{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,23]],"date-time":"2026-02-23T11:51:30Z","timestamp":1771847490895,"version":"3.50.1"},"reference-count":51,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2021,9,17]],"date-time":"2021-09-17T00:00:00Z","timestamp":1631836800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FCT\/MCTES (PIDDAC)","award":["Base Funding - UIDB\/00511\/2020 of the Laboratory for Process Engineering, Environment, Bio-technology and Energy - LEPABE"],"award-info":[{"award-number":["Base Funding - UIDB\/00511\/2020 of the Laboratory for Process Engineering, Environment, Bio-technology and Energy - LEPABE"]}]},{"name":"FEDER funds through COMPETE2020 - Programa Operacional Competitividade e Internacionaliza\u00e7\u00e3o (POCI) and by national funds (PIDDAC) through FCT\/MCTES","award":["PTDC\/BII-BIO\/29589\/2017 - POCI-01-0145-FEDER-029589"],"award-info":[{"award-number":["PTDC\/BII-BIO\/29589\/2017 - POCI-01-0145-FEDER-029589"]}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/141614\/2018"],"award-info":[{"award-number":["SFRH\/BD\/141614\/2018"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["CEECIND\/01700\/2017"],"award-info":[{"award-number":["CEECIND\/01700\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Applied Sciences"],"abstract":"<jats:p>In the present study, the effects of different nutrient media on the development of Escherichia coli biofilms and the production of a heterologous protein were examined. E. coli JM109(DE3) cells transformed with pFM23 plasmid carrying the gene for enhanced green fluorescent protein (eGFP) expression were used. Cells were grown in two different culture media, Lysogenic Broth (LB) and M9ZB, in a flow cell system for 10 days. Epifluorescence microscopy, fluorimetry, and a high-performance liquid chromatography (HPLC) method based on hydrophobic interaction chromatography (HIC) were used to assess bacterial growth, plasmid copy number (PCN), and eGFP production in both planktonic and biofilm cells. The results showed that biofilm development was favored in M9ZB medium when compared with LB. However, the number of eGFP-expressing cells was higher in LB for both planktonic and sessile states (two-fold and seven-fold, respectively). In addition, the PCN in biofilm cells was slightly higher when using LB medium (on average, 29 plasmids per cell versus 20 plasmids per cell in M9ZB), and higher plasmid stability was observed in biofilms formed in LB compared to their planktonic counterparts. Hence, E. coli biofilms grown in LB enhanced both plasmid stability and capacity to produce the model heterologous protein when compared to M9ZB.<\/jats:p>","DOI":"10.3390\/app11188667","type":"journal-article","created":{"date-parts":[[2021,9,17]],"date-time":"2021-09-17T21:23:29Z","timestamp":1631913809000},"page":"8667","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["The Influence of Nutrient Medium Composition on Escherichia coli Biofilm Development and Heterologous Protein Expression"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6907-4548","authenticated-orcid":false,"given":"Alexandra","family":"Soares","sequence":"first","affiliation":[{"name":"LEPABE\u2014Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8992-1097","authenticated-orcid":false,"given":"Luciana C.","family":"Gomes","sequence":"additional","affiliation":[{"name":"LEPABE\u2014Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4840-6299","authenticated-orcid":false,"given":"Gabriel A.","family":"Monteiro","sequence":"additional","affiliation":[{"name":"iBB\u2014Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior T\u00e9cnico, Universidade de Lisboa, 1049-001 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5233-1037","authenticated-orcid":false,"given":"Filipe J.","family":"Mergulh\u00e3o","sequence":"additional","affiliation":[{"name":"LEPABE\u2014Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1016\/j.biotechadv.2009.01.008","article-title":"Production of recombinant proteins by microbes and higher organisms","volume":"27","author":"Demain","year":"2009","journal-title":"Biotechnol. 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