Development of a Bacterial Biosensor for Rapid Screening of Yeast p-Coumaric Acid Production

• 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.

Ämnesord

NATURVETENSKAP  -- Biologi -- Cellbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Cell Biology (hsv//eng)

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Annan industriell bioteknik (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Other Industrial Biotechnology (hsv//eng)

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Biokatalys och enzymteknik (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Biocatalysis and Enzyme Technology (hsv//eng)

Nyckelord

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

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)