Synthetic Biology Tools for the Membrane – Targeted Localisation and Elucidation of Protein Interactions

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedings – Annual report year: 2014Researchpeer-review

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To meet the need for new, green production scenarios, development of biological cell factories is becoming increasingly important. In order for cell factories to compete with traditional production means, it is essential to expand the available toolbox. We are developing tools for the E. coli cell factory, focusing on the membranes. Surface displaying enzymes may allow for development of whole-­‐cell catalysts, but a recurring issue with the technique is to know that a protein is indeed surface displayed, and not detected due to lysis of cells or secretion. We are developing a fluorescence-­‐based method for easy detection of surface display using GFP-­‐nanobodies (1). We fused a GFP-­‐nanobody to the E. coli outer membrane anchor LppOmpA, resulting in surface display of the nanobody. Addition of free GFP to the whole cells resulted in binding of GFP to the cells via the nanobody, as shown by fluorescence measurements, in-­‐gel fluorescence and microscopy (fig. 1). We are now further expanding the toolbox for use in surface display applications. In addition, to expand our knowledge of protein interactions and protein complex formation in the membrane, we are exploring the use of styrene maleic acid (SMA) for isolation of membrane proteins. SMA is a polymer which spontaneously digs into a lipid membrane and carves out a disc containing protein and native lipids (2). By elucidating protein interactions we will be able to tune and optimise heterologous pathway expression in our E. coli cell factories.
Original languageEnglish
Title of host publicationAbstract Book - DTU Sustain Conference 2014
Number of pages1
Place of PublicationKgs. Lyngby
PublisherTechnical University of Denmark
Publication date2014
Publication statusPublished - 2014
EventDTU Sustain Conference 2014 - Technical University of Denmark, Lyngby, Denmark
Duration: 17 Dec 201417 Dec 2014
http://www.sustain.dtu.dk/

Conference

ConferenceDTU Sustain Conference 2014
LocationTechnical University of Denmark
CountryDenmark
CityLyngby
Period17/12/201417/12/2014
Internet address

ID: 104280300