Influence of Surface Material and Nutrient Conditions on Green Fluorescent Protein Production in Escherichia coli Biofilms
Source abstract
Abstract
Escherichia coli biofilms have been investigated as a platform for producing recombinant proteins. This study aimed to assess the effect of different surface materials and culture media on E. coli biofilm formation and enhanced Green Fluorescent Protein (eGFP) production. Three culture media with different carbon and nitrogen sources (Lysogeny broth, Terrific broth, and M9ZB broth) were tested in combination with three materials with distinct surface properties (stainless steel, polyvinyl chloride, and silicone rubber). Biofilm formation, specific eGFP production, and plasmid copy number were monitored in microtiter plates for 9 days. Microscopy and culturability results indicated that biofilm formation was highest in Terrific broth, regardless of the surface material. Additionally, polyvinyl chloride surfaces exposed to Terrific broth provided the most advantageous conditions for achieving the highest specific eGFP production and plasmid maintenance in biofilms. These findings are relevant for establishing operational conditions for producing recombinant proteins and other high-value-added compounds on larger-scale biofilm platforms.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Bacterial biofilms and quorum sensing · Bacterial Genetics and Biotechnology · Bacteriophages and microbial interactions
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
LEPABE—Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal
AliCE—Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal
iBB—Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, University of Lisbon, 1049-001 Lisboa, Portugal
Funding
FCT/MCTES (PIDDAC) · FEDER funds through COMPETE2020-Programa Operacional Competitividade e Internacionalização (POCI) · national funds (PIDDAC) through FCT/MCTES · Portuguese Foundation for Science and Technology (FCT)
References (74)
Show source-provided references
1. Expression in Escherichia coli of chemically synthesized genes for human insulin10.1073/pnas.76.1.106 ↗
2. Recombinant pharmaceuticals from microbial cells: A 2015 update10.1186/s12934-016-0437-3 ↗
3. Puetz, J., and Wurm, F.M. (2019). Recombinant Proteins for Industrial versus Pharmaceutical Purposes: A Review of Process and Pricing. Processes, 7.10.3390/pr7080476 ↗
4. Recombinant protein production in bacterial hosts10.1016/j.drudis.2013.11.008 ↗
5. Rosano, G.L., and Ceccarelli, E.A. (2014). Recombinant protein expression in Escherichia coli: Advances and challenges. Front. Microbiol., 5.10.3389/fmicb.2014.00172 ↗
6. Strategies for achieving high-level expression of genes in Escherichia coli10.1128/mr.60.3.512-538.1996 ↗
7. Production of recombinant proteins by microbes and higher organisms10.1016/j.biotechadv.2009.01.008 ↗
8. Bacterial expression systems for recombinant protein production: E. coli and beyond10.1016/j.biotechadv.2011.09.013 ↗
9. New tools for recombinant protein production in Escherichia coli: A 5-year update10.1002/pro.3668 ↗
10. A concise guide to choosing suitable gene expression systems for recombinant protein production10.1016/j.xpro.2023.102572 ↗
11. Recombinant protein secretion in Escherichia coli10.1016/j.biotechadv.2004.11.003 ↗
12. Strain engineering for improved expression of recombinant proteins in bacteria10.1186/1475-2859-10-32 ↗
13. Efficient genetic approaches for improvement of plasmid based expression of recombinant protein in Escherichia coli: A review10.1016/j.procbio.2017.01.026 ↗
14. Eskandari, A., Nezhad, N.G., Leow, T.C., Rahman, M.B.A., and Oslan, S.N. (2024). Essential factors, advanced strategies, challenges, and approaches involved for efficient expression of recombinant proteins in Escherichia coli. Arch. Microbiol., 206.10.1007/s00203-024-03871-2 ↗
15. Recombinant protein expression in biofilms10.3934/microbiol.2019.3.232 ↗
16. Advanced genetic strategies for recombinant protein expression in Escherichia coli10.1016/j.jbiotec.2004.08.004 ↗
17. Heterologous protein production in Escherichia coli biofilms: A non-conventional form of high cell density cultivation10.1016/j.procbio.2017.03.018 ↗
18. Transcriptome profiles for high-cell-density recombinant and wild-type Escherichia coli10.1002/bit.20340 ↗
