TY - JOUR

T1 - Global proteome response of Escherichia coli BL21 to production of human basic fibroblast growth factor in complex and defined medium

AU - Li, Zhaopeng

AU - Nimtz, Manfred

AU - Rinas, Ursula

N1 - Funding information: Partial financial support from the German Ministry of Education and Research (BMBF) through the FORSYS-Partner program (grant FKZ 0315285) and from the German Research Council (DFG) through the Cluster of Excellence “Rebirth” EXC62 is gratefully acknowledged.

PY - 2017/8

Y1 - 2017/8

N2 - The global proteome response toward recombinant protein production in Escherichia coli BL21 (DE3) grown in complex and defined medium was analyzed. Overproduction of human basic fibroblast growth factor (hFGF-2), a difficult-to-fold protein, led to a reconstruction of the bacterial proteome. For example, heat shock chaperones were highly upregulated, especially when production occurred during fast growth in complex medium. Although heat shock chaperones increased to higher levels in complex medium more hFGF-2 accumulated within inclusion bodies indicating that the capacity to chaperone protein folding was not sufficient for high speed production. In both types of media, cellular proteins from substrate transport systems, central metabolic pathways, and by-product uptake (e.g. acetate) were downregulated. This downregulation was connected to growth inhibition and metabolic perturbations. For example, during production in complex and defined medium acetate reassimilation and glucose uptake, respectively, were severely hampered. Cellular proteins for degradation of less favorable substrates, elimination of reactive oxygen species, and DNA protection were also downregulated in response to hFGF-2 production. The decrease of proteins involved in transport, central metabolic pathways, and general cell protection was more pronounced in the fast producing culture in complex medium than in the slow producing culture in defined medium. In general, production of hFGF-2 seems to interfere with the adaptation process to changing growth conditions, in this case the adaptation from exponential growth to stationary phase.

AB - The global proteome response toward recombinant protein production in Escherichia coli BL21 (DE3) grown in complex and defined medium was analyzed. Overproduction of human basic fibroblast growth factor (hFGF-2), a difficult-to-fold protein, led to a reconstruction of the bacterial proteome. For example, heat shock chaperones were highly upregulated, especially when production occurred during fast growth in complex medium. Although heat shock chaperones increased to higher levels in complex medium more hFGF-2 accumulated within inclusion bodies indicating that the capacity to chaperone protein folding was not sufficient for high speed production. In both types of media, cellular proteins from substrate transport systems, central metabolic pathways, and by-product uptake (e.g. acetate) were downregulated. This downregulation was connected to growth inhibition and metabolic perturbations. For example, during production in complex and defined medium acetate reassimilation and glucose uptake, respectively, were severely hampered. Cellular proteins for degradation of less favorable substrates, elimination of reactive oxygen species, and DNA protection were also downregulated in response to hFGF-2 production. The decrease of proteins involved in transport, central metabolic pathways, and general cell protection was more pronounced in the fast producing culture in complex medium than in the slow producing culture in defined medium. In general, production of hFGF-2 seems to interfere with the adaptation process to changing growth conditions, in this case the adaptation from exponential growth to stationary phase.

KW - Complex medium

KW - Defined medium

KW - Metabolic burden

KW - Stress response

KW - Two-dimensional gel electrophoresis

UR - http://www.scopus.com/inward/record.url?scp=85021333471&partnerID=8YFLogxK

U2 - 10.1002/elsc.201700036

DO - 10.1002/elsc.201700036

M3 - Article

AN - SCOPUS:85021333471

VL - 17

SP - 881

EP - 891

JO - Engineering in Life Sciences

JF - Engineering in Life Sciences

SN - 1618-0240

IS - 8

ER -