Details
| Original language | English |
|---|---|
| Pages (from-to) | 315-322 |
| Number of pages | 8 |
| Journal | Biotechnology and bioengineering |
| Volume | 72 |
| Issue number | 3 |
| Publication status | Published - 5 Feb 2001 |
| Externally published | Yes |
Abstract
Aggregation of misfolded proteins can reduce the yield in recombinant protein production. The underlying complex processes are additionally influenced by cellular physiology. Nevertheless, a lumped-parameter model of kinetic competition between folding and aggregation was sufficient to track properly the specific concentration of a human protein produced in E. coli and its partitioning into soluble and insoluble cell fractions. Accurate estimation of the protein-specific parameters required informative experiments, which were designed using the Fisher information matrix. The model was employed to calculate the influence of the specific glucose uptake rate in high-cell-density cultivation of E. coli on accumulation and aggregation of the recombinant protein. Despite its simplicity, the model was flexible and unbiased concerning unidentified mechanisms. Assuming an exponentially decreasing production rate, the irreversible aggregation step was found to follow first-order kinetics, while assuming a constant production rate with simultaneous degradation, the model predicted transient aggregation only. Implications for strain and process development are discussed.
Keywords
- Escherichia coli, Inclusion bodies, Kinetics, Modeling, Protein aggregation, Recombinant protein
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Biotechnology and bioengineering, Vol. 72, No. 3, 05.02.2001, p. 315-322.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Kinetic model of in vivo folding and inculsion body formation in recombinant Escherichia coli
AU - Hoffmann, Frank
AU - Posten, Clemens
AU - Rinas, Ursula
PY - 2001/2/5
Y1 - 2001/2/5
N2 - Aggregation of misfolded proteins can reduce the yield in recombinant protein production. The underlying complex processes are additionally influenced by cellular physiology. Nevertheless, a lumped-parameter model of kinetic competition between folding and aggregation was sufficient to track properly the specific concentration of a human protein produced in E. coli and its partitioning into soluble and insoluble cell fractions. Accurate estimation of the protein-specific parameters required informative experiments, which were designed using the Fisher information matrix. The model was employed to calculate the influence of the specific glucose uptake rate in high-cell-density cultivation of E. coli on accumulation and aggregation of the recombinant protein. Despite its simplicity, the model was flexible and unbiased concerning unidentified mechanisms. Assuming an exponentially decreasing production rate, the irreversible aggregation step was found to follow first-order kinetics, while assuming a constant production rate with simultaneous degradation, the model predicted transient aggregation only. Implications for strain and process development are discussed.
AB - Aggregation of misfolded proteins can reduce the yield in recombinant protein production. The underlying complex processes are additionally influenced by cellular physiology. Nevertheless, a lumped-parameter model of kinetic competition between folding and aggregation was sufficient to track properly the specific concentration of a human protein produced in E. coli and its partitioning into soluble and insoluble cell fractions. Accurate estimation of the protein-specific parameters required informative experiments, which were designed using the Fisher information matrix. The model was employed to calculate the influence of the specific glucose uptake rate in high-cell-density cultivation of E. coli on accumulation and aggregation of the recombinant protein. Despite its simplicity, the model was flexible and unbiased concerning unidentified mechanisms. Assuming an exponentially decreasing production rate, the irreversible aggregation step was found to follow first-order kinetics, while assuming a constant production rate with simultaneous degradation, the model predicted transient aggregation only. Implications for strain and process development are discussed.
KW - Escherichia coli
KW - Inclusion bodies
KW - Kinetics
KW - Modeling
KW - Protein aggregation
KW - Recombinant protein
UR - http://www.scopus.com/inward/record.url?scp=0035809039&partnerID=8YFLogxK
U2 - 10.1002/1097-0290(20010205)72:3<315::AID-BIT8>3.0.CO;2-G
DO - 10.1002/1097-0290(20010205)72:3<315::AID-BIT8>3.0.CO;2-G
M3 - Article
C2 - 11135201
AN - SCOPUS:0035809039
VL - 72
SP - 315
EP - 322
JO - Biotechnology and bioengineering
JF - Biotechnology and bioengineering
SN - 0006-3592
IS - 3
ER -