19. Stress induced by recombinant protein production in Escherichia coli
20. Effects of medium carbon-to-nitrogen ratio on biofilm formation and plasmid stability10.1002/bit.260440310 ↗
21. Biofilms as living catalysts in continuous chemical syntheses10.1016/j.tibtech.2012.05.003 ↗
22. Microbial biofilms: A concept for industrial catalysis?10.1016/j.tibtech.2009.08.001 ↗
23. Carvalho, F.M., Azevedo, A., Ferreira, M.M., Mergulhao, F.J.M., and Gomes, L.C. (2022). Advances on Bacterial and Fungal Biofilms for the Production of Added-Value Compounds. Biology, 11.10.3390/biology11081126 ↗
24. Soares, A., Gomes, L., Monteiro, G., and Mergulhao, F. (2021). The Influence of Nutrient Medium Composition on Escherichia coli Biofilm Development and Heterologous Protein Expression. Appl. Sci., 11.10.3390/app11188667 ↗
25. Gomes, L., Monteiro, G., and Mergulhao, F. (2020). The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression by Escherichia coli Biofilms. Int. J. Mol. Sci., 21.10.3390/ijms21020576 ↗
26. Soares, A., Gomes, L.C., and Mergulhão, F.J. (2018). Comparing the Recombinant Protein Production Potential of Planktonic and Biofilm Cells. Microorganisms, 6.10.3390/microorganisms6020048 ↗
27. Gomes, L.C., and Mergulhao, F.J. (2017). Effects of antibiotic concentration and nutrient medium composition on Escherichia coli biofilm formation and green fluorescent protein expression. FEMS Microbiol. Lett., 364.10.1093/femsle/fnx042 ↗
28. Temporal variation of recombinant protein expression in Escherichia coli biofilms analysed at single-cell level10.1016/j.procbio.2016.05.016 ↗
29. Flow cell hydrodynamics and their effects on E. coli biofilm formation under different nutrient conditions and turbulent flow10.1080/08927014.2010.535206 ↗
30. Carvalho, F.M., Teixeira-Santos, R., Mergulhao, F.J.M., and Gomes, L.C. (2021). Effect of Lactobacillus plantarum Biofilms on the Adhesion of Escherichia coli to Urinary Tract Devices. Antibiotics, 10.10.3390/antibiotics10080966 ↗
31. Carvalho, F.M., Teixeira-Santos, R., Mergulhao, F.J.M., and Gomes, L.C. (2020). The Use of Probiotics to Fight Biofilms in Medical Devices: A Systematic Review and Meta-Analysis. Microorganisms, 9.10.3390/microorganisms9010027 ↗
32. Escherichia coli adhesion, biofilm development and antibiotic susceptibility on biomedical materials10.1002/jbm.a.35277 ↗
33. Micro- and macro-flow systems to study Escherichia coli adhesion to biomedical materials10.1016/j.ces.2014.12.054 ↗
34. Escherichia coli adhesion to surfaces—A thermodynamic assessment10.1007/s00396-014-3390-x ↗
35. Current and future trends for biofilm reactors for fermentation processes10.3109/07388551.2013.793170 ↗
36. Ercan, D., Pongtharangkul, T., Demirci, A., and Pometto, A.L. (2015). Applications of Biofilm Reactors for Production of Value-Added Products by Microbial Fermentation. Biofims in the Food Environment, John Wiley & Sons, Ltd.10.1002/9781118864036.ch10 ↗
37. Brinson, H., Brinson, L., Brinson, H., and Brinson, L. (2008). Characteristics, Applications and Properties of Polymers. Polymer Engineering Science and Viscoelasticity: An Introduction, Springer.10.1007/978-0-387-73861-1 ↗
38. The Selection of Optimum Media Formulations for Improved Expression of Recombinant Proteins in E. coli
39. Recombinant plasmid retention and expression in bacterial biofilm cultures10.2166/wst.1995.0025 ↗
40. Effects of medium composition and nutrient limitation on loss of the recombinant plasmid pLG669-z and beta-galactosidase expression by Saccharomyces cerevisiae10.1038/sj.jim.2900387 ↗
41. Effects of nutrients on biofilm formation and detachment of a Pseudomonas putida strain isolated from a paper machine10.1016/j.watres.2007.03.041 ↗
42. Overcoming challenges for amplified expression of recombinant proteins using Escherichia coli10.1016/j.pep.2017.11.005 ↗
43. Enhanced bacterial protein expression during auto-induction obtained by alteration of lac repressor dosage and medium composition10.1021/bp070011x ↗
44. Protein production by auto-induction in high density shaking cultures10.1016/j.pep.2005.01.016 ↗
45. Effects of nutritional conditions on plasmid stability and production of tryptophan synthase by a recombinant Escherichia coli
46. Avoiding overfeeding in high cell density fed-batch cultures of E. coli during the production of heterologous proteins10.1016/j.jbiotec.2014.09.002 ↗
