Abstract
Recombinant protein production in bacterial cells is commonly performed using planktonic cultures. However, the natural state for many bacteria is living in communities attached to surfaces forming biofilms. In this work, a flow cell system was used to compare the production of a model recombinant protein (enhanced green fluorescent protein, eGFP) between planktonic and biofilm cells. The fluorometric analysis revealed that when the system was in steady state, the average specific eGFP production from Escherichia coli biofilm cells was 10-fold higher than in planktonic cells. Additionally, epifluorescence microscopy was used to determine the percentage of eGFP-expressing cells in both planktonic and biofilm populations. In steady state, the percentage of planktonic-expressing cells oscillated around 5%, whereas for biofilms eGFP-expressing cells represented on average 21% of the total cell population. Therefore, the combination of fluorometric and microscopy data allowed us to conclude that E. coli biofilm cells can have a higher recombinant protein production capacity when compared to their planktonic counterparts.
Highlights
Recombinant proteins are currently used in different biotechnological industries and are produced in large amounts in bioreactors [1]
Our research group has previously shown that the presence of the non-conjugative plasmid pET28A in E. coli JM109(DE3) cells increased biofilm formation when compared to a non-transformed strain [18]
We found that the specific eGFP production was lower in planktonic cells and this may have contributed to the higher viability of these cells when compared to biofilm cells
Summary
Recombinant proteins are currently used in different biotechnological industries and are produced in large amounts in bioreactors [1]. Biofilms are communities of surface-attached microorganisms encased in a self-produced extracellular matrix [7] Such biological organization provides distinct characteristics to bacteria compared to their planktonic counterparts [8]. The expression of recombinant proteins in E. coli is commonly accomplished by inserting the gene of interest into a multicopy plasmid [10] that imposes a metabolic burden on the host cell [11]. In planktonic cells, this added metabolic burden may decrease cellular growth rates and biomass yields [11], when the plasmid vector is used for the direct production of a recombinant protein [12]. Our research group has previously shown that the presence of the non-conjugative plasmid pET28A in E. coli JM109(DE3) cells increased biofilm formation when compared to a non-transformed strain [18]
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