Characterisation of a 3-hydroxypropionic acid-inducible system from Pseudomonas putida for orthogonal gene expression control in Escherichia coli and Cupriavidus necator

Scientific Reports
Erik K R HankoNaglis Malys

Abstract

3-hydroxypropionic acid (3-HP) is an important platform chemical used as a precursor for production of added-value compounds such as acrylic acid. Metabolically engineered yeast, Escherichia coli, cyanobacteria and other microorganisms have been developed for the biosynthesis of 3-HP. Attempts to overproduce this compound in recombinant Pseudomonas denitrificans revealed that 3-HP is consumed by this microorganism using the catabolic enzymes encoded by genes hpdH, hbdH and mmsA. 3-HP-inducible systems controlling the expression of these genes have been predicted in proteobacteria and actinobacteria. In this study, we identify and characterise 3-HP-inducible promoters and their corresponding LysR-type transcriptional regulators from Pseudomonas putida KT2440. A newly-developed modular reporter system proved possible to demonstrate that PpMmsR/P mmsA and PpHpdR/P hpdH are orthogonal and highly inducible by 3-HP in E. coli (12.3- and 23.3-fold, respectively) and Cupriavidus necator (51.5- and 516.6-fold, respectively). Bioinformatics and mutagenesis analyses revealed a conserved 40-nucleotide sequence in the hpdH promoter, which plays a key role in HpdR-mediated transcription activation. We investigate the kinetics and dynamics of...Continue Reading

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Citations

Jul 28, 2018·FEMS Microbiology Letters·Seong Gyeong KimGyoo Yeol Jung
Jun 11, 2019·Frontiers in Bioengineering and Biotechnology·Carsten JersIvan Mijakovic
Jul 22, 2018·Applied and Environmental Microbiology·Swathi AlagesanNigel P Minton
Jun 27, 2019·Biotechnology for Biofuels·Christian Arenas-LópezKlaus Winzer
Jul 30, 2019·ACS Synthetic Biology·Larry J MilletJoshua K Michener
Apr 12, 2018·ACS Synthetic Biology·Erik K R HankoNaglis Malys
Jun 28, 2018·ACS Synthetic Biology·Abayomi Oluwanbe JohnsonTuck Seng Wong
Sep 12, 2021·Scientific Reports·Chandran Sathesh-PrabuSung Kuk Lee

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Datasets Mentioned

BETA
KT2440

Methods Mentioned

BETA
biosensors
Fluorescence
electrophoretic mobility shift assay
biosensor
PCR

Software Mentioned

NEBuilder
Hifi
mfold
WebLogo
Clustal Omega
BPROM
NNPP

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