombinant
protein production run under suboptimal conditions caused by wrong or incomplete
information from an improper screening procedure because appropriate on-line monitoring
systems are still lacking. In this study the Oxygen Transfer Rate, determined on-line in shake
flasks by applying a RAMOS device (Respiration Activity MOnitoring System), was used to
characterize the metabolic state of the recombinant organisms. Sixteen clones of E. coli SCS1
with foreign gene sequences, encoding for different target proteins, were cultivated in an
autoinduction medium, containing glucose, lactose and glycerol, to identify relationships
between respiration activity and target protein production. All sixteen clones showed a
remarkably different respiration activity, biomass and protein formation under induced
conditions. However, the clones could be classified into three distinct types and correlations
could be made between OTR patterns and target protein production. For two of the three types
a decrease of the target protein was observed after the optimal harvest time had passed. The
acquired knowledge was used to modify the autoinduction medium in order to increase the
product yield. Additional 1.5 g/L glucose accelerated the production process for one clone,
shifting the time point of the maximal product yield from 24 to 17 h. For another clone lactose
addition led to higher volumetric product yields, in fact 25 % and 38 % more recombinant
protein for 2 and 6 g/L additional lactose, respectively.
Keywords:
E. coli screening, recombinant protein, autoinduction, oxygen transfer rate, RAMOS on-line monitoring
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