Application of a fed-batch bioprocess for the heterologous production of hSCOMT in Escherichia coli.
In this paper, a fed-batch cultivation process in recombinant Escherichia coli BL21(DE3) bacteria, for the production of human soluble catechol-O-methyltransferase (hSCOMT), is presented. For the first time, a straightforward model is applied in a recombinant hSCOMT expression system and distinguishes an initial cell growth phase from a protein production phase upon induction. Specifically, the kinetic model predicts biomass, substrate, and product concentrations in the culture over time and was identified from a series of fed-batch experiments designed by testing several feed profiles. The main advantage of this model is that its parameters can be identified more reliably from distinct fed-batch strategies, such as glycerol pulses and exponential followed by constant substrate additions. Interestingly, with the limited amount of data available, the proposed model accomplishes satisfactorily the experimental results obtained for the three state variables, and no exhaustive process knowledge is required. The comparison of the measurement data obtained in a validation experiment with the model predictions showed the great extrapolation capability of the model presented, which could provide new complementary information for the COMT production system.
Passrinha
L A
LA
CICS - Centro de Investigagao em Ciencias da Saude, Universidade da Beira Interior 6201-001 Covilha, Portugal. lpassarinha@fcsaude.ubi.pt
Bonifacio
M J
MJ
Queiroz
J A
JA
D019149:Bioreactors; D002138:Calibration; D002394:Catechol O-Methyltransferase / Q000235:genetics* / Q000378:metabolism; D002455:Cell Division; D003470:Culture Media; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008954:Models, Biological; D011994:Recombinant Proteins / Q000378:metabolism; D013696:Temperature; D014162:Transfection
D003470:Culture Media; D011994:Recombinant Proteins; D002394:Catechol O-Methyltransferase