47. Muffler, K., and Ulber, R. (2014). Novel Materials for Biofilm Reactors and their Characterization. Productive Biofilms, Springer. Advances in Biochemical Engineering/Biotechnology.10.1007/978-3-319-09695-7 ↗
48. Performance analysis of a pilot-scale membrane aerated biofilm reactor for the treatment of landfill leachate10.1016/j.cej.2015.03.043 ↗
49. Model-based comparative performance analysis of membrane aerated biofilm reactor configurations10.1002/bit.21700 ↗
50. Characterization of planktonic and biofilm cells from two filamentous cyanobacteria using a shotgun proteomic approach10.1080/08927014.2020.1795141 ↗
51. Van Oss, C. (2006). Interfacial Forces in Aqueous Media, CRC Press. [2nd ed.].10.1201/9781420015768 ↗
52. On the Consistency of Surface Free Energy Components as Calculated from Contact Angles of Different Liquids: An Application to the Cholesterol Surface10.1006/jcis.1993.1342 ↗
53. Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors10.1016/0378-1119(85)90120-9 ↗
54. 96-well microtiter plates for biofouling simulation in biomedical settings10.1080/08927014.2014.890713 ↗
55. Flow cells as quasi-ideal systems for biofouling simulation of industrial piping systems10.1080/08927014.2013.821467 ↗
56. Alves, P., Gomes, L.C., Vorobii, M., Rodriguez-Emmenegger, C., and Mergulhao, F.J. (2020). The potential advantages of using a poly(HPMA) brush in urinary catheters: Effects on biofilm cells and architecture. Colloids Surf. B Biointerfaces, 191.10.1016/j.colsurfb.2020.110976 ↗
57. Analysis of factors affecting the periplasmic production of recombinant proteins in Escherichia coli
58. Soares, A., Gomes, L.C., Monteiro, G.A., and Mergulhao, F.J. (2022). Hydrodynamic Effects on Biofilm Development and Recombinant Protein Expression. Microorganisms, 10.10.3390/microorganisms10050931 ↗
59. Quantification of biofilm structures by the novel computer program COMSTAT10.1099/00221287-146-10-2395 ↗
60. Effects of inducer levels on a recombinant bacterial biofilm formation and gene expression10.1007/BF00128622 ↗
61. Plasmid retention and gene expression in suspended and biofilm cultures of recombinant Escherichia coli DH5alpha(pMJR1750)10.1002/bit.260410207 ↗
62. Enhanced high copy number plasmid maintenance and heterologous protein production in an Escherichia coli biofilm10.1002/bit.21240 ↗
63. Chien, H.W., Chen, X.Y., Tsai, W.P., and Lee, M. (2020). Inhibition of biofilm formation by rough shark skin-patterned surfaces. Colloids Surf. B Biointerfaces, 186.10.1016/j.colsurfb.2019.110738 ↗
64. Hydrodynamics and surface properties influence biofilm proliferation10.1016/j.cis.2020.102336 ↗
65. Staphylococcal biofilm growth on smooth and porous titanium coatings for biomedical applications10.1002/jbm.a.34688 ↗
66. Yoda, I., Koseki, H., Tomita, M., Shida, T., Horiuchi, H., Sakoda, H., and Osaki, M. (2014). Effect of surface roughness of biomaterials on Staphylococcus epidermidis adhesion. BMC Microbiol., 14.10.1186/s12866-014-0234-2 ↗
67. Wu, S., Zhang, B., Liu, Y., Suo, X., and Li, H. (2018). Influence of surface topography on bacterial adhesion: A review (Review). Biointerphases, 13.10.1116/1.5054057 ↗
68. Effect of surface chemistry on bacterial adhesion, viability, and morphology10.1002/jbm.a.33178 ↗
69. Enhanced production of recombinant streptokinase in Escherichia coli using fed-batch culture10.1016/j.biortech.2009.04.008 ↗
70. Overproduction of Phytolacca insularis protein in batch and fed-batch culture of recombinant Escherichia coli10.1016/S0032-9592(00)00237-5 ↗
71. Fed-batch operation of recombinant Escherichia coli containing trp promoter with controlled specific growth rate10.1002/bit.260431013 ↗
72. Spontaneous cAMP-dependent derepression of gene expression in stationary phase plays a role in recombinant expression instability10.1016/S0378-1119(98)00020-1 ↗
73. Design of bacterial vector systems for the production of recombinant proteins in Escherichia coli
74. Monmeyran, A., Thomen, P., Jonquiere, H., Sureau, F., Li, C., Plamont, M.A., Douarche, C., Casella, J.F., Gautier, A., and Henry, N. (2018). The inducible chemical-genetic fluorescent marker FAST outperforms classical fluorescent proteins in the quantitative reporting of bacterial biofilm dynamics. Sci. Rep., 8.10.1038/s41598-018-28643-z ↗