Soares Alexandra, Gomes Luciana Calheiros, Mergulhão Filipe José
LEPABE-Department of Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
Microorganisms. 2018 May 24;6(2):48. doi: 10.3390/microorganisms6020048.
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 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 biofilm cells can have a higher recombinant protein production capacity when compared to their planktonic counterparts.
在细菌细胞中生产重组蛋白通常采用浮游培养法。然而,许多细菌的自然状态是生活在附着于表面形成生物膜的群落中。在这项研究中,使用流动池系统比较浮游细胞和生物膜细胞中模型重组蛋白(增强型绿色荧光蛋白,eGFP)的产量。荧光分析表明,当系统处于稳态时,生物膜细胞的平均比eGFP产量比浮游细胞高10倍。此外,采用落射荧光显微镜来确定浮游细胞群和生物膜细胞群中表达eGFP的细胞百分比。在稳态下,浮游表达细胞的百分比在5%左右波动,而对于生物膜,表达eGFP的细胞平均占总细胞群的21%。因此,荧光分析和显微镜数据相结合使我们得出结论,与浮游细胞相比,生物膜细胞具有更高的重组蛋白生产能力。