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Development of a Bacterial Biosensor for Rapid Screening of Yeast p-Coumaric Acid Production
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. Technical University of Denmark.ORCID iD: 0000-0003-4101-855X
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2017 (English)In: ACS Synthetic Biology, E-ISSN 2161-5063, Vol. 6, no 10, p. 1860-1869Article in journal (Refereed) Published
Abstract [en]

Transcription factor-based biosensors are used to identify producer strains, a critical bottleneck in cell factory engineering. Here, we address two challenges with this methodology: transplantation of heterologous transcriptional regulators into new hosts to generate functional biosensors and biosensing of the extracellular product concentration that accurately reflects the effective cell factory production capacity. We describe the effects of different translation initiation rates on the dynamic range of a p-coumaric acid biosensor based on the Bacillus subtilis transcriptional repressor PadR by varying its ribosomal binding site. Furthermore, we demonstrate the functionality of this p-coumaric acid biosensor in Escherichia coli and Corynebacterium glutamicum. Finally, we encapsulate yeast p-coumaric acid-producing cells with E. coli-biosensing cells in picoliter droplets and, in a microfluidic device, rapidly sort droplets containing yeast cells producing high amounts of extracellular p-coumaric acid using the fluorescent E. coli biosensor signal. As additional biosensors become available, such approaches will find broad applications for screening of an extracellular product.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2017. Vol. 6, no 10, p. 1860-1869
Keywords [en]
Para coumaric acid, recombinant DNA, bacterial protein, propionic acid derivative, trans-3-(4'-hydroxyphenyl)-2-propenoic acid, transcription factor, article, bacillus subtilis, bacterium culture, binding site, catalysis, cell population, cell selection, coculture, controlled study, Corynebacterium glutamicum, Escherichia coli, feedback system, fluorescence, fungal cell culture, gene repression, Lactococcus lactis, microbiological examination, microencapsulation, microfluidics, mutagenesis, nonhuman, osmosis, phenotype, priority journal, promoter region, translation initiation, water transport, yeast, yeast cell, genetic procedures, metabolism, procedures, Bacterial Proteins, Biosensing Techniques, Propionates, Transcription Factors
National Category
Cell Biology Other Industrial Biotechnology Biocatalysis and Enzyme Technology
Identifiers
URN: urn:nbn:se:kth:diva-227082DOI: 10.1021/acssynbio.7b00009ISI: 000413715200008PubMedID: 28532147Scopus ID: 2-s2.0-85031721311OAI: oai:DiVA.org:kth-227082DiVA, id: diva2:1206250
Note

QC 20191029

Available from: 2018-05-16 Created: 2018-05-16 Last updated: 2019-10-29Bibliographically approved

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Hammar, PetterJönsson, Håkan

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Khatri, Narendar K.Hammar, PetterJönsson, Håkan
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Proteomics and NanobiotechnologyScience for Life Laboratory, SciLifeLab
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