Heterologous protein production in Escherichia coli biofilms: A non-conventional form of high cell density cultivation. (June 2017)

• Record Type: Journal Article
• Authors: Gomes, L.C.; Mergulhão, F.J.
• Is Part Of: Process biochemistry. Volume 57(2017)
• Page Start: 1
• Page End: 8
• Publication Date: 2017-06
• Subjects: Escherichia coli -- Heterologous protein expression -- High cell density culture -- Biofilm reactor; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2017.03.018
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Specific eGFP production in biofilm was 30 fold higher than in planktonic cells. eGFP-expressing cells had enhanced biofilm formation compared to control cells. Continuous biofilm cultures may be considered as a non-conventional form of HCDC. Abstract: Escherichia coli is one of the favourite hosts for recombinant protein production and is recognized as an excellent model for biofilm studies. High cell density cultures (HCDC) of this bacterium enable attractive volumetric production yields and cells growing in biofilms share some of the challenges of conventional high cell density planktonic cultures. This work assesses the production potential of E. coli JM109(DE3) biofilm cells expressing a model protein, the enhanced green fluorescent protein (eGFP), from a recombinant plasmid. A control strain harbouring the same plasmid backbone but lacking the eGFP gene was used to assess the impact of heterologous protein production on biofilm formation. Results show that specific eGFP production from biofilm cells was about 30 fold higher than in planktonic state. Moreover, eGFP-expressing cells had enhanced biofilm formation compared to control cells. Volumetric production values were 2 fold higher than those previously reported with the same protein and are within the range of what can be obtained by conventional HCDC in the production of soluble proteins. Although the cellular density that was obtained was lower than in conventional HCDC (0.5 fold), this novel system reached good production values which are likely to be improved after optimization of culture conditions.